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Notes: Neurohistochemical techniques were used to confirm morphologically the distribution of adrenergic and cholinergic nerves to the splenic microvasculature. The results form the basis of this report.Using these methods, adrenergic innervation was observed only in the adventitia of arteries and arterioles. No cholinergic innervation was found in this site. No adrenergic or cholinergic innervation could be demonstrated to the channels of the red pulp, venules or veins. These data provided morphological evidence that in the murine spleen only splenic arteries and arterioles are innervated; and these have only an adrenergic innervation.

Notes: The hypertrophic stage of development of the rat mandibular condyle was investigated in 16 and 26-day-old rats by electron microscopy. Interest was focused on the zone of mineralization and erosion. It was observed that capillaries invaded the lower level of the hypertrophic zone, without any previous chondroclastic resorption of calcified partitions. The partitions surrounding the hypertrophic chondrocytes were not mineralized around their entire circumference at the level of capillary penetration. The capillaries were accompanied by perivascular cells but these showed no similarities to chondroclasts. Multinucleated chondro- or osteoclasts were however present at a lower level of the subchondral area. It is suggested that there are no inherent differences with respect to the pattern of mineralization and erosion between the epiphyseal growth plate and the developing mandibular condyle.

Notes: The objective was to determine the distribution of growth hormone-release-inhibiting hormone (somatostatin) in the rat brain using the peroxidase-antiperoxidase immunocytochemical method with antisera prepared against unconjugated, synthetic somatostatin. Somatostatin occurred in low quantity in the organum vasculosum of the lamina terminalis. It was present throughout the full length of the median eminence and occupied the entire width between the tuberoinfundibular sulci. Most somatostatin was located in the dorsal portion of the external lamina, and the amount varied according to the mediolateral position. The bodies labeled for somatostatin were most often granules; occasionally they appeared as clusters of granules that seemed to be membrane-enclosed. Some of these bodies appeared to be portions of axons. Many of the larger bodies were arranged alongside tanycytes, but no label was distributed generally in tanycyte cytoplasm. Somatostatin was highly concentrated in the proximal one-quarter of the infundibular stem and appeared in lower concentration throughout the distal portion of the stem. It was absent from the pars nervosa and pars intermedia of the pituitary gland. The distribution of somatostatin in the median eminence differed considerably from that of gonadotropinreleasing hormone.Somatostatin was identified in the ventromedial and/or dorsomedial hypothalamic nuclei of only two animals. Here it was probably located in axons that terminated on neuronal cell bodies but also may have been present in a restricted portion of the perikaryonal cytoplasm.

Notes: A method is described for estimating the number of nuclei of different cell types in the sheep placenta. The method was applied to the study of six placentas of different ages and the results have been correlated with data on the chemical composition and the physiology of the ovine placenta. In agreement with studies of placental DNA content, there is no demonstrable increase in the total number of placental nuclei in the last two months of gestation. However, the number of fetal mesenchymal nuclei decreases and there is a concomitant increase of fetal endothelial nuclei from 80 days to term. These changes correlate with a decrease in volume of the core of the placental villi and increase in umbilical blood flow.

Notes: The ultrastructure and permeability of the apical junctions between epithelial cells of the distal nephron have been studied in rat kidney using a colloidal lanthanum tracer and uranyl acetate staining en bloc. The apical intercellular junctions of the macula densa and juxtaglomerular segment of the early distal convoluted tubule measure up to 0.5 μ in length and about 50 Å in width. Lanthanum permeates the occluding portion of these junctions in a discontinuous manner, defining a series of closely spaced and parallel lines of fusion that run in a direction perpendicular to the apical-basal axis of the tubular cells. The apical junctions of the remainder of the distal convoluted and cortical collecting tubules are impermeable to lanthanum. This distinctive apical tight junction can account for the greater permeability to ions of the early distal convoluted vs. late distal convoluted and cortical collecting tubules.

Notes: Human aorta was prepared for scanning electron microscopy using critical point drying. The aortic surface is lined by a continuous layer of squamous endothelial cells. The luminal surface of these cells contains many microvilli or microappendages and pinocytotic vesicles. In addition, at the periphery of each cell, larger marginal appendages or microfolds are present which allow the observer to see the cell boundaries clearly. The marginal folds are superficial to the cell junctions and could be observed without any staining procedures. These observations on human aorta are generally in agreement with our findings on the rabbit and guinea pig aorta and vena cava but are in contrast to others who have reported the presence of ridges, intercellular bridges, and “hair-like” processes on the endothelial surfaces.

