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Notes: Summary Clones of Phryganella acropodia were cultivated under different trophic conditions with bacteria as the food source. The doubling time was estimated to be 3 days. The edibility of four species of fungi, Aspergillus niger, Cunninghamella echinulata, Penicillium echinulatum and Stilbella bulbicola, was tested, but only Penicillium enchinulatum and Stilbella bulbicola were eaten and digested by the amoeba. An ultrastructure examination showed that there are two contractile vacuoles, many dictyosomes, a single nucleus with several nucleoli, and peroxisomes. The pseudopodia are filiform when attached to the substrate but change to lobose when the animal is floating. A thin organic membrane covers the aperture of resting forms.

Notes: Summary The microvasculature of the stroma of human gastric carcinoma was studied by immuno-electron microscopy for factor VIII/von Willebrand factor (vWF) and conventional electron microscopy. In differentiated type (intestinal) gastric carcinoma (9 cases), capillaries were distributed more densely around carcinoma cell nests. vWF was localized in endothelial cells and neighbouring stroma. Ultrastructurally, capillary endothelial cells showed considerable hypertrophic changes with well-developed rough endoplasmic reticulum (rER). vWF was localized in well-developed rER, granules, Weibel-Palade bodies (WPB), in the vascular lumen as clusters, and diffusely deposited in the subendothelium. This indicates that endothelial cells in this group are transformed into a state of active protein production. In undifferentiated type (diffuse) gastric carcinoma (12 cases), capillaries were uniformly distributed and endothelial hypertrophic changes were less remarkable. vWF was localized in WPB, scanty rER and subendothelial matrix. Solid capillary buds were observed in both types; they were composed of a solid strand of endothelial cells without a visible lumen. Our results reveal that the microvasculature in tumour stroma differs significantly according to its histological type.

Notes: Summary Ciliated cells were found in the gastric mucosa in close association with intestinal metaplasia, mainly in the pyloric mucosa, of Japanese patients. The occurrence of ciliated cells is believed to be an acquired phenomenon and is considered to be a type of metaplasia; the term “ciliated metaplasia” is used to describe this phenomenon. Ciliated cells are found in the basal part of the glands among normal-looking mucous cells, mucous neck cells and neuroendocrine cells, but never on the surface or in foveolar epithelium. In ciliated cellcontaining glands, mitoses were noted in the neck region and the ultrastructural features of these cells were identical to those of undifferentiated neck cells. However, cell metaplasia from undifferentiated cells to metaplastic ciliated cells has never been demonstrated previously. The small mucus-granule-containing ciliated cells found in our present study may arise subsequent to division of undifferentiated neck cells into mucous cells with some daughter cells then exhibiting differentiation characteristics specific to ciliated cells. Thus they contain a mixture of both small mucus granules and numerous basal bodies and cilia, at the same time as a transitional form.

Notes: Summary We performed an ultrastructural, morphometric comparison of mitochondria and myofibrils of cardiomyocytes using endomyocardial biopsy specimens in hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM). Biopsies came from the right ventricular side of the interventricular septum in nine patients with HCM, nine with DCM, and nine controls with arrhythmia and/or ST depression. Morphometric analysis was carried out using electron microscopic photographs and an image analyser. Mitochondria were significantly greater in number and smaller in size in HCM than in the control group. In DCM, the size of mitochondria was also significantly smaller than in the control group, although their number was similar to that of the control group. No statistically significant difference was found regarding the size of mitochondria between HCM and DCM. The percentages of both mitochondrial and myofibrillar areas in cytoplasm were smaller in the DCM than the HCM and control groups, though no difference was seen between the latter two. The ratio of mitochondrial area to myofibrillar area was almost the same in each group. These results suggest increased mitochondrial function to match hypertrophic cardiomyocytes in HCM, and decreased mitochondrial function and cardiomyocytic contractility in DCM.

Notes: Summary So-called minute pulmonary chemodectoma is a curious, small lung tumour found mainly in women. The nature and origin of the proliferating cells are still obscure. In the first report on the tumour, the component cells were described as resembling chemoreceptor cells and the tumour was named chemodectoma. However, electron microscopic studies of the tumour have revealed no evidence of neuronal characteristics and have shown a close resemblance to meningothelial cells. In this study, the electron microscopic findings were similar to those previously reported but in one of the two cases, tumour cells were filled with abundant cytofilaments, giving them an occasional dense, patch-like appearance. Immunostaining for myosin and vimentin was positive in all tumour cells, but epithelial membrane antigen staining was not seen. These findings indicate that the tumour might have its origin from muscle cells.

Notes: Summary New ultrastructural findings are reported from two lymphomas of vacuolar signet-ring cell morphology (SR+), one of B cell and one of T cell lineage. When these lymphomas were compared ultrastructurally a difference in the relationship of the endoplasmic reticulum (ER) to the vacuole was noted, although the fine structure of the vacuoles themselves was similar and they were interpreted as giant multivesicular bodies (mvbs). Smooth ER was found near the vacuoles in both cases. Dark mvbs with a complex, reticulate form are emphasised as readily identified but hitherto unreported cell components in these tumours. A further B cell lymphoma of centroblastic/centrocytic type which was SR− was found to be rich in mvbs and may be a transitional form between SR− and SR+ lymphomas. In addition, the occurrence of mvbs has been studied quantitatively in a number of other lymphomas and in B and T lymphocytes in reactive nodes. Although increased numbers of mvbs were found in neoplastic compared with reactive lymphocytes, and in T compared with B cell lymphomas, these differences were not statistically significant. The possible roles of endoplasmic reticulum and mvbs in the generation of SR+ change are discussed.

Notes: Summary Contraction bands usually occur in the intramural working myocardium following post-ischaemic reperfusion. In the subendocardium, however, they are found during ischaemia. Thus, we ascertained the contraction states of Purkinje fibres, transitional cells, subendocardial and intramural parts of the working myocardium during 30 min global ischaemia at 25° C. The effects with and without myocardial protection were compared. At the onset of pure ischaemia contraction bands are completely lacking in all cell types. During pure ischaemia contraction bands are found in all subendocardial cell types but not in the intramural working myocardium. A peak of pathological contraction states is found in the intramural working myocardium at the onset (0 min), in the subendocardial working myocardium at 10 min, in the transitional cells and Purkinje fibres at 30 min of pure ischaemia. Histidine-, tryptophan-, ketoglutarate-enriched (HTK) cardioplegia prevents contraction bands completely at the onset of ischaemia and prevents both contraction bands and pathological contraction states during ischaemia almost completely. Striking differences in the physiological contraction states are seen only in the working myocardium: HTK cardioplegia brings about dominance of relaxation during ischaemia. These findings may be due mainly to the effects of global ischaemia on the one hand and to catecholamines, calcium and oxygen on the other.

Notes: Summary Well-differentiated papillary mesothelioma (WDPM) is considered to be a distinct subtype of peritoneal mesothelioma. Although the WDPM is usually qualified as benign, the natural history of this lesion has not been clearly established. This report relates to two WDPMs which were found incidentally. In a 51-year-old man the WDPM developed over a period of 5 years into a typically malignant diffuse mesothelioma. Thus, although the WDPM morphologically lacks signs of malignancy, it should be regarded as a borderline mesothelioma.

Notes: Summary Patients with olivopontocerebellar atrophy (OPCA) were studied, and cytoplasmic inclusions were observed in some of the remaining neurons of the pontine nuclei, nuclei reticularis tegmenti pontis and arcuate nuclei. The cytoplasmic argyrophilic inclusions were demonstrated by silver impregnation techniques such as Bielschowsky and Bodian staining. With hematoxylin and eosin stain, the inclusions were sharply demarcated and appeared pale. The inclusions were not stained by the following routine histological methods: Klüver-Barrera, phosphotungstic acid hematoxylin, Holzer, periodic acid-Schiff, Mallory azan, alcian blue, nile blue, Masson trichrome, Congo red, thioflavine S, oil red O and Sudan black B stains. Immunohistochemistry with anti-ubiquitin antiserum showed that these inclusions were ubiquitinated. However, the inclusions did not react with any of the following antibodies (Abs) or antisera: anti-phosphorylated neurofilament (NF) Ab, anti-nonphosphorylated NF Abs (160 and 200 kDa), anti-paired helical filament antiserum, anti-tau antiserum, anti-tubulin Abs (alpha and beta), anti-microtubule-associated proteins antiserum, anti-glial fibrillary acidic protein antiserum, anti-vimentin Ab, anti-desmin Ab, anti-cytokeratin Abs (low and high molecular weights), anti-actin antiserum, anti-skeletal myosin antiserum and anti-myelin basic protein Ab. Ultrastructurally, the inclusion bodies noted in OPCA were composed primarily of fibrils having a width ranging from about 24 to 40 nm, which were entirely coated with osmiophilic granular material along their whole length. They were occasionally intermingled with a few filaments about 10 nm in width. Electron microscopical examination on silver-impregnated specimens revealed that each granule-coated fibril had a great affinity for silver particles. In elucidating the pathogenesis of OPCA, it was considered to be an important neuropathological finding that some of the remaining pontine neurons affected by OPCA developed characteristic cytoplasmic argyrophilic inclusions.

Notes: Summary The general gill morphology of Oreochromis alcalicus grahami, a teleost adapted to high salinity and hyperosmosis, is basically similar to that of other teleostean fish. The species has four pairs of gill arches, all of which have well developed filaments. Each of the arches (holobranchs) has two rows of filaments (hemibranchs). Bilaterally situated secondary lamellae branch from the central axis of the filaments. The lamellae reach their maximum size at the middle of the filament, gradually decrease in size and eventually disappear towards the tip of the filament, which is bare. The leading edge of the gill filament and the immediate interlamellar space is covered by a stratified epithelium consisting of pavement cells, mucous cells, chloride cells and undifferentiated cells. The surface of these cells is made up of concentric microridges. The chloride cells were found only on the primary epithelium (filamental epithelium) and very rarely on the secondary epithelium (lamellar epithelium). Two types of chloride cells were observed in the gills of Oreochromis. The superficial chloride cells have fewer mitochondria concentrated towards the basal aspect of the cell, and a network of tubules towards the apical surface and are less electron dense. These cells intercommunicate with the water through an apical pore. The deep chloride cells have numerous diffuse mitochondria intercalated between a fine profuse tubular network and are more electron dense. These cells are covered by one or more layers of pavement cells and thus do not have access to the external surface. After gradual dilution of the lake water in which the fish were kept, both types of chloride cells remained topographically and ultrastructurally distinct. However, in both kinds of cell the mitochondria decreased in number and size. Initially there was an increase in the diameter and the degree of interdigitation of the tubules followed by a gradual decrease. An increase in the quantity of rough endoplasmic reticulum, particularly at the perinuclear region of the cell, was noted. The morphometric analysis of the branchial system indicated that the gills of Oreochromis are well adapted for gas exchange by having numerous and relatively long gill filaments with a high lamellar density. These features provide a large surface for gas exchange which, when coupled with the notably thin water-blood barrier of an average thickness of only 0.83 μm, would facilitate efficient absorption of oxygen by the gills. Oreochromis alcalicus was observed to be incapable of adapting to freshwater. This may have been due to the progressive degeneration of the chloride cells. Also the arrangement of the chloride cells as a continuum from the central venous compartments to the free epithelial surface is a structural feature which may not be amenable to radical functional changes.

Notes: Summary Physiological cell death is a widespread phenomenon in the development of both vertebrates and invertebrates. This review concentrates on an aspect of developmental cell death that has tended to be neglected, the manner in which the cells are dismantled. It is emphasized that the dying cells may adopt one of at least three different morphological types: “apoptotic”, “autophagic”, and “non-lysosomal vesiculate”. These probably reflect a corresponding multiplicity of intracellular events. In particular, the destruction of the cytoplasm in these three types appears to be achieved primarily by heterophagy, by autophagy and by non-lysosomal degradation, respectively. The various mechanisms underlying both nuclear and cytoplasmic destruction are reviewed in detail. The multiplicity of destructive mechanisms needs to be born in mind in studies of other aspects of cell death such as the signals which trigger it, since different signals probably trigger different types of cell death.

Notes: Summary We carried out a histological examination of the extraocular muscles (EOMs) in a case of myoclonus epilepsy associated with ragged-red fibers (MERRF) and two cases of mitochondrial myopathy, encephalopathy, laetic acidosis, and stroke-like episodes (MELAS), which did not manifest external ophthalmoplegia clinically. By light microscopy, many granular and vesicular fibers were seen associated with endomysial fibrosis. Electron microscopy revealed that the fibers showed prominent accumulation of abnormal mitochondria, extensive loss of myofibrils, proliferation of free sarcoplasmic reticulum and an increased amount of lipid vacuoles. These changes were more pronounced in MELAS than in MERRF. Hirano bodies were often seen in the subsarcolemmal area of muscle fibers and also in the intramuscular myelinated nerve fibers and axon terminals. These findings suggest the presence of mitochondrial myopathy of the EOMs in cases of MELAS and MERRF.

Notes: Summary Ultrastructural damage leading to delayed neuronal death was investigated in the mid-CA1 region of the hippocampus from the stratum (str.) moleculare to oriens after transient bilateral forebrain ischemia in Mongolian gerbils. After ischemia for 5 min without recirculation, mild swelling of the peripheral part of the apical and basal dendrites was already apparent in the str. moleculare and str. oriens. Mitochondria in the dendrites were also swollen in the same area. During recirculation for 12 h to 3 days, swelling of the dendritic cytoplasm persisted with formation of microvacuoles, but swelling of mitochondria receded. Microvacuolation and loss of microtubules were also observed in the proximal part of the dendrites during this period, and swelling and disruption of internal cristae were observed in mitochondria after recirculation for 3 days. The dendrites became severely degenerated after recirculation for 4 days. In the pyramidal cell bodies, no abnormality was observed at the end of ischemia for 5 min, but disaggregation of polyribosomes and swelling of the endoplasmic reticulum were observed 12 h after recirculation. Proliferation of the endoplasmic reticulum in parallel arrays occurred after recirculation for 1 day and persisted. Severe degeneration of the pyramidal cell bodies was obvious after recirculation for 4 days. The findings observed in the present investigation suggested that the neuronal structure most vulnerable to ischemia was the peripheral part of the dendrites and postischemic neuronal damage occurred early in this part of the dendrites.