Notes: When pancreases from fetal rats were transplanted beneath the kidney capsule of isologous normal adult recipients, continued growth and differentiation of the endocrine portion of the pancreas occurred. While limited amounts of acinar tissue were identifiable in the early transplant period (7 days), such cells were absent in long term transplants (14 and 21 days). In contrast, while few definitive islet beta cells were present at the time of transplantation, following 21 days at the kidney site large circumscribed islets comprised of heavily granulated beta cells in association with duct epithelial cells predominated. Mitotic figures were seen in both these cell populations. Total islet mass had increased over 20-fold during the transplantation period. Similar results were observed if fetal pancreases were grown in organ culture for ten days prior to transplantation. Continued islet and duct cell growth, as evidenced by mitotic figures and an increase in absolute islet cell mass was obtained in such cultured explants when transplanted to either isogenic or allogenic recipients.These observations support the hypothesis that fetal pancreas may be the best source of donor material for transplantation to diabetic recipients, in part, due to the continued growth and differentiation of the islet tissue during the transplantation period.

Notes: An extensive system of gap or nexus junctions has been found between astrocytic processes in the neuropil of the lateral hypothalamic area in the albino rat. These specialized interastrocytic junctions occur in regions of high synaptic density where neural processes are separated by the interconnected glial system. In this study, 90% of the gap junctions observed in the lateral hypothalamic neuropil are in the immediate proximity of synaptic terminals. The close morphological relationship of these glial gap junctions with synaptic contacts suggests that they may play a significant role in the process of synaptic transmission.

Notes: Incorporation of Fe55 in vivo was used for verifying on radioautographs the identity of chicken bone marrow cells that are in the process of heme syntheses. Under the experimental conditions all labeled cells may be considered to be linked with erythroid differentiation. They were classified into five maturational stages according to their morphology and capacity for DNA synthesis. Granulocytes were identified by the presence of specific granules. All mononuclear cells were classified as lymphocytes which had a pachychromatic nucleus, a high nucleocytoplasmic ratio, lacked the capacity for DNA synthesis, and resembled small lymphocytes of the bursa, spleen and bone marrow that bound, in vitro, anti-chicken gamma globulins labeled with I125. Radioautography with H3TdR was used to identify proliferating and non-proliferating members of each cell population.With the use of the morphological criteria thus established, reproducible differential counts could be performed on chicken bone marrow smears also in the absence of specific labeling. In normal, 3, 4, and 8 week old White Leghorn chicks, such counts revealed the bone marrow as a member of the lymphomyeloid complex preferentially adapted to the production and maturation of erythroid cells, while the reserve of granulocytes was found to be small compared to that of mammalian marrow. The percentage of lymphocytes appears to increase with age but, in contrast to rodent marrow, a proliferating precursor cell pool for lymphocytes could not be identified in chicken marrow up to the age of eight weeks.

Notes: Microridges (MR), also called microvillar ridges, microplicae and cytoplasmic folds, have been noted on many epithelial surfaces. Several functions have been proposed for these structures. In the present study we examine the mechanical role that microridges may play in holding mucus to the lumenal surface of the esophagus in the trout Salmo gairdneri. Our findings support the hypothesis that the microridges help hold a protective coat of mucus to the epithelium. In addition, the complex curved or whorled arrangement of microridges appears to facilitate the spread of mucus away from goblet cells.

Notes: The satellite cell population in skeletal muscles of ten normal subjects aged 7 to 73 years was quantitated by electron microscopy. In all 1,414 nuclei were assessed. Satellite cell nuclei comprised 4% (standard deviation 2%) of all nuclei within the fibre basal lamina. The lowest incidence (0.6%) was found in a 73-year-old man. The study confirms, that satellite cells are regularly present in human skeletal muscle.

Notes: Near the end of spermiogenesis, the late spermatids remain attached to the superficial layer of the seminiferous epithelium for an appreciable period of time (i.e., 3 to 4 days). The sickle-shaped heads of the spermatids are embedded in an apical process of Sertoli cell cytoplasm which is connected to the rest of the cell by a narrow stalk. In the concavity of the head several long (2-3 μm) and very narrow (50 nm) tubular projections of the spermatid's plasma membrane invaginate the Sertoli cell cytoplasm. These tubular processes terminate by a bulbous swelling which may measure up to 1 μm in diameter. Along the process the plasma membrane of the Sertoli cell is closely apposed to the spermatid's membrane, the intercellular space being only 6-8 nm wide. In the Sertoli cytoplasm immediately surrounding the tubular portion of the structure there is an accumulation of filamentous material, while next to the bulbous extremity there are, at a short distance, smooth surfaced cisternae of endoplasmic reticulum. The whole structure was referred to as a tubulobulbar complex. These complexes, of which there are up to 24 per spermatid, appear as these cells complete their migration toward the apex of the Sertoli cells. They disappear just before the release of the spermatids in the lumen of the seminiferous tubule as a result of the fragmentation of the spermatid's plasma membrane followed by a resorption of the Sertoli plasma membrane. Morphological evidence suggests that the tubulobulbar complexes serve as anchoring devices that retain the spermatids at the surface of the seminiferous epithelium while their dissolution contributes in part to the process of spermiation. Similar tubulobulbar complexes were also formed by the plasma membranes of two adjacent Sertoli cells close to the Sertoli-Sertoli tight junctions near the tubular limiting membrane.