Notes: Summary Three cases of spontaneous olfactory neuroblastoma (ONB) in domestic cats were morphologically and immunocytochemically characterized. Diagnostic light microscopic features included Flexner and Homer-Wright rosettes, while ultrastructurally the cells had neuritic processes, intracellular intermediate filaments, and intercellular junctions. Immunocytochemically, the tumors stained positively for neuron-specific enolase, cytokeratins, and S-100 protein antigens. In each case, a key finding was the identification of numerous mature type C retroviral particles within the tumors. In one case, budding of viral particles from the plasmalemma of tumor cells suggested the source of mature particles. This cat and one other were tested, and both were serologically positive for feline leukemia virus (FeLV). The virus in the tumors was identified as FeLV by polymerase chain reaction and immunocytochemistry. No other neoplasms were found in any of the cats, nor was there similar evidence of active viral infection in other non-tumor tissues, including the brain. Although the relationship between FeLV infection and ONB is uncertain, our findings indicate that FeLV should be investigated as an etiologic agent of ONB.

Notes: Summary Large intracytoplasmic inclusions arising from the endoplasmic reticulum and nuclear envelope were found in the muscle fibers of a 32-year-old individual with a life-long, chronic and progressive sensory-motor neuropathy. The morphological features of the inclusions were similar to that occasionally reported in the striated muscles in several unrelated conditions as well as to “hyaline” or “colloid” inclusions commonly seen in motor neurons of the brain stem and spinal cord. The chemical nature of the inclusions is not known. Their occurrence in the muscle fibers is probably secondary to chronic denervation.

Notes: Summary Intracellular inclusions with staining properties of amyloid are very common in the aging choroid plexus epithelial cells. In many ways these inclusions show similarities with the neurofibrillary tangles, found in cerebral cortical neurons in patients with Alzheimer's dementia. We have now designed a purification method for choroid plexus amyloid and performed a transmission and scanning electron miscroscopic study. This shows that one form of choroid plexus inclusions, the Biondi ring, is a homogeneous globule covered with a thin layer of amyloid fibrils. Partial immunochemical characterization of the choroid plexus amyloid reveals that it is different from the neurofibrillary tangles although there are similarities.

Notes: Summary Scanning electron microscopic observations of connective tissue cells show a new aspect of the nature of fibroblasts, and the subsequent broad survey of references makes clear that fibroblasts of many tissues have various features which are regarded as atypical of fibroblasts, and at the same time that various connective tissue cells in different organs have features typical of fibroblasts. Both morphological and functional features of fibroblasts are more or less common to those of fibroblast-like cells, and differences among these cells are quantitative rather than qualitative. Therefore, it is almost impossible to set clear-cut criteria for distinguishing genuine fibroblasts from a large population of fibroblast-like cells. The majority of cells sharing features of fibroblasts, if not all, seem to belong to the same population of cells. They are probably adapted to special functional needs in their own micro-environment that are peculiar to local or pathological or experimental conditions. It is proposed to categorize these cells into subtypes depending on their main functions: 1, fibrogenesis; 2, tissue skeleton or barrier; 3, intercellular communication system; 4, gentle contractile machinery; 5, endocrine activity; and 6, vitamin A-storing. Re-evaluation of fibroblasts and fibroblast-like cells is required to facilitate their better understanding.

Notes: Summary Blood samples were taken weekly from seventeen mature blue fox vixens (average age five years), from late anoestrus until pro-oestrus, and then taken daily. The vixens were sacrificed at various stages of oestrus, and oocytes were collected from ovarian follicles by aspiration, and/or from oviducts by flushing. The structural features of oocyte maturation were related to the time of the luteinizing hormone (LH) peak. On days 1–2 after the LH peak the oocyte nucleus migrated from a central to a peripheral position in the ooplasm and assumed a flattened appearance. The cumulus investment expanded simultaneously and ovulation took place around day 2. On days 2–3 the oocyte nuclear envelope broke down, the nucleoli disappeared, the metaphase of the first meiotic division was reached, the Golgi complexes decreased in size, the perivitelline space enlarged, and all junctional contact between cumulus cell projections and oocyte was disrupted. On days 3–5 the first polar body was extruded, the metaphase of the second meiotic division was reached, and the cumulus cells degenerated. On day 5 the release of cortical granule content was occasionally seen, and from day 6 the oocytes showed signs of degeneration. In a few animals deviant oocyte maturation was noticed.

Notes: Summary In five species of lungless salamanders, family Plethodontidae, which all show highly developed visual abilities, the ultrastructure of the optic nerve was investigated and the total number of retinal ganglion cell axons, the percentage of myelinated axons, and the volume densities of glia and axons were determined. More than 80% of all axons were smaller than 0.4 μm and only 2–3% were larger than 0.8 μm. In individual nerves the degree of myelination varied between 1 and 9% which is in the range reported for other amphibian species. The miniaturized and highly paedomorphic species Batrachoseps attenuatus was an exception because only very few or even no myelinated axons were present in the nerve, which is unique among gnathostome vertebrates. The five investigated species had total numbers of axons ranging from 26000 in Batrachoseps attenuatus to about 50000 in Plethodon jordani. These numbers are the lowest found among vertebrates with an elaborated visual system. The amount of glial material in the optic nerve varied between 25 and 50%, with larger nerves possessing more glia than smaller ones. Ultrastructural analysis revealed that the optic nerve of each species contained both astrocytes and oligodendrocytes, although often in immature form. In Batrachoseps attenuatus the glia showed features of both astrocytes and oligodendrocytes which reflect an undifferentiated state.

Notes: Summary The ultrastructural morphology of developing chorionic villi in the marmoset monkey (Callithrix jacchus) placenta was studied in pregnant monkeys at known time intervals after ovulation. In samples obtained at 45 days after ovulation the mesoderm, which consists of primitive foetal blood vessels, is seen to extend down into cytotrophoblast columns. Syncytiotrophoblast completely surrounds maternal blood vessels and both basal laminae and endothelial cells of maternal origin show signs of disorganisation and degradation. Syncytiotrophoblast is first observed to breach the maternal circulation in samples collected from animals at 60 days after ovulation; this results in discrete haemochorial villi randomly distributed throughout the placental bed. Samples obtained at 80 days after ovulation and term placental samples (145 days after ovulation) exhibit tertiary haemochorial villi throughout the placenta, similar to those seen randomly distributed at 60 days after ovulation.

Notes: Summary The early development of anuran posterior lymph hearts studied by light- and electron-microscopy in frog larval stages 25–29 (Gosner 1960) can be subdivided into three phases. In phase I, mesenchymal myogenic cells are found, each possessing a single 9 + 0 cilium and numerous filopode-like processes aggregated near the vena caudalis lateralis, forming up to three metameric organ anlagen arranged like a cuff around the preexisting lymphatics (stages 26/27). In phase II, cell proliferation starts at stage 28 within the lymph heart wall as does the formation of primarily polynuclear myofibres by fusion of several myoblasts. At this stage immature myofibres show a vast sarcoplasm, a poorly developed SR and only few myofibrils with not yet distinguishable A- and I-bands. In phase III, the afferent and efferent valves are formed at the onset of pulsation in stage 29. Contractile myofibres contain large glycogen fields and a considerable amount of myofibrils which frequently branch and show distinct Z-lines, A-, I-, H- and M-bands; 1–3 cilia were found lying within a channel-like cell invagination. The peculiarities of organogenesis and myofibre development are discussed.

Notes: Summary The nature of Rosenthal fibres (RF) was investigated in eight cases each of low-grade astrocytoma and reactive gliosis using immunohistochemical (IH) staining for glial fibrillary acidic protein (GFAP), electron microscopy (EM) and immunoelectron microscopy (IEM) by immunogold labelling technique. By IH under light microscopy (LM), three types of RF were seen, uniformly positive (type I), rim positive (type II) and completely negative (type III). EM showed variation in structural pattern of RF. Some RF contained large amount of glial filaments (GF) intermingled with RF while others with a large amount of electron dense material and less GF. Thus, the presence and amount of GF in RF appear to be responsible for the different types of IH staining under LM. IEM showed that all RF including the ones consisting of entirelh amorphous material possess immunoreactivity for GFAP. It is suggested that RF formation is a two-stage process, staring with excessive accumulation of GF within astrocytic processes followed by their gradual alteration into electron-dense amorphous material under the influence of some unknown metablic or other factors. The quantitative analysis of different types of RF suggests a difference in the rate of formation of RF in neoplastic and reactive conditions.

Notes: Summary A light and electron microscopic study was undertaken to determine pathological changes in cutaneous spinal nerves of rabbits following intradermal inoculation with alphaherpesvirus saimiri (αHVS) isolate KM 322. Infected rabbits were killed at 3, 10, 17, 45 days and 2 years after infection. No abnormalities were seen at 3 days postinoculation. In the nerves of the rabbits killed at 10, 17 and 45 days after infection, axonal (Wallerian-type) degeneration was the main pathological feature. Regeneration, manifested by axonal sprouting, was observed in the nerves of the rabbits killed at 45 days post-inoculation. Neural fibrosis and paucity of unmyelinated axons was the final outcome. The severity of the neural damage not only varied according to the progression of the disease but between nerves taken from the same rabbit. This was probably associated with variation in the numbers of virus particles that had reached the dorsal root ganglion of the dermatome served by a particular nerve. Since αHVS (isolate KM 322) provides a model system for the study of virus latency in dorsal root ganglia, and consequently for the study of varicellazoster infection in man, these findings give further insight into the pathology of herpetic neuropathy.

Notes: Summary Phospholipids in meningiomas were studied by light and electron microscopy, and by high-performance liquid chromatography. They were microscopically demonstrated in six of the ten cases by Sudan III staining after the fixation with potassium dichromate. However, the conventional ultrastructural fixation with glutaraldehyde and osmium tetroxide failed to confirm phospholipids, as most of them were dissolved during dehydration. In contrast, the specimens pretreated with tannic acid before osmication ultrastructurally retained phospholipids which were represented by multilamellar bodies or ribbon-like rings. Both were found in 23 of the 30 cases within the cytoplasm, among the plasma membranes and in the extracellular matrices. The outermost lamella or ribbon showed a direct continuity from the neighbouring plasma membranes of the cytoplasm or the mitochondria. The multilamellar bodies showed an overall distribution, while the ribbon-like rings were predominantly distributed around the psammoma bodies. Precipitation of hydroxyapatite crystals within the ribbon-like rings resulted in matrix minerals of psammoma bodies. Chromatographical analyses of meningiomas disclosed phospholipids including phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl serine, sphingomyelin and phosphatidyl inositol in that order. Psammomatous meningiomas contained a higher percentage of phosphatidyl serine than non-psammomatous tumors. Ultrastructural study of synthetic phospholipids adequately treated with tannic acid showed that the multilamellar bodies were similar to phosphatidyl choline, while the ribbon-like rings were similar to phosphatidyl serine. The role of phospholipids in meningiomas is discussed.

Notes: Summary In this EM study of lateral muscle in Dicentrarchus labrax, we observed that during the larval period, growth of the presumptive red and white muscle layers occurs both by hypertrophy (as fibres already present at hatching complete their maturation) and by production of new fibres in germinal zones specific to the two muscle layers. In the first half of larval life the presumptive white muscle increases in thickness by the addition, superficially, of new fibres derived from a germinal zone of presumptive myoblasts lying beneath the red muscle layer. In the second half of larval life new fibres produced in this same zone form the intermediate (or pink) muscle layer. Dorsoventrally the myotome grows throughout larval life, largely by addition of new fibres from germinal zones at the hypo- and epi-axial extremities. Towards the end of larval life all these germinal zones are becoming exhausted, but another source of fibres arises as satellite cells, associated with large-diameter presumptive white muscle fibres, are activated to produce new fibres. The addition of small, new fibres gives the white muscle its mosaic appearance. Morphometric analysis of fibre diameters in the white muscle confirms that whereas these hyperplastic processes are important during the larval and juvenile periods, when growth is very rapid, they have ceased by the time the adult stage is attained. By contrast, fibre hypertrophy continues through into adult life. The presumptive red muscle consists initially of a monolayer of fibres present only near the lateral line, and during larval life it grows hypo- and epi-axially by addition of fibres derived from myoblasts already present in these areas at hatching. Lying superficially to the presumptive red muscle monolayer there is a near-continuous layer of external cells with a “flattened” profile. During the second half of larval life, differentiation of these external cells into myoblasts provides the source of new fibres which are added to the red muscle layer. This process, which occurs initially in the region around the lateral line and later spreads outwards, is responsible for the increase in thickness of the red muscle.

Notes: Summary The regression of the pronephric kidney of the lamprey, Petromyzon marinus, is described using histochemical and ultrastructural techniques. Regression begins in the third year of larval life, and by the time the animal enters metamorphosis the tubules have all disappeared. The nephrostomes and the renal corpuscle, however, persist for the remainder of the life cycle and undergo little change in the larva. Iron is present within the tubular epithelium prior to the beginning of degeneration, but as degeneration proceeds iron is observed within the tubule lumina. Acid phosphatase is noted within the tubule epithelia prior to degeneration, but as degeneration proceeds acid phosphatase is also observed within the intertubular area. Features of tubular regression include a prominent and highly folded basal lamina, numerous cytoplasmic inclusions, and dense bodies in the epithelia and lumina. The intertubular region is invaded by lymphocytes, granulocytes, plasma cells, and macrophages. The process of pronephric regression possesses many features of the process of apoptosis, which has been noted in the regression of larval organs in other vertebrates.

Notes: Summary The caudal neurosecretory complex (CNc) of poecilids has previously been shown to receive serotonergic inputs. In the present study, immunohistochemical techniques were applied at the light and electron microscopic levels to characterize serotonergic terminals in the neuroendocrine nucleus. A dense plexus of varicose fibers observed in the rostral CNc neuropil was absent in the spinal cords of deafferented fish, indicating that the origin of this input was extranuclear. Ultrastructural study revealed no direct contacts between labeled structures and neuroendocrine cells. Non-synaptic terminals (varicosities) were the predominantly labeled structures in the neuropil. Synaptic terminals were observed on cellular and axonal targets in the CNc. Small cells containing 70 nm dense-core vesicles received serotonergic input on their perikarya. Labeled synapses were also found on unlabeled axon terminals which made axoaxonal synapses on neuroendocrine processes. Non-synaptic terminals may be responsible for a variety of serotonin-mediated effects in the CNc. Synaptic interactions with local catecholaminergic and afferent cholinergic inputs to the CNc are likely.

Notes: Summary In an attempt to establish maturational alterations in the morphology of the articular tissue layer, mandibular condyles of four immature and four mature male monkeys (Macaca fascicularis) were studied using light microscopy as well as scanning and transmission electron microscopy. Specimens were fixed in situ by perfusion in the presence of ruthenium red to stabilize proteoglycans. Preparations intended for observation in the scanning electron microscope were first dehydrated and sputtered for the examination of articular surfaces, and afterwards treated with trypsin to expose the spatial arrangement of collagen fibrils. Gross anatomical relations between joint components indicated that the anterior and central, but not the posterior region of the condylar articular surface can be subject to compressional load. Load-bearing and non-load-bearing regions differed with respect to the morphology of the articular layer. Load-bearing surfaces were covered by a prominent articular surface lamina similar to that observed on articular cartilage. This lamina seemed to constitute an integral part of the articular layer, distinct from the lining of synovial fluid, and to be composed largely of proteoglycans. It was unaffected by maturation. The subjecent, load-bearing articular layer differed markedly in structure, both from articular cartilage, and between immature and mature animals. Articular cells of immature animals were classified as fibroblastlike, but unlike typical fibroblasts, were surrounded by a thin, often incomplete halo of fibril-free pericellular matrix, presumably consisting of proteoglycans. In mature animals, articular cells closely resembled chondrocytes, but exhibited prominent nuclear fibrous laminae, which usually are found only in fibroblasts. Thus, the load-bearing part of the articular layer seems to undergo a maturation-dependent metaplastic conversion, from a dense connective tissue with some features of fibrocartilage, to a fibrocartilage-like tissue containing chrondrocytelike cells with some features of fibroblasts. This conversion might reflect an adaptation to a maturation-associated increase in articular stress.

Notes: Summary Eleven dysplastic melanocytic naevi with various degrees of dysplasia, as judged by light microscopy, were studied by transmission electron microscopy, and their intra-epidermal melanocytes compared with those of five superficial spreading melanomas and seven common benign naevocellular naevi. Intra-epidermal melanocytes in dysplastic naevi exhibited signs of cellular atypia, which were most pronounced in the dysplastic naevi with histological high-grade dysplasia. A correlation between the degree of dysplasia at the light microscopic level and the degree of cytological atypia at the ultrastructural level was noted, and melanocytes in dysplastic naevi with a high degree of dysplasia had ultrastructural features similar to the melanocytes in superficial spreading melanomas. Our observations support the concept that dysplastic naevi fill the biological gap between benign naevocellular naevi and malignant melanomas and suggest that at least some of the dysplastic naevi must be regarded as potential precursor lesions of malignant melanoma, particularly those exhibiting a high degree of histological dysplasia.

Notes: Summary The skin of 20 patients with variegate porphyria (VP) was studied using light, fluorescent, and electron microscopy. Twelve patients had skin symptoms and markedly increased fecal protoporphyrin excretion. Their sun-exposed skin was characterized by homogeneous PAS-positive thickening and IgG deposition in the vessel walls. The basic ultrastructural change was thickening of the vascular walls caused by reduplication of the basal lamina and perivascular deposition of amorphous material. Qualitatively similar but less prominent histopathological changes occurred in sun-protected skin in some of the patients. Six patients had no skin symptoms but an increased porphyrin excretion. The light microscopical changes were comparable to those in the patients with skin symptoms, but the ultrastructural changes were less severe. No abnormal histopathological changes occurred in two symptomless patients who had low lymphocyte protoporphyrinogen oxidase activity but normal fecal porphyrin excretion. These results show that the primary site of skin damage in VP is the vessel wall, and that histopathological changes of the skin also occur in porphyric patients who have never had skin symptoms. Factors determining the occurrence of skin symptoms in VP are discussed.

Notes: Summary Ultrastructural changes of the connective tissue sheath (CTS), including the hyaline membrane, of human hair follicles during the hair cycle, were studied in normal scalp skin specimens. In early anagen, the CTS was composed of a thin basal lamina and surrounding collagen tissue. The collagen tissue gradually thickened during the development of the hair and hair follicle. In mature anagen hair follicles, the collagen tissue was separated into three layers. The inner collagen layer, just outside the basal lamina, was thin and composed of collagen fibres running longitudinally parallel to the hair axis. The middle collagen layer was very thick with its collagen fibres running transversely against the hair axis and surrounding the inner hair tissue. Many fibroblasts were present among the collagen fibres in the middle layer, whereas the inner layer contained almost none. In the outer collagen layer, collagen fibres ran in various directions parallel to the outer surface of the outer root sheath cells. In late anagen, the basal lamina became very thick. In catagen, the basal lamine and the inner collagen layer became corrugated and showed oedematous change and degeneration. Surrounding fibroblasts showed active production of new collagen fibres, which seemed to fill the spaces left by the retraction of the hair follicle and hyaline membrane. These ultrastructural changes of the CTS show that there may be dynamic metabolic changes of the connective tissue around human hair follicles during the hair cycle.

Notes: Abstract A many-called magnetotactic prokaryote obtained from brackish water was observed to possess intercellular connections at points of contact between the outer membranes of constituent cells. Each aggregate organism consisted of 10 to 30 individual Gram-negative cells containing material with the appearance of poly-β-hydroxybutyrate and magnetosomes of unusual arrangement, structure and composition. The aggregate, which possessed prokaryotic-type flagella arranged at the outwards surfaces of each cell, showed motility indicative of co-ordination between individual component cells. These results suggest that this organism could be a multicellular prokaryote.

Notes: Summary This study was designed to examine the vascular system of human xenotransplanted tumors on nude mice with different complementary morphometrical and morphological methods. The vascular system shows a chaotic arrangement. There is an extreme heterogeneity in the vascular distribution and density. Large avascular regions could be identified in several non-necrotic tumors. There was no clear difference in the vascular density between the center and the periphery of the tumors, nor was there any zonal correlation for the distribution of the necrosis. With three-dimensional corrosion casts it could be demonstrated that clusters of vessels were directly beneath areas almost free of vessels. In the center, vessels often form a sinusoidal system with numerous blind ends without clearly discernible endothelial cells. Numerous irregular tumor-cell-lined sinusoids are visible next to endothelial-lined vessels with transmission electron microscopy. With scanning electron microscopy it could be demonstrated that large-calibre endotheliazed vessels were found in the direct vicinity or in the center of non-viable zones. Even large-calibre vessels have a capillary wall structure. Sometimes, a basement membrane cannot be observed at all or only incompletely. There are numerous indications of vascular discontinuities and leaks with a widespread intercellular occurrence of blood cells.

Notes: Summary The present study was undertaken to define the effects of lipoprotein-derived cholesterol and endogenous, de novo synthesized cholesterol on the ultrastructure and function of undifferentiated rat adrenocortical cells [lipoprotein (HDL3 and LDL) receptor-negative, zona glomerulosa-like adrenocortical cells] in primary culture. For this purpose human plasma high density lipoprotein (HDL3) or low density lipoprotein (LDL) was added to culture medium devoid of cholesterol. Steroid secretion remained at the low basal level even after addition of lipoproteins, and the amount of intracellular lipid droplets did not increase. When mevinolin (0.96 µg/ml), an inhibitor of cholesterol synthesis, was added to the culture medium, a low secretion of corticosterone was measured both in serum-free and serum-containing media. Ultrastructurally, lipid droplets disappeared after treatment with mevinolin in both media used. At this concentration of mevinolin cell proliferation was similar to that in the controls, but at higher concentrations (4.8 or 9.6 µg/ml) proliferation was inhibited to 42% and 26% in serum-free medium, and 20% and 12% in serum-supplemented medium, respectively. This study demonstrates that cell proliferation and synthesis of corticosterone by undifferentiated rat adrenocortical cells is identical in the absence or presence of exogenous lipoprotein cholesterol. Inhibition of de novo cholesterol synthesis by mevinolin over a period of 7 days does not inhibit corticosterone secretion or proliferation of cells but decreases the amount of intracellular lipid droplets, thus suggesting utilization of intracellular cholesterol esters. However, higher concentrations of mevinolin inhibit proliferation of cells both in serum-free and serum-containing media.

Notes: Summary The phyllobranchiate gills of the green shore crab Carcinus maenas have been examined histologically and ultrastructurally. Each gill lamella is bounded by a chitinous cuticle. The apical surface of the branchial epithelium contacts this cuticle, and a basal lamina segregates the epithelium from an intralamellar hemocoel. In animals acclimated to normal sea water, five epithelial cell types can be identified in the lamellae of the posterior gills: chief cells, striated cells, pillar cells, nephrocytes, and glycocytes. Chief cells are the predominant cells in the branchial epithelium. They are squamous or low cuboidal and likely play a role in respiration. Striated cells, which are probably involved in ionoregulation, are also squamous or low cuboidal. Basal folds of the striated cells contain mitochondria and interdigitate with the bodies and processes of adjacent cells. Pillar cells span the hemocoel to link the proximal and distal sides of a lamella. Nephrocytes are large, spherical cells with voluminous vacuoles. They are rimmed by foot processes or pedicels and frequently associate with the pillar cells. Glycocytes are pleomorphic cells packed with glycogen granules and multigranular rosettes. The glycocytes often mingle with the nephrocytes. Inclusion of the nephrocytes and glycocytes as members of the branchial epithelium is justified by their participation in intercellular junctions and their position internal to the epithelial basal lamina.

Notes: Summary An ultrastructural study was made of the neurons, satellite cells and vesiculated axons of the intestinal nerve of the domestic fowl. Broad membrane-to-membrane contacts between adjacent nerve cell bodies were sometimes observed. The cell bodies and processes were not always separated from the extracellular space by a capsule of satellite cells. Following fixation using potassium permanganate, catecholamine (CA)-containing neurons in the intestinal nerve, unlike those in the lumbar parasympathetic ganglia, did not possess any small granular vesicles (SGV). Following exposure to noradrenaline, SGV could be demonstrated in the cell bodies of the juxta-ileal ganglia but not the juxta-rectal ganglia of the intestinal nerve. Non-CA axons were examined in tissue from birds that had been pretreated with 6-hydroxydopamine. Approximately one half of the non-CA axons formed axo-somatic contacts. Most of the non-CA axons contained varying proportions of small clear vesicles, large clear vesicles and large granular vescles. Statistical analysis showed that the non-CA axons could not be subdivided according to their vesicle content. CA-axons contained many SGV and were found in close apposition to neuronal somata and processes, and in the neuropil.

Notes: Summary An ultrastructural study of enterochromaffin-like (ECL) cells in the gastric mucosa of the white-belly opossum Didelphis albiventris (Marsupialia) was carried out. In parallel, histochemical methods were used at the light-microscopical level to demonstrate argentaffin cells, argyrophilic cells, and serotonin- and histamine-immunoreactive elements. Argentaffin and serotonin-immunoreactive cells were scattered, and argyrophilic cells were numerous, within the full thickness of the mucosa. Argyrophilic cell distribution was similar to that of histamine-immunoreactive elements. At the electron-microscopical level, the oxyntic mucosa of D. albiventris presented endocrine cells with secretory granules morphologically similar to those of the ECL cell of eutherian mammals. However, in this marsupial, the ECL cell exhibited a variable mixture of two distinct types of secretory granules: (1) granules with the morphological appearance of the eutherian ECL cell, and (2) granules morphologically similar to those of the eutherian enterochromaffin (EC) cells. Based on this morphological pattern of the ECL cell granules, it is proposed that in the oxyntic mucosa of the opossum D. albiventris, the EC and ECL cells represent distinct steps in the same line of cell differentiation; the ECL cell should also be a site of histamine storage.

Notes: Summary Mevinolin, an inhibitor of cholesterol synthesis, was used to study the effect of endogenous cholesterol synthesis on the morphology and function of differentiating and differentiated fetal rat adrenocortical cells grown in primary culture. Upon adrenocorticotrophic hormone (ACTH) stimulation under conditions in which endogenous cholesterol synthesis was inhibited but exogenous (lipoprotein) cholesterol was available, the cells differentiated normally from glomerulosa-like to fasciculata-like cells; the steroid hormone secretion was maximally induced. Under conditions in which cholesterol synthesis was maximally inhibited by mevinolin and the cells had no access to exogenous cholesterol, the cells did not differentiate into fasciculata-like cells; the ACTH-induced steroid response was highly suppressed under these conditions. The addition of either human low-density lipoprotein (LDL) or high-density lipoprotein (HDL3) to the culture medium restored the ACTH-induced differentiation and steroid secretion. Thus, in the absence of exogenous cholesterol, endogenous cholesterol synthesis was a prerequisite for differentiation. In cultures grown in the presence of exogenous cholesterol and ACTH with mevinolin-inhibited cholesterol synthesis and high steroid output, an increase in cytoplasmic lipids was evident, suggesting upregulation of LDL and HDL receptors. The results also demonstrated that induction of phenotypic differentiation from glomerulosalike into fasciculata-like cells can proceed in the presence of a cholesterol synthesis inhibitor like mevinolin; this differentiation in the absence of endogenous cholesterol synthesis is accompanied by the appearance of cytoplasmic cholesterol ester droplets, typical of fasciculata cells.