Notes: The incorporation of 3H-tryptophan into the inner enamel epithelium of newborn mouse incisor tooth organs has been studied in situ by light and electron microscopic autoradiography to determine the sites and kinetics of biosynthesis, migration, and secretion of precursor enamel protein during newborn mouse incisor tooth formation. Maxillary and mandibular incisor tooth amelogenesis was studied 5, 30, 60, 120, 240 minutes and 24 hours following the intraperitoneal injection of 3H-tryptophan. By 5 minutes, 40% of the total silver grains associated with the secretory ameloblasts were localized over the rough endoplasmic reticulum and 50% of the silver grains were localized over the Golgi apparatus. By 30 minutes, silver grains were observed predominately over condensing vacuoles and secretory granules within the forming Tomes' processes, and were also localized over the extracellular “granular” pre-enamel matrix. The enamel proteins were synthesized on membrane-bound polysomes, transferred within the cisternae of the rough endoplasmic reticulum and then accumulated in the inner saccules of the Golgi apparatus. The enamel proteins were then packaged in condensing vacuoles which subsequently became secretory granules which migrated to the lateral and apical secretory regions of the forming Tomes' processes. It was concluded from these in vivo studies that enamel proteins were synthesized and subsequently secreted within 30 minutes. The initially secreted precursor enamel protein was localized over a material which demonstrated a granular or stippled ultrastructure. The labeled protein then was localized over the amorphous enamel matrix per se which contained the forming calcium hydroxyapatite crystals. We assumed, therefore, that there are two different ultrastructural forms of 3H-tryptophan containing extracellular enamel proteins and suggest that the granular or “stippled” form represents newly secreted precursor enamel protein.

Notes: Oocyte growth in relation to follicular growth in marsupials conforms to the uniform biphasic pattern characteristic of eutherian mammals. However, the oocyte of the marsupial is larger and completes growth in a correspondingly larger follicle. At the completion of oocyte growth the follicle, like that of eutherians, shows no appreciable accumulation of follicular fluid. The ratio of oocyte/follicular diameter in marsupials at this stage is 0.59, a value close to that of 0.55 for eutherian mammals. The oocyte nucleus of the marsupial is also significantly larger than that of the eutherian, but the oocyte nucleolus is approximately the same diameter.By contrast, the growth of the oocyte in relation to the follicle in monotremes is linear throughout the whole period of follicular growth. However, like eutherians and marsupials oocyte nuclear and oocyte nucleolar growth are arrested in the early stages of follicular growth. The diameters of the oocyte nucleus and nucleolus at the completion of growth are approximately twice those found in marsupials.It is concluded that the growth patterns of the oocyte nucleus and nucleolus are common to all mammals but that the biphasic pattern of oocyte and follicular growth which is characteristic of eutherian and marsupial ovaries is not found in the monotremes.

Notes: The exfoliating epithelial surface of the ectocervix (pars vaginalis) and the vagina from normal mice was studied by scanning electron microscopy during the four main phases of the ovulatory cycle, and after estrogen administration. At proestrus, the epithelial surface usually presented cobblestonelike structures furnished with thin microvilli; at estrus, large, flat mosaics covered by long, convoluted microrugae; at metestrus, the presenting cellular surfaces were of intermediate size (i.e., between cobblestone and mosaics) having fragmented microrugae; and at diestrus, by mosaics or intermediate structures covered by microvilli. Estrogen administration to intact and oophorectomized animals resulted in flat mosaics and large numbers of loosely attached cells with rolled-up borders which permitted the observation of the basal surface of the cells. In those animals the apical aspect of the cells which lined the lumen showed convoluted microrugae with circular configuration, while the basal aspect of such cells had club-tipped microvilli. The findings were correlated with post-scanning histological examination in all animals.

Notes: Studies on the growth sequence of follicles in mature mice four to six months old were conducted by giving three injections of 16 μCi tritiated thymidine (3H-T) / gm body weight at four hour intervals over a period of eight hours. Subsequent radioautographic analysis on ovaries obtained 1, 3, 5, 9 and 17 days after the last injection showed an overall follicle growth time of 17 to 19 days. The duration of the different follicle stage times were also estimated from radioautographic data. Intense 3H-T incorporation was noted in the developing antra and zona pellucida of follicles of animals sacrificed one and three days after treatment suggesting that these areas serve as precursor storage sites during development. Oocytes of follicles in early antrum formation also showed juxtanucleolar 3H-T incorporation concomitantly with rapid and massive oocyte and follicular growth. The findings further indicate that the earliest follicular cells surrounding the oocyte originate at some distance from the developing primordial follicle.

Notes: Tensile breaking load, strength, strain, modulus of elasticity and density plus the histological structure at the fracture site, were determined for 207 standardized specimens of cortical bone from the embalmed femur, tibia, and fibula of 17 men from 36 to 75 years of age. The men were divided into a younger group (41.5 years old-avg) and an older group (71 years old-avg). Specimens from younger men had a greater average breaking load, strength, strain, modulus and density than those from older men. The percentage of spaces in the break area was greater in specimens from older men, but specimens from younger men had a slightly greater percentage of osteons, osteon fragments, and interstitial lamellae. The number of osteons/mm2 and of osteon fragments/mm2 was greater in specimens from older men but the average area/osteon and area/osteon fragment was greater in specimens from younger men. Thus, there are quantitative and qualitative differences in the histological structure of bone from younger and older men. Differences in the tensile properties of bone from younger and older men can be explained by histological differences in the bone.

Notes: Young adult male rats were treated with daily injections of 10 mg of cyproterone acetate for periods up to 16 weeks. Samples of the ventral prostate and the seminal vesicle were studied with the light and electron microscopes. Alterations visible with the light microscope included decreases in cell size, cytoplasmic basophila, the size of the nucleolus, and the amount of luminal secretory material. Ultrastructural changes in the epithelium of both glands involved mainly the organelles that participate in the formation of secretions. Large declines were observed in the abundance of rough endoplasmic reticulum, size of the Golgi apparatus, and number of secretory vacuoles. Lipid droplets accumulated in the seminal vesicle epithelium, and lysosomes were numerous in both glands. Changes were first observed microscopically in the seminal vesicle after one week and in the prostate after two or three weeks. Maximal development of the alterations occurred after treatment for approximately eight weeks.

Notes: Earlier work has suggested that terminal nerve fibers in the periphery of the dental pulp are linked to one another and to odontoblasts by gap junctions although positive identification of the junctions was lacking. In this study, the lanthanum hydroxide tracer technique has been used to demonstrate that the junctions in this area do have the characteristic features of gap junctions, a wide dark intermediate line in transverse section, stippling in oblique section and a globular arrangement of sub-units in tangential section. Other studies have shown these junctions to be of low electrical resistance although the functional importance of this characteristic in this situation has not been determined.

Notes: Using a combination of single isotope and double isotope autoradiography after injection of 3H-thymidine and of 3H- + 14C-thymidine, respectively, the cell cycle of normal and injured lens epithelial cells in the mouse was determined. Progenitor cells in the peripheral region of normal lens epithelium were found to traverse the replicative cycle in the same amount of time as the injury-stimulated cells in both peripheral and central (wound) regions. The individual phases were also the same length, except for a slight shortening of G2 in the injured epithelium. The durations of the phases were: S: 11-12 hours; G2: 1.5-2 hours; M: 4.2-5 hours; G1 (derived): 38-44 hours; and total cycle: 55-61 hours. Two findings were significant in view of previous observations that injury to mouse lens seldom induces lens opacity. First, while the cell cycle duration was not affected by injury, a burst of proliferation of potentially active cells ensued. And, secondly, this burst of proliferation involved only one cell cycle. Only a small number of cells, equivalent to the normal progenitor compartment, continued into a second replicative cycle. The wound was healed, therefore, mainly by one division cycle involving a large number of cells.

Notes: The contribution of progenitor cells in the normal peripheral region of mouse lens epithelium to healing of a centrally located wound was studied using an autoradiographic technique designed to follow migrating 3H-thymidine labeled cells. It was found that few progenitor cells labeled before or at the time of injury migrate toward the wound and hence contribute little to the repair of the wound. Moreover, the central injury did not delay or reverse the normal migration of the labeled peripheral cells in the opposite direction into the bow. It is concluded from this and other studies that a central injury to the mouse lens is healed mainly by numerous local cells which are stimulated for the most part to traverse one replicative cycle of normal duration, and that the injury does not affect formation of new lens fibers. These findings are thought to be related to the observed low incidence of traumatic cataract in the mouse.