Notes: Summary Atrial appendage cardiocytes of mammals, including man, contain multiple cytoplasmic granules that vary in number in different physiological states. Using morphologic and comprehensive morphometric techniques, these granules were assessed in Sprague-Dawley rats following dehydration for 5 days, volume-loading by substituting 1% NaCl as drinking water for 7 days, unilateral nephrectomy plus volume-loading for 7 days, and in late term pregnant animals (18–20 days; term ≈21 days). Although principally located in the paranuclear region, granules were observed throughout the sarcoplasm. Cytological features indicative of synthetic activity and granule formation were readily apparent in all groups with the exception of pregnant rats where they were infrequently observed. Granule contents were released by exocytosis and observed in the right appendage of control, dehydrated and nephrectomy/volume-loaded groups and left appendage of volumeloaded animals. Exocytosis was not observed in pregnant animals. By point counting, the proportional volume of cardiocytes occupied by granules (V v ) in controls was significantly greater for right than for left appendage (2.12±0.22% vs 1.29±0.16%; mean±SEM;p〈0.05). A significantly similar difference was found for nephrectomy/volume-loaded animals. There was no significant difference inV v for right appendage between the control and experimental groups; for left appendage there was a significant increase inV v to 2.42±0.09% (p〈0.05) for volume-loaded animals only. Estimation of the maximum diameter of granule profiles in control animals was 238±9 nm and 230±6 nm for right and left appendages, respectively. The profile diameters in the left appendages of dehydrated (202±9 nm) and pregnant (200±7 nm) animals were significantly (p〈0.05) less than those of the control animals. The morphometric findings did not correlate with predictions based upon published biochemical data. In the course of this study, a previously unreported bimembranous, circular to ovoid structure was observed in the cardiocyte sarcoplasm of all animals; the nature and function of this structure is unknown.

Notes: Summary Short photoreceptor cells of the lamprey retina exhibited a 30% increase in the width of the myoid process and a 20% increase in that of the axonal process during a 12-h light period, compared to the measurements obtained during a 12-h dark period. An increase in the amount of cytoplasm, dilation of ER cisterns, and swelling of the nucleus appeared to cause the enlargement of the myoid parts. Accumulation of synaptic vesicles occurred concurrently with a thickening of the axonal process. These morphological changes presumably represent a phase of the diurnal cycle and current synaptic activity of the short cell. By contrast, the long photorecpetor cell showed neither measurable changes nor any indication of “retinomotor movement”.

Notes: Summary The paraventricular organ (PVO) and the posterior recess organ (PRO) of two elasmobranch species, the spiny dogfish,Squalus acanthias, and the skate,Raja radiata, were investigated by use of scanning and transmission electron microscopy and immunocytochemistry employing a series of primary antisera. The PVO and PRO contained four types of cerebrospinal fluid (CSF)-contacting neurons. One type was free of secretory granules and projected a dendrite-like process into the ventricle. The other three types were distinguished according to the size of their secretory granules. The ventricular extensions of these cells were filled with secretory granules. By means of immunocytochemistry three types of CSF-contacting neurons were observed in the PVO and PRO. Type I contained only serotonin; type 2 displayed only somatostatin; type 3 was endowed with both serotonin and somatostatin. Type I dominated in the PRO, whereas type 3 was the most frequent in the PVO. The latter cells appear to be the site of origin of a loose tract formed by serotonin- and somatostatinimmunoreactive fibers projecting from the PVO into the neuropil of the PRO. Compact bundles formed exclusively by serotonin fibers were also shown to extend between the PVO and PRO. The basal processes of the CSF-contacting neurons of the PRO penetrated into the underlying neuropil. This neuropil is rich in synapses and can be regarded as an integrative area to which the basal processes of the local CSF-contacting neurons, serotonin and somatostatin fibers from the PVO, and fibers containing immunoreactive thyrotropin-releasing hormone of unknown origin, support a conspicuous input. The present findings indicate that the PVO and PRO of elasmobranchs are functionally integrated structures.

Notes: Summary Dopaminergic and peptidergic nerve fibers were simultaneously demonstrated with a double-labeling technique at the ultrastructural level. The first antibody, raised against tyrosine hydroxylase, was applied during the preembedding phase and visualized with the peroxidase method. The second antibody, raised against one of the peptides met-enkephalin, somatostatin or gonadotropin-releasing hormone (GnRH), was applied to the ultrathin sections and visualized with gold-labeled goat anti-rabbit IgG. The fibers of both categories were present in the zona externa of the median eminence, frequently contacting the basal lamina of the portal vessels. In addition, topographical relationships between different types of nerve fibers were observed in the perivascular areas, although there were no morphological signs of synaptic specializations. Using serial sections, it could be established that one GnRH-fiber contacted both a dopaminergic fiber and a fiber immunoreactive for met-enkephalin. The observations support earlier physiological data concerning the regulation of the hypothalamo-hypophyseal axis, with special emphasis on the release of neurohormones in the median eminence of the newt.

Notes: Summary Crystalloids were found in Sertoli cells of the testis of the three-toed sloth by examination at the lightand electron-microscopic levels. Needle-, or spindle-shaped crystalloids, varying in length, were located in the basal part of the Sertoli cells. They consisted of bundles of filaments each measuring ~ 11 nm in diameter. Several filaments were packed hexagonally to form a bundle. The center-to-center distance between individual filaments of a bundle was ~ 17 nm. Periodical lateral projections emanated from the filaments. Cross sections of crystalloids showed that the projections radiated from each filament in three directions, forming an equilateral triangle with a side length of ~ 15 nm. Scattered polyribosomes were found between and around the bundles.

Notes: Summary The organization of testicular interstitial tissue of the spinifex hopping mouse, Notomys alexis differs from that of other rodents. It comprises between 10.3% and 17.3% (average 15.0%) of the total testicular volume, and is variable in its organization both at different locations within the testis of the one animal and among different individuals. Abundant, closely packed Leydig cells are usually present; however, in some regions large, thick-walled blood vessels and extensive peritubular lymphatic spaces, often lacking an endothelium adjacent to the Leydig cells, are also prominent. The Leydig cells in contact with the large blood vessels and lymphatics, unlike those in regions where lymph is sparse, are not densely packed and sometimes contain numerous lipid droplets. Ultrastructure of Leydig cells is typical of steroid-producing cells; however, mitochondria are often extremely large, unusual in shape or bizarely arranged in relation to one another. Also electrondense bodies displaying a paracrystalline-like internal structure of parallel, electron-dense filaments arranged in a lattice pattern occur in the cytoplasm of many cells. The significance of these unusual ultrastructural features and the organization of the interstitial tissue remain to be determined conclusively, but may relate to steroid synthesis, secretion and uptake.

Notes: Summary The immunoreactivity of atrial natriuretic peptide and ultrastructure of cardiocytes were examined in 5 species each of freshwater and seawater teleosts, as well as in 2 species each of elasmobranchs and cyclostomes. Immunoreactivity was strong in the atria of Cyprinus carpio, Anguilla japonica and Conger myriaster, rather weak in atria of Channa maculata, Lepomis macrochirus, Salmo gairdneri, Oplegnathus fasciatus and Eptatretus burgeri, very weak in atria of Pagrus major, Trachurus japonicus and Triakis scyllia, and not detectable in atria of Hexagrammos otakii, Narke japonica and Lampetra japonica. The immunoreactivity of the atrial cardiocytes was generally stronger in freshwater than seawater fish. Ventricular immunoreactivity was detected only in 7 species, always being weaker than that observed in the atrium. Ultrastructurally, however, secretory granules were found in atria and ventricles of all species examined, being more frequent in the former than the latter. By radioimmunoassay, immunoreactive ANP was detected in the extracts of blood plasma and both atrial and ventricular tissues of all species examined. There were no statistically significant differences in the values between freshwater and seawater species.

Notes: Summary It has been shown that pulse perfusion of rat liver with a digitonin-containing medium results in a highly zonated hepatocyte permeabilization, allowing selective sampling of cytosolic constituents from periportal and perivenous (centrolobular) hepatocytes “in situ”. In the present paper we provide an ultrastructural evaluation of the perfusion method. Identical changes in hepatocytes from affected periportal and perivenous zones are found. Affected hepatocytes appear light (electron-lucent) in electron micrographs with a sharp transition to normal hepatocytes. The most conspicuous ultrastructural findings are: (1) transformation of the sinusoidal part of the light hepatocytes, the lipocyte processes and the endothelium of affected zones apparently unifying into a continuous layer dominated by disrupted plasma membranes and 7-nm filaments; (2) deposition of osmiophilic digitonin-cholesterol complexes along the sinusoidal plasma membranes of affected zones; and (3) reduction of the cytoplasmic matrix (cytosol) in the light hepatocytes, a dilation of the mitochondrial intermembrane space with a preserved mitochondrial matrix, and a dilation of cisternae of the granular endoplasmic reticulum. The ultrastructural findings are consistent with marker-enzyme activity measured in eluates from digitonin-perfused livers, except that lysosomes appear intact, apparently contrasting with the observed eluation of amyloglucosidase (Quistorff et al. 1985).

Notes: Summary To determine the effect of cholinergic secretagogue on the Harderian gland of rats, several light- and electron-microscopic parameters were morphometrically assessed at different time intervals after carbamylcholine injection. In controls, two types of glandular cells (type A cells having 40–55 large vacuoles per cell profile and type B cells containing 30–38 smaller vacuoles per cell profile) and myoepithelial cells were recognized. At 5 min after injection of carbamylcholine, when rats secreted “bloody tears”, many alveoli showing narrower lumina and exocytotic figures in both types of cells were observed. Some vacuoles, which were covered by thin cytoplasmic sheets, protruded into the alveolar lumina. However, there was no evidence of apocrine or holocrine secretion. At 30 min and 120 min after injection, most of the alveolar lumina were dilated, and a pronounced decrease in the number of vacuoles in the glandular cells was observed. At 300 min after injection, the secretory vacuoles in both cell types reaccumulated. Transitional forms between the two cell types were not observed. The two types of Harderian gland cells can therefore be considered independent populations rather than different secretory stages of the same cell. It appears that the secretory process of the Harderian gland of rat is affected by cholinergic stimulation of the two types of glandular cells and of myoepithelial cells.

Notes: Summary Quantitative ultrastructural studies on the effects of visible and ultraviolet radiation on zooxanthellae in culture and in situ showed an inverse relationship between the volume fraction of chloroplast and irridiance. An independent effect of ultraviolet radiation was detected in cultured zooxanthellae only. The volume fraction of chloroplasts in cultured zooxanthellae and zooxanthellae in situ were the same, while the surface density of thylakoid lamellae relative to chloroplast volume in cultured zooxanthellae was less than in zooxanthellae in situ for all irradiances. Additionally, zooxanthellae in situ showed an effect of ultraviolet radiation on surface density of thylakoid lamellae. The response to different irradiances suggests a limit to photoadaptation by means of changing chloroplast volume, and that changes in thylakoid density are responsible for the continued photoadaptive plasticity observed. Flow cytometry and stereological studies show that the volume fraction of accumulation bodies within zooxanthellae increases with irradiance and ultraviolet radiation. Ultrastructurally, accumulation bodies do not resemble plant peroxisomes or glyoxysomes, while other inclusions observed in this stydy are suggestive of peroxisomes. This evidence suggests that accumulation bodies are not peroxisomes, but does support the previous assumptions concerning their role in autophagic processes.

Notes: Summary The fine structure of the muscle of the urinary bladder in female rats is similar to that of other visceral muscles, although it is arranged in bundles of variable length, cross-section and orientation, forming a meshwork. When distended, the musculature is 100–120 μm thick, with some variation and occasional discontinuity. Extended areas of cell-to-cell apposition with uniform intercellular space occur between muscle cells, whereas attachment plaques for mechanical coupling are less common than in other visceral muscles. There are no gap junctions between muscle cells. Many bundles of microfilaments and small elastic fibres run between the muscle cells. After chronic partial obstruction of the urethra, the bladder enlarges and is about 15 times heavier, but has the same shape as in controls; the growth is mainly accounted for by muscle hypertrophy. The outer surface of the hypertrophic bladder is increased 6-fold over the controls; the muscle is increased 3-fold in thickness, and is more compact. Mitoses are not found, but there is a massive increase in muscle cell size. There is a modest decrease in percentage volume of mitochondria, an increase in sarcoplasmic reticulum, and no appreciable change in the pattern of myofilaments. Gap junctions between hypertrophic muscle cells are virtually absent. Intramuscular nerve fibres and vesicle-containing varicosities appear as common in the hypertrophic muscle as in controls. There is no infiltration of the muscle by connective tissue and no significant occurrence of muscle cell death.