Notes: At the close of a uterine cycle, the remodelling of the endometrial stroma of the North American opossum involves removal of extracellular material by macrophages. This study provides cytochemical and ultrastructural evidence which indicates that the laden macrophages are eliminated from the endometrium through emigration across the glandular and luminal epithelia. During diestrus or the early postpartum period, the abundant uterine glands relinquish their secretory function to acquire a transient function in the transportation of emigrating stromal cells.During the first three postpartum days endometrial regression in the stroma is marked by sudden appearance of monocytes, macrophages, lymphocytes, and plasma cells. Ultrastructural and cytochemical evidence indicates that the macrophages engulf the extracellular macromolecular material which, in the opossum, consists primarily of ground substance. Macrophages filled with ingested extracellular material aggregate beneath the glandular and luminal epithelia, where they acquire an extracellular coat that resembles the material of the basal lamina elsewhere. A fibroblast-like cell closely invests the macrophage at the time the extracellular material appears. Simultaneously, the secretory glandular epithelium is being converted to a highly ciliated one. Macrophages, often accompanied by lymphocytes, acquire intraepithelial positions in the glands. From here these stromal cells gain entrance to the glandular lumens. At this time the luminal contents are rich in acid phosphatase activity which most likely reflects the high lysosomal content of the emigrating macrophages. Evidence suggests that these intraluminal macrophages and lymphocytes are swept, by the recently differentiated ciliary lining, toward the glandular orifices and into the uterine cavity. It is hypothesized that this cyclic appearance and transepithelial elimination of macrophages is a cellular mechanism for removing large amounts of extracellular material without disruption of the endometrium.

Notes: Multiply ciliated follicular cells have been observed in developing thyroid glands in dogs. These cells are prominent three weeks prior to the expected date of birth but become progressively less common as development proceeds. After birth only rare isolated cilia remain. No evidence of removal of cilia by autophagy or loss into colloid was found. It is concluded that cilia are probably no longer formed by mature cells and that those already present are diluted out by progressive division of mature follicular cells.

Notes: The present study was initiated to determine whether specific hormones would influence adenylate cyclase activity within the maxillary-palatal complex during formation of the hamster secondary palate. Stages from initial appearance of the palatal processes to shortly after birth were studied. Highest basal adenylate cyclase activities occurred during the earliest periods of palate development. This basal enzyme activity began to diminish as palatal fusion occurred and remained lowered until birth. Activation of adenylate cyclase by fluoride was maximal at concentrations of 5-10 mM, and was observed throughout the span of palatal development. Fluoride activation of adenylate cyclase was greatest prior to fusion of the palatal processes, then decreased until birth when a slightly increased enzymatic stimulation was seen. Norepinephrine and epinephrine were the catecholamines most capable of inducing increased activation of adenylate cyclase at most periods of palatal growth. Increased enzyme activity in the presence of norepinephrine was more susceptible to antagonism by the beta adrenergic agent, propranolol, than to the alpha adrenergic agent, phentolamine. The remaining catecholamines, namely isoproterenol and dopamine, displayed a lesser ability to activate the enzyme, and adenylate cyclase was not equally responsive to these catecholamines at identical developmental stages. Other hormones, i.e., histamine, serotonin, thyrotropin, growth hormone, thyroxine and glucagon were generally ineffective in activating the enzyme. Phosphodiesterase activity was not detected until shortly before birth.