Notes: Summary Chronological changes in renal glomerular morphology and plasma renin activity were investigated during active and hibernating periods in the golden-mantled ground squirrel Spermophilus lateralis. The objective of this study was to determine whether the glomerular endothelium, visceral epithelium (podocytes), basement membrane, mesangial cells, proximal convoluted tubule cells and plasma renin activity exhibit measurable sequential differences between as well as within active and hibernating states at various time points. Limitations in the size of the experimental population prevented an evaluation of changes in these parameters during other important periods such as periodic arousal between hibernation bouts. In this study, glomerular endothelial pore number and epithelial filtration slit number significantly decreased by early hibernation when compared to those during summer activity, and then they increased back toward summer levels by late hibernation. In contrast, podocytic pedicel width along the glomerular basement membrane increased from summer activity to early hibernation, before significantly decreasing again by late hibernation. Mesangial cell and proximal convoluted tubule cell activity appeared increased during hibernation as compared to summer activity, whereas the width of the glomerular basement membrane showed no significant alterations throughout. Plasma renin activity significantly increased during early hibernation and mid-hibernation when compared to summer levels but had decreased by late hibernation toward summer values. The glomerular and plasma renin activity changes observed in this study clearly illustrate the drastic structural and functional adjustments which hibernating species make during torpor and also correlate well with the reported decrease in renal perfusion pressure and urine formation during hibernation. The observed morphological changes during hibernation do not appear to be temperature-dependent, because significant alterations in most of the parameters studied occurred during this period despite the fact that cold-room temperatures were kept constant throughout. The chronological approach to this study and its morphometric evaluation represent a pilot attempt at accurately documenting these changes during two critical states in the hibernator's cycle and may eventually lead to the characterization of these changes during the entire circannual cycle.

Notes: Summary The freshwater dinoflagellateGymnodinium acidotum is known to harbor a cryptomonad endosymbiont whose chloroplasts give the organism its blue-green coloration. Every cell examined from a wild population possessed chloroplasts, mitochondria, and other organelles which are of endosymbiotic origin. Transmission electron microscopy and fluorescence microscopy revealed that only 33% of these cells possessed the nucleus of the endosymbiont. The lack of a cryptomonad nucleus in some cells did not appear to affect the cells' ability to photosynthesize or move in response to varying levels of illumination. This represents the first report of a host/endosymbiont relationship in which a significant number of individuals from a given population lack a major endosymbiont organelle.

Notes: Summary The feeding behaviour of the rumen ciliatePolyplastron multivesiculatum has been studied with scanning and transmission electron microscopy. In contrast to other large entodiniomorphs,Polyplastron scarcely attaches to plant substrata such as straw or alfalfa; however, it readily phagocytes cellulose fibres (like epidermal bristles of alfalfa) suspended in the rumen fluid. The different stages of ingestion and intracellular degradation of this particular substrate are described at the ultrastructural level. Engulfment involves active movements of the vestibular lips. The digestion follows a peculiar pattern (“permeative” way): the cellulose substrate is gradually decomposed in situ inside the primary digestive vacuole without large fragmentation or pinching off of small vesicles. Thus degradation products are not stored in secondary vesicular systems as they are for other plant substrates (“cytotic” way). Thus, there is no single or general pattern of cellulosic structures lysis in entodiniomorph rumen ciliates. Rather, several pathways can be observed, probably according to the nature of the ingested plant material.

Notes: Summary The cellular interaction ofTetragoniomyces uliginosus andRhizoctonia sp. was restudied by transmission electron microscopy. During the first stages of interaction a body of medium electron density is visible at the center of the haustorial apex in close association with the plasmalemma. A single micropore is produced between the haustorial filament and the host cell. Cytoplasmic connection via the pore always occurred. The pore membrane is continuous with the plasmalemma of both cells. The protoplasts of both the haustorium and the host cell fuse via the micropore. An electron transparent to dense body occlude the pore. Among basidiomycetes, direct connection between the parasite and host protoplasts represents a hitherto unknown type of parasitic interaction.

Notes: Summary The effect of selenium deprivation onDaphnia magna was examined under controlled rearing conditions in a synthetic culture medium. After three generations, fertility was significantly reduced in deprived (Se−) animals. Growth and mortality of parent daphnids and development of parthenogenetic eggs were not affected during this period. In the fourth generation Se− daphnids rejected parts of their second antennae. At the ultrastructural level antennal muscle tissue was severely affected. Animals deprived of selenium had mitochondria and sarcoplasmic reticulum with myelin-like alterations. Giant lysosomes were present and complete lysis of muscle fibrils was observed in antennal muscle cells. These alterations are characteristic features of peroxidic damage in tissues. This interpretation is consistent with the function of selenium as a constituent of the enzyme glutathione peroxidase which protects cells from peroxidation. Selenium should be included in synthetic culture media for daphnids.

Notes: Summary Freeze substitution proved to be a valuable technique for studying the early stages of ascosporogenesis inAscodesmis nigricans. Our observations indicate that the ascus vesicle originated from the ascus plasma membrane. Invaginations of the plasma membrane produced ascus vesicle initials consisting of two closely spaced unit membranes. The appearance of the outer leaflet of each of these membranes was identical to that of the inner leaflet of the ascus plasma membrane. Apparent points of continuity between ascus vesicle initials and the plasma membrane were observed. Ascus vesicle initials accumulated in the ascus cytoplasm near the plasma membrane and then coalesced to form the ascus vesicle, a peripheral, cylinder-like structure consisting of two closely spaced unit membranes that extended from the ascus apex to the ascus base. The ascus vesicle then became invaginated in a number of regions and subsequently gave rise to eight sheet-like segments, or ascosporedelimiting membranes, that encircled uninucleate segments of cytoplasm forming ascospore initials. Like the ascus vesicle, each ascospore-delimiting membrane consisted of two closely spaced unit membranes, the inner of which became the ascospore plasma membrane. The ascospore wall then developed between the spore plasma membrane and the outer membrane. Many details of ascospore maturation were clearly visible in freeze substituted samples.

Notes: Summary Exposure of growingMicrasterias cells to high (32°–36°C) and low (3°–10°C) temperatures produces changes in morphology that are accompanied by several ultrastructural alterations. Whereas low temperatures essentially cause simplification of cell ornamentation, a variety of cell malformations result from high temperature treatment. These are the loss of cell symmetry leading to markedly aberrant cell shapes and an increase of “main lobes” with reduced degree of differentiation. Preliminary studies indicate that a shift in the distribution of membrane-associated Ca2+ by elevated temperatures probably underlies these abnormal cytomorphogenetic events. Both, low and high temperature cause a reduction in size of the young half cell and affect cytoplasmic streaming. Moreover, nuclear migration is retarded and chloroplast arrangement is influenced by temperature treatment at both ranges. Growth velocity of primary wall responsible for cell shaping is increased at high and slowed down at low temperatures compared to cells grown at 20°C. The main ultrastructural alterations induced by high temperatures are an increase in amount and length of ER cisternae, the appearance of “heat shock granule” aggregations localized in the cytoplasm, a reduced number of ribosomes and polysomes, the presence of oil bodies in growing cells and a varying thickness of the primary wall. Influences of low temperatures on ultrastructure are less pronounced. They are manifested in the formation of large aggregations of ER cisternae slightly differing from those found in untreated cells, a disturbed arrangement of the microtubule system surrounding the nucleus and a decrease of the number of cell wall forming cytoplasmic vesicles. It is thought that most of the temperature effects are due to an influence on membranes probably an alteration of ionic currents and, in addition, a modulation of normal protein synthesis.

Notes: Summary The dorsal and subventral esophageal glands and their secretory granules in the root-knot nematodeMeloidogyne incognita changed during parasitism of plants. The subventral esophageal glands shrank and the dorsal gland enlarged with the onset of parasitism. While secretory granules formed by both types of glands were spherical, membrane-bound, and Golgi derived, the granules differed in morphology and size between the two types of glands. Subventral gland extensions in preparasitic second-stage juveniles were packed with secretory granules which varied in diameter from 700–1,100 nm and had a finely granular matrix. Within the matrix of each subventral gland granule was an electron-transparent core that contained minute spherical vesicles. The size and position of the core varied within different granules. Few granules were present in the dorsal gland extension in preparasitic juveniles. The matrix of dorsal gland secretory granules formed during parasitism was homogeneous and more electron-dense than the matrix of subventral gland granules. Subventral gland secretory granules of parasitic juveniles and adult females appeared degenerate.

Notes: Summary Secretory activities of bicellular microhairs from grasses belonging to the subfamilies Chloridoideae, Arundinoideae, Panicoideae, and Bambusoideae, and including the “chloridoid”, “panicoid” and “Enneapogon” microhair morphological types, have been investigated. Light microscopic histochemistry indicated that all microhairs studied secrete polysaccharide and protein (or glycoprotein), including those which also secrete salt. Localization of polysaccharide at ultrastructural level using periodic acid-thiocarbohydrazidesilver proteinate staining revealed that in “panicoid type” microhairs dictyosomes are involved in polysaccharide secretion, whereas in the “chloridoid” and “Enneapogon” types “partitioning membranes” seem to be involved instead.

Notes: Summary Flagellar scales from the green flagellateTetraselmis striata (Prasinophyceae) were isolated, purified by isopycnic cesium chloride-gradient and zonal sucrose gradient centrifugation and their structure and biochemical composition investigated. Three types of flagellar scales were purified to more than 90% purity, a fourth type up to 75% purity. In addition to the previously known types of flagellar scales (pentagonal scales, rod-shaped scales, hair-scales), a novel scale type (i.e., the knotted scales) was discovered. New information about the asymmetric structure of the rod-shaped scales is presented and consequently they are renamed “man scales”. Flagellar scales consist mainly of carbohydrate (50–70%), significant amounts of protein (11% of dry weight) were found only in pentagonal scales. The main sugars (90%) of the pentagonal and man scales are the unusual 2-keto-sugar acids 3-deoxy-5-O-methyl-2-octulosonic acid (5 OMeKDO), 3-deoxy-2-heptulosaric acid (DHA), and 3-deoxy-2-octulosonic acid (KDO), the knotted scales contain as major sugars galactose and arabinose in addition to KDO and 5 OMeKDO but lack DHA. 13 major polypeptides were identified in flagellar scales by one-dimensional SDS-PAGE, 11 of these are of high molecular mass (〉116 kDa). While the majority of polypeptides was found associated with pentagonal scales, at least 4 polypeptides were tentatively assigned to the hair-scales and knotted scales.

Notes: Summary The vestibular sensory epithelia from two patients with acoustic neurinomas were examined ultrastructurally. Both patients had first undergone posterior fossa operations 8 months and 3 years before residual tumors in the acoustic canals were removed by a secondary translabyrinthine operation. Labyrinthine sensory tissues were also removed for microscopic studies. In case 1, the utricular macula and the lateral and anterior cristae were observed, with all sensory epithelia showing fairly normal findings. Myelinated nerve fibers below the sensory epithelia also appeared normal. In case 2, only the utricular macula could be observed. The sensory epithelium showed severe degeneration, disappearing sensory cells and increasing cytoplasmic filaments of both the sensory cell and the supporting cell. Myelinated nerve fibers below the sensory epithelia were only rarely found.

Notes: Summary Cations were precipitated with potassiumpyroantimonate in the middle ear mucosa of the rat and the distribution of the formed precipitates was studied by electron microscopy. The precipitate density in various cells of the different epithelia of the middle ear mucosa was determined on electron micrographs by counting the number of precipitates per unit area. Electron spectroscopic imaging was also performed to obtain information about the spatial distribution of the precipitates and their elemental composition.

Notes: Summary Kartagener's syndrome has been characterized by a primary ultrastructural abnormality of the cilia which consequently impairs their movements. We used transmission electron microscopy with tannic acid staining to investigate the fine structure of the cilia from the nasal mucosa of a 7-year-old girl with Kartagener's syndrome. The staining technique employed was useful for visualizing the dynein arms and protofilaments of the microtubules of the cilia. Although 15% of the cilia examined demonstrated microtubular disarrangements, these findings were considered to be acquired changes due to chronic sinusitis. No abnormal ciliary ultrastructures specific to Kartagener's syndrome, such as absence of dynein arms, were detected in this study. In such cases without any abnormal ciliary ultrastructures, it is conceivable that some other unknown factor may be involved in the impaired ciliary movement.

Notes: Synopsis At high latitudes, such as in Iceland, the daily photoperiod varies from almost continuous darkness in winter to virtually constant light in summer. Previous studies of detailed retinal structure in vertebrates have shown significant daily and annual effects of photoperiod. We sampled arctic charr in Iceland during the summer, including fish that were both light- and dark-adapted, during both day and night. We observed retinomotor responses characteristic of light- and dark-adaptation, but found no difference in the number of synaptic ribbons in the retina. The morpho-physiological changes, appearing as retinomotor responses, are thus not expressed at the synaptic level.

Notes: During postnatal development of the pigeon, a large portion of the skeleton becomes pneumatized, displacing the hemopoietic bone marrow. The consequences of pneumatization on distribution and quantity of bone marrow as well as the availability of other sites for hemopoiesis have been investigated. Hemopoietic marrow of differently aged pigeons divided into five groups from 1 week posthatching (p.h.) up to 6 months p.h. was labeled with Fe-59 and examined by serial whole-body sections. Autoradiography and morphometry as well as scintillation counts of single bones and organs were also carried out. No sign of a reactivation of embryonic sites of erythropoiesis was found. Bone marrow weight and its proportion of whole-body weight increased during the first 4 weeks p.h. from 0.54% to 2.44% and decreased in the following months to about 1.0%. The developing bone marrow showed a progressive distribution during the first months of life, eventually being distributed proportionally over the entire skeleton, except for the skull. At the age of 6 months p.h. bone marrow had been displaced, its volume decreasing in correlation to increasing pneumaticity and conversion to fatty marrow. This generates the characteristic pattern of bone marrow distribution in adult pigeons, which shows hemopoietic bone marrow in ulna, radius, femur, tibiotarsus, scapula, furcula, and the caudal vertebrae.