Notes: In most marsupials, placentation involves only the yolk sac; however, in the bandicoot family, Peramelidae, a functional chorioallantoic placentation develops in addition (Hill, 1895, 1897, 1900; Flynn, ′22, ′23). This duality is viewed as having evolutionary significance because most eutheria have both placentae. Furthermore, the bandicoot trophoblast was reported to vanish from the chorioallantoic site in late gestation (Hill, 1897; Flynn, ′23); whereas, the eutherian trophoblast is identifiable throughout later pregnancy and may act as an immunological barrier between maternal and fetal genotypes (Kirby, ′68). Thus we have re-examined this singular chorioallantoic placenta of the bandicoot in plastic sections with light and electron microscopy.A distinctive feature of bandicoot placentation is the transformation of the uterine simple columnar luminal epithelium into a highly vascular lining composed almost entirely of discrete syncytial masses (homokaryons). Endometrial blood vessels penetrate among the homokaryons to create a rich network of large diameter capillaries at extremely superficial locations near the maternal surface.In the chorioallantoic placenta (7 mm to 10-11 mm crown-rump embryos) the microvillous surface of the maternal homokaryons interdigitates with the microvillous border of the fetal trophoblast with desmosomal interaction. This trophoblast consists of a single layer of tall columnar undifferentiated cells rich in ribosomes-polysomes, poor in cytoplasmic membranes, and with large nuclei that have distinct clumps of heterochromatin and conspicuous nucleoli. It is thus remarkable that these undifferentiated cells disappear as a recognizable layer later in gestation (12 mm crown-rump embryos).Flynn's hypothesis that the trophoblastic cells disapppear by fusing with maternal syncytia gains support from the existence of two populations of nuclei in the syncytial masses only at the chorioallantoic site. One population is comparable to that occurring in the homokaryons of the yolk sac placenta, i.e., pale staining nuclei with little heterochromatin and small peripheral nucleoli. However, the other nuclei resemble those of the trophoblast cells. Since the trophoblastic cells before their disappearance as a layer possess properties associated with potential for further differentiation, the possibility of fusion between the maternal homokaryons and the fetal trophoblastic cells to form heterokaryons composed of two genotypes merits further consideration.The disappearance of the trophoblastic layer and the superficial positioning of the maternal capillaries bring the maternal and fetal bloodstreams into closest proximity near term (12 mm crown-rump embryo). The thinnest parts of the barrier consist of delicate cytoplasmic extensions from the syncytial masses (that may be maternal in origin or jointly maternal and fetal) and a layer of maternal stroma intervening between the maternal and fetal endothelia. Thus the chorioallantoic placental barrier of the marsupial bandicoot is unlike any thus far described for eutherian mammals.

Notes: 9+0 cilia associated with diplosomes were observed in myoepithelial cells of the ductal system in the undiseased, resting, adult female breast. Ciliary diameter was 250 nm, and the longest found was 830 nm. The centrioles comprising the diplosome were 500 nm in length and 160 to 200 nm in diameter. Satellite bodies were associated with the diplosome but there was no rootlet system. Evidence suggested that all myoepithelial cells bear a cilium. The possible functions of the cilia in a sensory role as chemo- or mechanoreceptors related to myoepithelial and epithelial activities are discussed.

Notes: The neurothelium obtained from human biopsy specimens is a very thin cellular layer located between the dura mater and the arachnoidea. It is a pluri-stratified squamous layer which has some epithelial features (desmosomes and tonofilaments), and lacks connective tissue fibers in the extracellular spaces. Usually there is increased nuclear and cytoplasmic density and enlarged extracellular spaces from the deepest to the most superficial layer.Since neurothelium is fragile and easily torn apart, it probably occupies the position considered to be the subdural space.

Notes: The ligamentous anatomy of the wrist was analyzed by studying (1) 28 dissected human wrists, (2) and by examination of 19 wrist injuries created mechanically. Six volar intracapsular ligaments are described, two stabilizing the distal carpal row, three stabilizing the proximal carpal row and one stabilizing the proximal scaphoid pole. More specific terminology for these ligaments is proposed. An explanation of how these ligaments participate in the mechanical functioning of the carpus is advanced. Mechanically created wrist injuries had ruptures of specific ligaments as described augmenting a need for a more specific terminology. Suspected mechanisms of injury in perilunate and lunate dislocations are mentioned based upon the functional understanding of the intracapsular ligaments.

Notes: The appearance and development of the primary and secondary sympathetic trunks in staged chick embryos was studied using the Falck-Owman histochemical method for the demonstration of primary monoamines. The earliest appearance of catecholamine (stage 20) was in individual fluorescent cells located in the region of the dorsal root ganglia about two stages prior to the formation of primary trunk aggregates. These cells are believed to be sympathetic precursor cells and correspond to formaldehyde-induced fluorescent cells observed in recent explantation experiments. Aggregates of fluorescent cells had formed bilaterally dorsolateral to the aorta at stage 22. These aggregates became continuous to form primary trunks by stage 24. The secondary sympathetic trunks were first seen in stage 25 and appeared to form at least partially by dorsal migration of cells from the primary trunks. Fluorescent cell processes were first observed at this stage. Secondary trunk formation was essentially complete by stage 28, and the primary trunks had become small and discontinuous. Definite rami communicantes could be observed by the early part of stage 28 in silver preparations. The significance of the development of two successive trunks in avians is discussed.

Notes: The hamster accessory optic fiber system has been investigated with the use of de Olmos-Ingram and Fink-Heimer silver methods following the production of unilateral ocular enucleation. It was found that this fiber system consists of both crossed and uncrossed inferior and superior fasciculi. The fibers of the inferior fasciculus (anterior accessory optic tract) run along the medial edge of the cerebral peduncle and terminate within the medial terminal nucleus of the accessory optic system. The fibers of the superior fasciculus (posterior accessory optic tract) leave the main optic tract, pass superficially over the medial geniculate nucleus and the cerebral peduncle; they synapse within the dorsal, the lateral and the medial terminal accessory optic nuclei. The presence of a retinohypothalamic tract could not be confirmed.