Notes: Artificially incubated fertile eggs from wild alligators have a significantly better hatch rate than those of captive, pen-reared alligators, possibly due to differences in the morphology of the eggshells. We compared the morphology of eggshells of wild alligators to those of captive alligators living in semi-natural environmental pens. Lengths and widths of eggs were measured and volume was determined, assuming an ellipsoid shape. Eggs were also evaluated for the quality of the eggshell (the presence or absence of rough deposits). Pieces of shell were cut from unincubated eggs and from eggs incubated for 55 days (just before hatching) and examined by scanning electron microscopy. Open pores on the outer surface of the shells were counted and thickness of the pieces was measured from micrographs. Results indicated that the number of pores on eggshells was lowest in eggs of captive alligators with early embryonic death. The number of pores was intermediate in eggs with early embryonic death from wild alligators, and the number of pores was highest in eggs with full-term embryos from wild or captive alligators. It is suggested that decreased porosity of eggshells may be associated with early embryonic death, is more prevalent in captive animals, and may, therefore, be related to poor hatch rate among penreared alligators.

Notes: Adult hornets (Vespa orientalis; Hymenoptera, Vespinae) build the brood combs out of organic or mineral matter. The cement that serves to glue the building material together is secreted in their saliva, the latter hardening within seconds to form fibers or plates. This saliva-derived spittle overlies and unites the building particles laminally and vertically. The hornet larvae spin a cocoon within the brood cells, which is largely fastened to and supported by the cell walls and is composed of a network of silk fibers and interlinking flat surfaces. On the outside of the cocoon fibers are spherical button-like structures that are very rich in phosphorus. The chemical composition of the adult salivary cement and the larval cocoon fibers is similar: both contain the elements P, Mg, S, Cl, K, and Ca. The possible biological significance of these findings is discussed.Among social insects belonging to the Hymenoptera, there are three main groups that build a multitude of cells, namely, the paper wasps (Polistinae), the other social wasps or hornets (Vespinae), and the social bees (Apinae). The constructed cells serve mainly for rearing the brood, but in many instances (particularly among various Apinae) they also serve for storing honey and pollen. For building material, Apinae rely primarily on beeswax (a product secreted by the bee itself to which various amounts of plant resins are added). On the other hand, wasps (Polistinae and Vespinae) build mainly from matter collected in the nearby environment, be it organic matter such as tree bark, mineral matter, or a combination of the two.Much information has accumulated on comb and cell building among these insect groups (e.g., Wheeler, '23; Van der Vecht, '57, '65; Lindauer, '61; Michener, '61; Kemper and Döhring, '67; Wilson, '71; Guiglia, '72; Spradbery, '73; Edwards, '80; Brian, '83; Schremmer et al., '85). Species of Polistinae and Vespinae are prevalent in forest areas in Southeast Asia and in central and South America or in the temperate regions in both the northern and southern hemispheres, and they mainly use vegetable matter to build their combs. In contrast, species prevalent in the Mediterranean region, which is dry and relatively unvegetated during the wasp and hornet active season, rely more on nearby mineral than on plant matter. Comb building in the Oriental hornet is well known (Darchen, '64; Ishay et al., '67; Schaudinischky and Ishay, '68; Ishay, '73, '75a, b, '76; Ishay and Sadeh, '75, '76; Ishay and Perna, '79; Ishay et al., '82). Recently Ganor et al. ('86) described the cell wall in the V. orientalis comb, showing it to be composed of mineral particles collected near the subterranean nest, in contrast to the comb of two European species (V. crabro and Vespula) (Paravespula) germanica, which is built primarily of organic matter. Regardless of whether the building material is mineral or organic, the hornets utilize particles of sand or other minerals or pieces of wood wrapped and melded together by saliva. However, nothing is known about the cement or mortar used to glue together these “bricks.” The present investigation was initiated to increase our knowledge of this cementing substance.Combs of V. orientalis were collected from fields in the Tel-Aviv district in 1987 during the active season, which extends over most of the summer months. Because the combs are easily damaged, care was taken to remove them intact from the natural nest. Once removed, the combs were cleared of the existing brood (eggs, larvae, or pupae) and then stored carefully in dry glass vessels until examined. For the present study, combs were collected only from nests in Khamra soil, which is common in the Tel-Aviv district and along the coastal area of Israel. To investigate the morphology of the cement material in the comb, strips of comb wall as well as segments of the pupal silk dome were removed from each comb and prepared for examination. The comb wall strips were cut to a size of 3 × 6 mm and fastened to the stub of a scanning electron microscope (SEM). The silk pieces were processed in two ways: (1)strips of 3 × 4 mm were fastened onto a stub with the convex (outer surface) facing up or (2) with the concave (interior) side upward.Silk strips were boiled for 2 hours in distilled water to remove all impurities, such as foreign matter adhering to the silk secreted by the larvae. Examination of these silk fibres was carried out in the three ways: (1)SEM micrography of the exterior (white portion) of the silk dome; (2) micrography of the interior; (3) micrography of both exterior and interior aspects after boiling for 2 hours in distilled water to remove water-suspended foreign materials that may have clung to the silk dome in the course of ordinary nest activity.The chemical composition of selected specimens was investigated by x-ray analysis. They were done on JEOL 840 Scanning Electron Microscope (SEM) equipped with Link 10,000 Energy-Dispersive System (EDS). (With the EDS System, the spectrometer separates the elements according to energy rather than wavelength). Quantitative analysis was by ZAF4 program. Five strips of comb cell wall were examined from a randomly selected comb, each comb from a different nest. Micrographs were taken of isolated silk samples from each of the examined combs.

Notes: Dissections were performed to document buccal anatomy in three species of the pulmonate genus Helisoma Swainson, 1840. The 28 muscles which are responsible for radular feeding in these animals are organized in three concentric and integrated envelopes. The deepest of these includes muscles which manipulate the radula about the odontophoral cartilage. Elements of the middle envelope direct movements of the cartilage within the buccal cavity, and muscles of the outer envelope control movements of the buccal mass within the cephalic haemocoel. Motion analysis by videomicrography showed that muscles of the middle and outer envelopes contribute to the action of radular feeding by acting as antagonists to other muscles and to hydrostatic elements of the buccal apparatus. Observations of radular dentition showed that although each of the three species examined has a unique radula, especially with regard to the specific details of tooth shape, all resemble a radula characteristic of the Planorbidae with regard to other, more general, aspects of ribbon architecture.

Notes: Crocodilians and birds are the extant representatives of a monophyletic taxon known as archosaurs. Their limbs are highly derived in terms of reduction in number of skeletal elements in both the carpus and the tarsus. It is necessary to have a detailed description of crocodilian limb ontogeny to address the evolutionary issue dealing with the origin and organization of the avian limb. In this paper, we present an analysis of the early development of the crocodilian limb skeleton. Contrasting with earlier observations, we redefine the number and composition of carpal, tarsal, and phalangeal elements. This ontogenetic information is then used to introduce a revision of the homologies of the skeletal elements in the crocodilian limb. Some invariances are pointed out in the developmental organization of tetrapod limbs and this evidence serves to readdress several issues concerning the evolution of the avian limb. We present further embryological data in support of the hypothesis that digits 2-3-4 are the components of the wing skeleton in birds. In general, our comparative survey indicates that the elements that appear late in ontogeny are the ones lost in phylogeny. By comparing turtle (primitive) limb development with crocodilian and bird development, we propose a hypothesis in which the derived skeletal patterns found in crocodilians and birds have originated by a heterochronic process of paedomorphosis.

Notes: A unique pathway that utilizes endoplasmic reticulum (ER) networks is proposed for screening pigment granule formation in the retina of adult Eumesosoma roeweri. Smooth endoplasmic reticulum (SER) functions to transport pigment particles to the formative site. Each site is composed of concentric, interconnected rings of SER that are filled with dense-cored, spherical pigment particles. Formation of the screening pigment granule begins by the release of particles from the innermost rings of carrier SER. Continued release followed by fusion and condensation of the pigment particles results in the formation of a mature pigment granule.

Notes: Light and scanning electron microscopy of vascular replicas from the facultative air-breathing fish Heteropneustes fossilis show modifications in the macrocirculation of the respiratory organs and systemic circulation, whereas, gill microcirculation is similar to that found in typical water-breathing fish. Three and sometimes four ventral aortae arise directly from the bulbus. The most ventral vessel supplies the first pair of arches. Dorsal to this another aorta supplies the second gill arches, and a third, dorsal to, and larger than the other two, supplies the third and fourth arches and the air sacs. Occasionally a small vessel that may be the remnant of a primitive aortic arch arises from the first ventral aorta and proceeds directly to the mandibular region without perfusing gill tissue. The air sac is perfused by a large-diameter extension of the afferent branchial artery of the fourth gill arch and its circulation is in parallel with the gill arches. Blood drains from the air sac into the fourth arch epibranchial artery. A number of arteries also provide direct communication between the efferent air sac artery and the dorsal aorta. All four gill arches are well developed and contain respiratory (lamellar) and nonrespiratory (interlamellar and nutrient) networks common to gills of water-breathing fish. Air sac lamellae are reduced in size. The outer 30% of the air sac lamellar sinusoids are organized into thoroughfare channels; the remaining vasculature, normally embedded in the air sac parenchyma, is discontinuous. A gill-type interlamellar vasculature is lacking in the air sac circulation. Despite the elaborate development of the ventral aortae, there is little other anatomical evidence to suggest that gill and air sac outflow are separated and that dorsal aortic oxygen tensions are maintained when the gills are in a hypoxic environment. Physiological adjustments to hypoxic water conditions probably include temporal regulation of gill and air sac perfusion to be effective, if indeed they are so.

Notes: The antennae of adult Damalinia ovis, the sheep louse, were studied using light and scanning electron microscopy. Sensory structures are located on all three antennal segments with the predominant sensilla type being tactile. Nine different types of sensilla are described on the basis of external appearance. One of the sensilla, designated a “pit organ” because of its unusual shape, has not been described previously. A pair of these sensilla are present on each antenna, and their function is unknown. A group of 11 sensilla on the tip of each antenna contains olfactory and chemosensory pegs, and a possible thermohygroreceptor. The antennae are sexually dimorphic, the male having more tactile sensilla, two well-developed terminal hooks, and a different cuticular architecture on the posterior surface of antennal segment 1.

Notes: The formation of the unpaired structure ventral to the basibranchial region, the so-called urohyal, differs within osteichthyans. A cartilaginous preformed, unpaired “urohyal” is present in sarcopterygians. A three-tendon ossification is present in Polypterus. An “urohyal” or urohyal is absent in both Amia and Lepisosteus. The urohyal formed as an unpaired ossification of the tendon of the sternohyoideus muscle is a feature of teleosts. A new structure, the parurohyal, arises as a double ossification of the tendon of the sternohyoideus muscle in siluroids; during ontogeny an anterodorsal crest or cup-like structure derives from the anterior basibranchial region and the tendon bone; therefore, the parurohyal is compound in origin. Judging from their formation and their distribution within osteichthyans the cartilaginous preformed “urohyal” and the teleostean urohyal are nonhomologous, whereas the urohyal and parurohyal are homologous. The urohyal is connected by ligaments with the ventral hypohyals in most teleosts, whereas it articulates with the ventral hypohyals in teleosts such as Anguilla and Chanos. The parurohyal is a synapomorphy of siluroids. The parurohyal in siluroids is articulated with both ventral and dorsal hypohyals, and with the basibranchial region in catfishes such as diplomystids and ictalurids, whereas it articulates only with the ventral hypohyals in other catfishes such as trichomycterines. The passage of the hypobranchial artery through the hypobranchial foramen of the parurohyal is a unique feature of siluroids, like the absence of the basihyal bone.An ossified dorsal hypohyal appears late in ontogeny in Amia, as do tooth plates related to the medial side of the hyoid arch and branchiostegal rays in Amia, and tooth plates on the hyoid arch and branchiostegal rays in Elops (unique features within extant teleosts). Two ossified hypohyals, a synapomorphy of teleosts, are present early in ontogeny.There is intraspecific variation in the onset of ossification of the bones of the head, but the sequence of ossification between bones in a defined structural system is conserved (e.g., branchiostegal rays ossify first, then bones of the hyoid arch).

Notes: The vas deferens of Ambystoma macrodactylum is composed of a peritoneal epithelium, connective tissue layer with fibroblasts, circular smooth muscle, capillaries, cells containing lipid, and a luminal epithelium composed of a single layer of cuboidal cells covered by a net of interconnected ciliated squamous cells. The cuboidal cells have abundant rough endoplasmic reticulum, mitochondria, and PAS + secretory vesicles. Squamous cells of breeding males consistently have tufts of ∼100 cilia located at one end of the long axis of each cell. These cilia may help distribute secretory products. The squamous cells, absent in post-breeding males, are apparently sloughed into the lumen. Lipid vesicles are present throughout the cytoplasm of the cuboidal and squamous epithelial cells and are also in some cells of the connective tissue layer. These vesicles increase dramatically in number during the first 4 weeks after breeding and may serve as an energy pool for the next breeding season. Enzyme-histochemical tests for testosterone synthesis were negative. In addition to the accumulation of lipid and the loss of squamous cells in the vas deferens, after breeding PAS + vesicle production is terminated. These alterations appear to represent energy conservation strategies employed by the sperm-depleted vas deferens.

Notes: The concept of parallel muscle combinations, in which spindle density is significantly higher in small muscles compared to their larger counterparts in large-small muscle combinations acting across a joint, is supported by the results of this study regardless of the joint. Analysis of the canine data as well as previously published guinea pig forelimb and human pelvic limb data revealed no significant difference in spindle density between antigravity and non-antigravity muscles. Furthermore, a gradual increase in spindle density from proximal to distal on the limb was not found, although spindle density was significantly higher in the intrinsic manus or pes muscles compared to more proximal limb muscles in all three species. The significant differences in spindle densities in parallel muscle combinations and in manus/pes versus proximal muscles are discussed relative to their possible role in the control of locomotion.