Notes: Pregnant Wistar rats injected intraperitoneally on gestational day 12 with single doses (100-1,000 mg/kg) or 600 mg/kg of 4(5)-(3,3-dimethyl-1-triazeno)imidazole-4-carboxamide (DIC) were autopsied on day 21 (100-1,000 mg/kg) or at 24-hour intervals on days 13-20 (600 mg/kg). Controls received CMC on the same schedule. All fetuses were weighed and examined for urogenital system (UGS) malformations. Those given 600 mg/kg were also studied histologically.DIC produced significant growth retardation at all doses on day 21 (18-72%). UGS malformations occurred in 27-67% of the fetuses at 200-400 mg/kg and in 100% of those given 600 mg/kg or more of DIC. Abnormalities included renal growth inhibition, fusion, ectopia, and ureteropelvic dilatation. At 600 mg/kg renal and body weights were reduced 40 and 55%, respectively. Ureteropelvic dilatation was common, and cortical glomeruli, nephric collecting tubules, and papillae were retarded in development. The juxtamedullary glomeruli were well developed. Proximal nephric tubular mitotic activity was 85% greater than in control animals (day 17).On the basis of pertinent morphological and physiological data, it is postulated that the dilated upper urinary tracts represent functional hydronephrosis incident to severe renal retardation and its resultant compensatory response.

Notes: The nuclear fibrous lamina, a structure of unknown function, is situated at the nuclear periphery directly apposed to the inner nuclear membrane. Although it has been well described in invertebrate and many non-human vertebrate cells, its occurrence in human tissue has not been emphasized. This survey of a large variety of human cells has documented the lamina to be present, although of varying thickness, in a large number of cell types. It is consistently thickest in mesenchymal or mesenchymally derived cells and its apperance and thickness is not correlated with pathologic alterations.

Notes: Microfilaments at the junctional specializations between adjacent Sertoli cells and between the Sertoli cell and the late spermatid of the mouse and swine testes bind HMM and form arrowhead complexes with a periodicity of about 35 nm. The arrowhead formation is inhibited when the tissues are treated with HMM in the presence of ATP. These observations show that the microfilaments are actin-like in nature. The functional significance of these filaments in the Sertoli cell is discussed.

Notes: Scanning electron microscopy (SEM) is a potent tool that is especially valuable in interpreting the three-dimensional relationships of cells within tissues. This type of information is obtainable from thin sections in transmission electron microscopy (TEM) only by reconstructions of serial sections. The arrangement of the interstitial cells of the testis in relation to the capillaries and lymphatic channels, in particular, is easier to visualize in SEM than in TEM. Cytoplasmic constituents, as well as cell surface modifications, are demonstrable by this technique. The presence of droplets, presumably lipid droplets, both within and on the Leydig cells and the lymphatic endothelial cells, is quite evident. Other cytoplasmic structures are also apparent. For example, the possible functional significance of “openings” that are seen by SEM on the septa that surround lipid droplets is discussed relative to the appearance of the same area as seen in thin sections or in freeze-fracture replicas. SEM should become a very useful method for studying cytological and morphological alterations that occur in testicular tissue that is subjected to physical or chemical manipulation.

Notes: Pineal complexes in 85 species of lizards examined comprised seven morphological types. Members of the same family do not necessarily have the same pineal complex type. “Regressive” parietal eyes were not common except in certain arboreal lizards, primarily from the family Chameleontidae. The parietal eye is often retained in burrowing lizards, presumably because these animals are occasionally exposed to light and the parietal eye is a more suitable photoreceptor for a burrower than are lateral eyes. The pineal of certain lizards possesses a finger-like projection that extends toward the parietal eye. This extension, along with pineal wall convolutions, results in more photoreceptor cells oriented for maximal absorption of light. It is rare to find convolutions and an extension in the same pineal. Cartilage deposits and blood sinuses may modify the intensity and wavelength of light reaching the pineal. These observations suggest that the intracranial pineal of lizards is a more important photoreceptor than was previously realized, a situation that may be a factor in the occasional “failure” of parietalectomy experiments.