Notes: The choanocyte chambers of eight different haplosclerid sponge species have been investigated with regard to their histological structure, their relation to the mesohyl, and their arrangement between the canal systems. Two fundamentally different organizational types occur in the investigated sponge species. In Haliclona rosea, H. oculata, H. elegans, H. simulans, H. rava, H. fistulosa, and Acervochalina limbata the choanocytes are separated from the mesohylar tissue, being more or less covered over the outer surface by pinacocytes belonging to the incurrent canal walls. In Haliclona indistincta, on the other hand, the choanocytes are, as in most other Demospongiae, in contact with the mesohyl at their outer surfaces. This indicates that the present order Haplosclerida is polyphyletic and contains sponges of a presumably poecilosclerid origin.

Notes: The three-dimensional structures of the carotid labyrinth in five species of anurans representing four families (Rana nigromaculata, Rana catesbeiana, Bufo japonicus, Hyla arborea, and Xenopus laevis), and three species of caudates representing three families (Cynops pyrrhogaster, Hynobius nebulosus, Ambystoma mexicanum) were compared using vascular corrosion castings and scanning electron microscopy (SEM). Anuran carotid labyrinths are spherical in shape and are classified into two groups according to the origin of the external and internal carotid arteries. One group, which included Rana, Hyla, and Bufo, is characterized by the presence of a vascular ring at the proximal end and some vascular routes at the distal end of the labyrinth. The external and internal carotid arteries originate from these structures. The other group, which includes only Xenopus, is characterized by the external carotid artery opening directly from the central chamber or the common carotid artery, and by the internal carotid artery originating from within the vascular maze. The vascular maze is most complex in Xenopus, less so in Rana and Bufo, and simplest in Hyla. The carotid labyrinths in Cynops and Hynobius are oblong in shape. The fundamental organization in salamanders is similar to that in anurans. The vascular maze, however, is much simpler than in Hyla. There is no specialized swelling in Ambystoma mexicanum. The present findings suggest that most amphibian carotid labyrinths have the appropriate architecture for controlling vascular tone.

Notes: The vomeronasal sensory epithelium of a crotaline snake, Trimeresurus flavoviridis, was shown to consist of a superficial supporting cell layer and an underlying sensory cell layer composed of columns of sensory cells. The supporting cell layer consists of both supporting cells and dendrites of the underlying sensory neurons. The apical regions of sensory cell dendrites contain numerous microtubules, many elongated mitochondria, centrioles, and electron-dense bodies. The dendrites terminate as dendritic knobs from which microvilli project into the vomeronasal lumen. Smooth vesicles are abundant in the dendritic terminals and their vicinity. Supporting cells also bear microvilli, and these cells contain large electron-opaque granules and dense vesicles near their free surfaces. Cytoplasmic extensions of the supporting cells form a meshwork which separates dendrxites from each other in the vicinity of the luminal surface. The meshwork becomes obliterated in the infranuclear region of each supporting cell. Bipolar-shaped sensory cells with lightly stained round nuclei contain the characteristic cell organelles of neurons and are thought to be sensory neurons. These cells are especially characterized by well-developed lamellae of rough endoplasmic reticulum and extensive arrays of smooth endoplasmic reticulum. The perikarya of cells located in the apical region of the cell columns tend to contain larger amounts of smooth endoplasmic reticulum and lipofuscin granules than the perikarya of cells located in lower regions. Undifferentiated cells are found in the basal region of the columns. Satellite cells form the framework of the columns and are also found among neuronal elements.

Notes: The protandric hermaphrodite Ophryotrocha puerilis possesses one pair of eyes. They are located in the peristomium. Each light-sensitive organ consists of one sensory cell and one to two supporting cell(s) embedded in a cup-shaped reflector. The sensory-supporting cell complex is enveloped by a basal lamina. This lamina is supposed to be identical with the neural lamella. Thus the eyes proper have to be regarded as protrusions from the brain, while epidermal cells seem to differentiate to crystalline cells (reflector) and are deposited onto the sensory complex.The reflector is built up by several cup-shaped cells (juveniles, 4-5; adults, 10-12). Each of these cells comprises a multilayer of parallel-oriented, membrane-bound crystalline platelets which are thought to be guanine. The sensory cell is of the inverted rhabdomere type. Submicrovillar cisternae, typical for most polychaete eyes, are lacking. The first and always present supporting cell entirely envelops the sensory cell, thus forming the extracellular space around the rhabdomere. It does not contain any pigment granules. Often but not always a second supporting cell has been observed surrounding the sensory cell and first supporting cell. It is interpreted as a glial cell.In the sensory cell beneath the rhabdomere, pino- and phagocytosis can be observed and secondary lysosomes are found in high densities. Preliminary results seem to demonstrate that there is no distinct diurnal cycle of receptoral membrane recycling. In comparison with the ocelli of Dinophilidae, which have been interpreted as a dorvilleid-related family, morphological differences and their application to phylogenetic considerations are discussed.

Notes: The appearance pattern of pharyngeal tooth germs was investigated in the larval Japanese dace, Tribolodon hakonensis, which has a bilaterally asymmetrical dentition. Teeth develop in a series of replacement waves beginning with the initial central tooth (Ce) and continuing with teeth of anterior (An) and posterior (Po) positions relative to the initial one. Identified by wave number (n) and tooth position (r), according to the formula n-1[r], tooth germs appeared in the order of tooth 0[Ce0], 1[Po1], 1[Anl], 2[Ce0], 2[An2], 3[Po1], 3[An1], 4[Ce0], 4[An2], 5[Po1], 5[An1], 5[An3], 6[Ce0], 6[An2] during the larval period. Dentition on the right side, however, lacks the first tooth at position An2 (tooth 2[An2]) and teeth at position An3. Tooth germs on the first, second, and third replacement waves appeared simultaneously on the arches of both sides. During following waves, tooth germs on the left side appeared later than those on the right. Delay of tooth germ appearance On the left side is interpreted as an inhibitory influence of existing tooth germs in accordance with Osborn's (Proc. R. Soc. Lond. Ser. B 179:261--289, '71) theory. The delay of tooth germ appearance on the left arch is most pronounced on the seventh replacement wave. Teeth of the right major row in adults of this species are replaced more frequently than those of the left major row, apparently in correlation with the absence of the first larval tooth at position An2 and teeth at position An3. It is hypothesized that cyprinids evolved the minor rows and specialized teeth of their adult dentition as apomorphic characteristics by the process of neoteny.

Notes: Development of the internal structures of the caudal region (muscles, bones, cartilages, nerves, and blood vessels) of the Da/Da mutant medaka and of wild-type +/+ fish was compared. Muscles and nerves were immunochemically stained by using an antibody to troponin T and antibodies to neurofilament proteins, respectively. Bones and cartilages were stained with alizarin red and alcian blue. In stages 31-32 of the Da/Da embryo, the rudiments of epurals 1, 2, and 3 differentiated dorsally opposite the sites of the ventral rudiments of the parhypural, the lower hypural plate, and the upper hypural plate, respectively. In the +/+ embryo, the rudiments of epurals 1 and 2 differentiated in the later developmental stages (after stage 34), but the rudiment of epural 3 did not differentiate. In the Da/Da embryo, the deep dorsal flexor passed dorsalwards and ended at the rudiment of epural 2, whereas the muscle passed ventralwards and ended at the rudiment of the upper hypural plate in the +/+ embryo. In the Da/Da fry, the epichordal region of the caudal fin fold grew and the notochord extended straight into the tail without bending, while in the + /+ fry, the epichordal region did not grow and the notochord turned dorsalwards within the tail. In the Da/Da fry, extra skeletal rudiments and musculature differentiated in the epichordal region of the caudal fin, whereas these supernumerary structures did not develop in the +/+ fry. The pattern of the epichordal skeleton and musculature in the caudal region of the adult Da/Da fish was an almost mirror-image duplication of the hypochordal structures. These observations suggest that the characteristic expansion of the dorsal fin and epichordal lobe in the caudal region of the adult Da/Da fish is due to a process of ventralization, whereby the dorsal skeleton, musculature, and fin fold are stimulated to differentiate in a pattern like that which is restricted to the anal fin and hypochordal lobe of the wild type.

Notes: Tongue musculature in 24 genera of snakes was examined histologically. In all snakes, the tongue is composed of a few main groups of muscles. The M. hyoglossus is a paired bundle in the center of the tongue. The posterior regions of the tongue possess musculature that surrounds these bundles and is responsible for protrusion. Anterior tongue regions contain hyoglossal bundles, dorsal longitudinal muscle bundles and vertical and transverse bundles, which are perpendicular to the long axis of the tongue. The interaction of the longitudinal with the vertical and horizontal muscles is responsible for bending during tongue flicking. Despite general similarities, distinct patterns of intrinsic tongue musculature characterize each infraorder of snakes. The Henophidia are primitive; the Scolecophidia and Caenophidia are each distinguished by derived characters. These derived characters support hypotheses that these latter taxa are each monophyletic. Cylindrophis (Anilioidea) is in some characters intermediate between Booidea and Colubroidea. The condition in the Booidea resembles the lizard condition; however, no synapomorphies of tongue musculature confirm a relationship with any specific lizard family. Although the pattern of colubroids appears to be the most biomechanically specialized, as yet no behavioral or performance feature has been identified to distinguish them from other snakes.

Notes: Wall and internal organelles of the encysting salt marsh heterotrich ciliate Fabrea salina were examined by bright field and Nomarski interference contrast microscopy and by transmission and scanning electron microscopy. A mucoid sheath believed to be derived from bacteria covers the ectocyst. The possible bacterial origin of this sheath has been demonstrated experimentally by its removal and reappearence after 24 hours. Control ectocyst maintained in sterile seawater did not replace the sheath. The ectocyst has a crinkled appearance. The endocyst is composed of two layers. The inner endocyst material is continuous with that of the plug located at one end of the cyst. The two membranes are separated from one another by an interwall space filled with fibrous material. The cytoplasm, covered by an outer double membrane pellicle, contains mucocysts, pigment granules, microtubules, non-ciliated kinetosomes, Golgi complexes, ribosome-studded endoplasmic reticulum, and mitochondria. The endoplasm contains numerous autophagosomes, mitochondria, and food reserve materials. The macronucleus is centrally located in the cytoplasm of the encysting organism.

Notes: Analysis of lateral and dorsoventral radiographic films shows that ingestion, transport, and mastication in Pedetes capensis (Rodentia) are cyclic and their movement patterns are essentially similar for the three food types offered. During the ingestion cycle, closing of the mouth is accompanied by a backward translation of the condyles, so that movement is predominantly orthal. During the opening stage, the extent of the anterior condylar translation is smaller. As a result the mandibular incisors move ventrally and posteriorly. During the ingestion cycles, food is transported to the back of the tongue, with the transverse rugae and the folds of the upper lip playing important roles.Springhares show a bilateral masticatory pattern; food is chewed on both sides simultaneously. During chewing, the condyles lie in their most forward position at maximum opening of the mouth. The mouth is closed by rotation of the lower jaw around the temporomandibular joint coupled with posterior condylar translation. At the beginning of the slow-closing stage, the upward rotation of the mandible slows and the jaw slowly shifts forward. During the grinding stage, the mandible is shifted forward with both toothrows in occlusion. During the opening stage, the jaw returns to its starting position.Comparison of kinematic and anatomical data on rodent mastication suggests that some dental characteristics form the most important factors regulating the masticatory pattern and consequently allow reasonably reliable prediction of rodent masticatory patterns.

Notes: Avian embryos can be completely paralyzed by injection of neuromuscular-blocking agents. We used a single injection of decamethonium iodide to paralyze embryos at 7, 8, or 10 days of incubation and analyzed the growth of individual bones (clavicle, mandible, ulna, femur, tibia, humerus) and of individual muscles that act upon some of those bones (clavicular and sternal heads of m. pectoralis, and mm. biceps brachii, depressor mandibulae, pseudotemporalis, and adductor externus). Growth of the bones is not equally affected by paralysis. Only 27% of clavicular growth (by mass) but 77% of mandibular growth occurred in paralyzed embryos, whereas the four long bones exhibited 52-63% of their normal growth. Analysis of muscle weight, fiber length and physiological cross-sectional area (weight/fiber length) indicate that there was greater reduction of the muscles acting on the limbs than of those acting on the mandible, i.e., diminished growth of the skeleton is correlated with reduced muscular activity. Specific retardation of clavicular growth is due to fusion of sternal rudiments and collapse of the thorax, as well as virtual absence of the musculature that normally attaches to the clavicle. We discuss these results in the light of intrinsic and extrinsic factors governing growth of tne embryonic skeleton. Paralysis reduces skeletal growth by reducing both the movements taking place in ovo, and the loads imposed on the bones by muscle contraction, changes that represent alterations in the mechanical environment of the skeleton.

Notes: This study using light and electron microscopy indicates that the saccus vasculosus is distinguishable in 9-mm embryos and grows continuously throughout embryonic development to the adult stage. In the saccus vasculosus, epithelial mitoses are observed in all stages studied. Phases of centriologenesis, ciliogenesis, and globule formation have been characterized in developing coronet cells. During the phase of centriologenesis, new centrioles appear in association with pre-existing centrioles and not on deuterosomes. After ciliogenesis, each cilium differentiates to a globule almost at the same time as the other cilia of the coronet cell. The inner membrane system of the globules seems to derive from the ciliary plasma membrane. This membrane system often produces membrane whorls during the development. The different phases of coronet cell development have been found in the same individual and in all the stages studied except the 9-mm embryo. Cerebrospinal fluid-contacting neurons are observed in the saccus epithelium from the 12-mm embryos on and are distinguishable from coronet cells in their early formative stages. The three cell types of the saccus vasculosus increase continuously in number during development. Nerve processes are found in the saccus vasculosus of embryos, whereas differentiated synapses appear later in the fry. The significance of continued coronet cell formation is discussed in relation to a putative coronet cell and/or a globule renewal cycle in the adult.

Notes: Changes of architecture of adult rat gastrocnemius medialis muscle (GM) due to growth were studied in relation to length-force characteristics. Myofilament lengths were unchanged, indicating constant sarcomere length-force characteristics. Number of sarcomeres within fibers was unchanged as a consequence of growth, allowing persistence of differences between proximal and distal fibers in all age groups. Distal fiber length at muscle optimum length was shorter for the 14- than for the 10- and 16-week age groups despite a lack of difference of number of sarcomeres. This is indicative of a shift of optimum fiber length with respect to muscle length to lengths higher than optimum length. Some evidence for the occurrence of distribution of fiber optimum lengths with respect to muscle optimum length was found in other age groups as well, albeit of a smaller magnitude. Muscle and aponeurosis length increased substantially with growth. Functional effects of increased aponeurosis lengths were increased contributions to muscle length changes by the aponeurosis, allowing smaller fiber contributions in older animals. Fiber angle increased approximately 5° with growth. Despite the differences of architecture indicated above, muscle length range between optimum length and active slack length was constant. This was probably caused by widening of this length range in the youngest age group by variations of architecture within the muscle. It is concluded that adaptation of aspects of muscle architecture is an important mechanism for adult muscle growth in rat GM. Of these aspects regulation of muscle length seems a dominant factor.

Notes: A light and electron microscopic study of the skin of domestic chickens, seagulls, and antarctic penguins revealed abundant extracellular dermal lipid and intracellular epidermal lipid. Dermal lipid appeared ultrastructurally as extracellular droplets varying from less than 1 μm to more than 25 μm in diameter. The droplets were often irregularly contoured, sometimes round, and of relatively low electron density. Processes of fibrocytes were often seen in contact with extracellular lipid droplets. Sometimes a portion of such a droplet was missing, and this missing part appeared to have been “digested away” by the cell process. In places where cells or cell processes are in contact with fat droplets, there are sometimes extracellular membranous whorls or fragments which have been associated with the presence of fatty acids. Occasionally (in the comb) free fat particles were seen in intimate contact with extravasated erythrocytes. Fat droplets were seen in the lumen of small dermal blood and lymph vessels. We suggest that the dermal extracellular lipid originates in the adipocyte layer and following hydrolysis the free fatty acids diffuse into the epidermis. Here they become the raw material for forming the abundant neutral lipid contained in many of the epidermal cells of both birds and dolphins. The heretofore unreported presence and apparently normal utilization of abundant extracellular lipid in birds, as well as the presence of relatively large droplets of neutral lipid in dermal vessels, pose questions which require a thorough reappraisal of present concepts of the ways in which fat is distributed and utilized in the body.

Notes: Development of neurons in the area triangularis of Gallotia galloti was investigated in Golgi-impregnated brain tissue. Four major neuronal types present in adults were found to originate from two migratory neuroblast types, which were followed from embryonic stage S.32. One type has a thick main medial process, whereas the second type has a long main lateral process. As they migrate toward the periphery of the nucleus, morphological characteristics of maturation appear, including growth cones, filopodia, and outgrowth of axons. Neuroblasts with a main lateral process differentiate into two immature neuronal types, bipolars and pyramidals, observed at S.33 and thereafter. The neuroblasts with a main medial process undergo some somatic translocation through a transitory tangential shaft. Then they develop into monopolar immature forms with a long varicose medial, process, appearing from S.36. onward. Immature bipolar neurons do not experience great changes in their dendritic arborization during development to the adult stage, but pyramidals and monopolars undergo a rapid development of the dendritic tree after S.36. By S.38 archetypes of adult neuronal forms are established. Hairlike appendages first appear on neurons at S.36 They decrease suddenly in S.38 and then proliferate in S.39 when spines first appear. Around the time of hatching, the hairlike appendages begin to disappear and spines become established. Reduction of spines occurs after hatching and continues to the adult stage. Possible influences of several external factors on neuronal maturation are discussed.

Notes: The female reproductive system of Sphaerodema rusticum consists of a pair of ovaries, two lateral oviducts, a median common oviduct, and a median spermatheca. Accessory glands are absent. Each ovary has five free ovarioles branching from the oviduct. Each ovariole consists of a terminal filament, germarium, vitellarium, brown mass, and an exceptionally long pedicel. The terminal filament consists of a central core, interstitial cells, and an outer sheath. In the germarium, which consists of trophic and prefollicular regions, the trophic region or nurse cell chamber is divided into four histologically differentiated zones, distinguished as zones I-IV. Nutritive cords, originating from the posterior end of the trophic core in zone IV extend centrally and join the developing oocytes in the prefollicular chamber and the vitellarium. The compact prefollicular tissue at the base of the trophic core gives rise to prefollicular cells which, after encircling the young oocytes, become modified into follicular epithelial cells, the interfollicular plug, and epithelial plug. The young oocytes descend into the vitellarium and gradually develop into mature oocytes. A compound corpus luteum is observed simultaneously in all the ovarioles of both ovaries after ovulation. Below the epithelial plug there is an accumulation of material, the “brown mass,” which develops cyclically in correlation with the ovulation cycle. Each pedicel stores five mature chorionated eggs ready for oviposition. The epithelium of the anterior region of the pedicel secretes a PAS-positive material. General morphology and histology of the subdivisions of the ovarioles are described.

Notes: The feeding mechanisms of two labrid fishes (Cheilinus chlorurus and C. diagrammus: Labridae: Perciformes) are modeled using four-bar linkage theory from mechanical engineering. The actions of the feeding mechanisms are simulated by a computer program that uses morphometric data to calculate the geometry of mechanism structure. The predictions of three different four-bar linkages regarding the kinematics of feeding are compared to the movements observed through hign speed (200 fps) cinematography. A previously unidentified four-bar chain was found to be an accurate model of the mechanism by which upper jaw protrusion, maxillary rotation, and gape increase occur in Cheilinus. This mechanism involves the anterior jaws including the mandible, maxilla, premaxilla, palatine, and suspensorium. The accuracy of two previously described four-bar linkages was also tested by comparison of model predictions and film results. The opercular linkage proposed by Anker ('74) as a mechanism of jaw depression via opercular levation was found to be a poor predictor of feeding movements. This four-bar chain involves the opercle, suspensorium, interopercle, and mandible. Muller ('87) proposed a mechanism of hyoid depression involving cranial elevation due to epaxial muscle contraction as input motion The links in this mechanism include the neurocranium and hyomandibula, hyoid, sternohyoideus muscle, and pectoral girdle. This model was an accurate predictor of hyoid depression in Cheilinus when simultaneous cranial elevation and sternohyoideus contraction were simulated. Quantitative kinematic models involve simplifying assumptions when applied to complex musculoskeletal systems, but such models have a wide range of applications to vertebrate functional morphology.

Notes: The morphology of both the main nasal cavity and the vomeronasal organ differs among species representing six families of caecilians. The main nasal cavity is either divided or undivided. The vomeronasal organ differs in position (mediolateral, lateral), size (large vomeronasal organ in the aquatic species), and shape (mediolateral extension, vomeronasal organ with a lateral rostral projection). The great amount of respiratory epithelium of the main nasal cavity, the large vomeronasal organ, and its extensive innervation in typhlonectids may reflect both phylogeny and habitat adaptation, for these taxa are secondarily aquatic or semiaquatic and have several concomitant morphological and physiological modifications. The vomeronasal organ is associated with the caecilian tentacle as the tentacular ducts open into it. This association is further evidence for the involvement of the caecilian tentacle in vomeronasal chemoperception and may represent the mechanism by which these animals smell though the main nasal cavity is closed during burrowing or swimming. Labelings of primary olfactory and vomeronasal projections by means of horseradish peroxidase reaction reveal that the pattern of vomeronasal projections is similar in Ichthyophis kohtaoensis, Dermophis mexicanus, and Typhlonectes natans, even though T. natans possess stronger vomeronasal projections relative to olfactory projections than I. kohtaoensis and D. mexicanus. However, there are differences with respect to the patterns of olfactory projections. The olfactory projection of I. kohtaoensis is characterized by many displaced glomeruli. T. natans has the smallest olfactory projection. The nervus terminalis is associated with the olfactory system as shown by selective labelings of olfactory projections.Six characters potentially useful for phylogenetic analysis emerge from this study of comparative morphology. The characters were subjected to analysis using PAUP to see (1) if any resolution occurred and (2) if any groups were distinguished, whether they corresponded to phylogenetic arrangements based on other morphological characters. The characters are too few to produce nested dichotomous sets for all cases, but they do support the two typhlonectid genera examined and Dermophis and Gymnopis as sister taxa discrete from other groups, and they show that species within genera cluster together.

Notes: A kidney from the budgerigar (budgie, parakeet; Melopsittacus undulatus) is composed of cortical reptilian-type nephrons (without loops of Henle) and mammalian-type nephrons (with loops) grouped together in medullary cones. The loop of the mammalian-type nephrons has a descending segment composed of thin and highly interdigitated cells. These thin limb cells have few mitochondria (15% of cell volume), undetectable Na+, K+-ATPase activity, and virtually no basolateral surface amplification. Prior to the hairpin turn, the descending limb thickens, but the cells continue to lack basolateral amplification. Cells just prior to and within the hairpin turn resemble cells of the entire ascending limb. These cells are thick (there is no thin ascending segment in the avian loop), with extensive infoldings of the basolateral membrane surrounding numerous mitochondria (45% of cell volume). The area of basolateral membrane is 25 times that of the apical membrane. The basolateral membrane (but not the apical membrane) is enriched in Na+, K+-ATPase activity. The structure of the avian mammalian-type nephron (as epitomized by the budgie nephron) and the fact that NaCl accounts for over 90% of the osmotic activity of avian urine leads to the conclusion that the countercurrent multiplier of the avian kidney functions by active NaCl transport from the entire ascending limb. No explanation is offered for the transport specializations found in the thick descending segment of the loop, just prior to the hairpin turn.

Notes: The cells-of-origin and the mode and site of termination of the interhemispheric connections passing through the anterior and posterior pallial commissures in the telencephalon of two lizards (Podarcis hispanica and Gallotia stehlinii) were investigated by studying the anterograde and retrograde transport of unilaterally injected horseradish peroxidase. The commissural projections arise mainly from pyramidal cells in the medial, dorsomedial, and dorsal cortices (medial subfield). Additionally some non-pyramidal neurons in the medial and dorsal cortices contribute to the commissural system. Medial cortex neurons project to the contralateral anterior septum through the anterior pallial commissure. The dorsomedial cortex projects contralaterally via the anterior pallial commissure to the dorsolateral septum and to the medial, dorsomedial, and dorsal cortices. The projection to the medial cortex terminates in two bands at the inner and outer border, respectively, of the cell layer; the projection to the dorsomedial and dorsal cortex ends in a zone in layer 1 which previously has been described to be Timm-negative, and in a diffuse band in the inner half of layer 3. The medial subfield of the dorsal cortex projects through the anterior pallial commissure to the dorsomedial and dorsal cortices with a similar pattern of termination to that found for the dorsomedial cortex. The posterior pallial commissure contains only the projections from the ventral cortex to its contralateral counterpart and to the ventral part of the caudal medial cortex. The similarities found between this commissural system and the mammalian hippocampal interhemispheric connections are discussed.

Notes: The distribution of structural and secretory glycoconjugates in the gastric region of metamorphosing Xenopus laevis was studied by the avidin-biotinperoxidase (ABC) histochemical staining method using seven lectins (concanavalin A, Con A; Dolichos biflorus agglutinin, DBA; peanut agglutinin, PNA; Ricinus communis agglutinin I, RCA-I; soybean agglutinin, SBA; Ulex europeus agglutinin I, UEA-I; and wheat germ agglutinin, WGA). Throughout the larval period to stage 60, the epithelium consisting of surface cells and gland cells was stained in various patterns with all lectins examined, whereas the thin layer of connective tissue was positive only for RCA.-I. At the beginning of metamorphic climax, the connective tissue became stained with Con A, SBA, and WGA, and its staining pattern varied with different lectins. The region just beneath the surface cells was strongly stained only with RCA-I. With the progression of development, both the epithelium and the connective tissue gradually changed their staining patterns. The surface cells, the gland cells, and the connective tissue conspicuously changed their staining patterns, respectively, for Con A and WGA; for Con A, PNA, RCA-I, SBA, and WGA; and for Con A, RCA -I, and WGA. At the completion of metamorphosis (stage 66), mucous neck cells became clearly idpntifiable in the epithelium, and their cytoplasm was strongly stained with DBA, PNA, RCA-I, and SBA. These results indicate that lectin histochemistry can provide good criteria for distinguishing among three epithelial cell types, namely, surface cells, gland cells, and mucous neck cells, and between adult and larval cells of each type.

Notes: Ultrastructural features of the ovotestes, spermatogenesis, and the mature sperm are described for three galeommatid bivalves, Divariscintilla yoyo, Divariscintilla troglodytes, and Scintilla sp., from stomatopod burrows in eastern Florida. All three species yielded similar results except with respect to mature sperm dimensions. The ovotestis contains three types of somatic cells within the testicular portion: flattened myoepithelial cells defining the outer acinal wall; underlying pleomorphic follicle cells containing abundant glycogen deposits; and scattered, amoeboid cells containing lysosomal-like inclusions which are closely associated with developing sperm. Early spermatogenesis is typical of that reported from other bivalves. In contrast, the late stages of spermiogenesis involve the migration and gradual rotation of the acrosomal vesicle, resulting in a mature acrosome tilted about 70° from the long axis of the cell. The mature sperm possesses an elongated, slightly curved nucleus; a subterminal, concave acrosome with a nipple-like central projection; five spherical mitochondria and two centnoles in the middlepiece; and a long flagellum. The rotational asymmetry and the presence of perimitochondrial glycogen deposits in these sperm are unusual in the Bivalvia and may be associated with fertilization specializations and larval brooding common among galeommatoideans.