Notes: This study covers the period from the fortieth day after ovulation to the thirty-eighth day after birth. In addition, lungs of two mature monkeys have been analyzed. Birth occurs about the one hundred sixty-eighth day (24 weeks after ovulation). By the fortieth day the patterns of the bronchopulmonary segments are well established. These are the tools of the investigator. The curve of growth for the first 60 days parallels that of the rhesus monkey but is steeper than the human curve. At 63 days (9 weeks) pairs of rosette-like clusters of epithelial buds (future alveolar ducts) have appeared on peripheral branches of the segmental bronchi and continue to grow in size until the twelfth week. Then they resemble the paired rosettes of human acini of the seventeenth postovulatory week (Boyden, ′74) and probably represent the primitive unit from which the human was derived. In both man and monkey, canalization of the rosettes starts at about the same relative time (the 17th and 12th week, respectively) and spreads centralwards from the rosettes along the bronchioles of each segmental bronchus. In man, canalization forms acini (alveolar ducts and respiratory bronchioles). In the monkey, due to the monopodial pattern of branching, it produces racemes of alveolar ducts, each raceme ending basally in a pseudorespiratory bronchiole (127th day). By the one hundred fortieth day, enough surfactant has been secreted to support a premature infant. At one hundred fifty days, 11 generations of bronchi, followed by 16 of pseudorespiratory bronchioles, are present in the medial basal segment, followed in turn by 19 generations of prealveolar ducts (the latter provided with delicate spiral muscles) each giving off alternate branches of alveolar ducts. After one hundred fifty days of gestation, generations of bronchi, pseudorespiratory bronchioles, prealveolar ducts and cartilages (15) remain virtually constant for at least 38 days after birth. By the adult stage the number of generations of pseudorespiratory bronchioles has been reduced to three and the prealveolar ducts increased (at the former's expense) to 27. This study is concluded with a comparison of the lungs of the newborn monkey with those of the human/neonate and puppy.

Notes: The pars intermedia of non-obese pigmented rats of the Zucker strain is described using transmission electron microscopy. Secretory cells appear similar to those described elsewhere in albino rodent hypophyses. Cells which may be equivalent to ACTH cells of non-pigmented mice and rats are observed near theborder of the pars distalis. Melanocytes are observed to surround lobules of the pars intermedia and to penetrate among secretory cells. In addition, large granules which appear to be melanin are present within some pars intermedia cells. Portions of some cells near melanin or melanin-like granules exhibit expanded rough endoplasmic reticulum and Golgi zones containing dense granules. Observations are discussed in relation to origin of the melanin, and its possible significance near cells known to produce melanocyte-stimulating hormone (MSH).

Notes: The relationship between developing spermatids and Sertoli cell junctional specializations was studied with the electron microscope during spermiogenesis and at spermiation. At stage I of the seminiferous cycle, the newly formed spermatids are found in apposition to junctional specializations at the lateral surfaces of the Sertoli cell. Visualization of the junctional site of this early stage appears to be dependent on orientation and plane of section. As differentiation proceeds, the spermatids elongate and come to lie within deep recesses of the Sertoli cell. At this time the junctional specialization is limited to the acrosomal portion of the spermatid. During the maturation phase, the spermatids, while maintaining the same relationship to the junctional specialization, approach the lumen. When stage VIII of the cycle is reached, the stage in which spermiation occurs, the spermatids are at the luminal surface. The relationship of the spermatid head to the junctional specializations is quite variable during this stage. Some spermatids are observed still attached to the Sertoli cell at the junctional site, while others are found completely or partially surrounded by Sertoli cytoplasm, but with no evidence of the normally interposed junctional specialization. Yet, in other instances, the spermatids are observed in a position slightly removed from the junctional site. Also evident are profiles of junctional specializations at a free surface of the Sertoli cell, there being no attached spermatid. In some instances the junctional specializations appeared in apposition to a residual body. In the case of the free surface profiles, the junctional specialization at times lined an empty cleft or crypt-like recess, giving the impression that the spermatid head had just been dislodged from the junctional contact site. The findings indicate that the spermatid is in contact with a junctional specialization from its initial appearance and remains so until spermiation is initiated. It is postulated that spermiation is initiated through a physiological change in the junctional specialization resulting in loss of adhesion and consequent release of the sperm head from its attachment site. A similar mechanism is proposed in relation to the inter-Sertoli junctional complex to account for the means by which the spermatocytes cross this barrier to reach the adluminal compartment of the seminiferous epithelium.

Notes: The macroscopic anatomy of the brain of the American Plains Buffalo (Bison bison bison) is detailed herein for the first time. Many features vary significantly from most previously described ungulate species including its large size, fissural pattern of the cerebral hemispheres and well developed insular cortex. The midbrain, particularly the tectum, is outstanding: the superior colliculi appear to be the largest reported for any mammal and occupy most of the dorsal midbrain by surface area and volume estimates. The cerebellum is small relative to the total brain volume, a peculiarity reported elsewhere for other ungulates. Morphometric criteria further indicate the brain of bison is highly specialized. Drawing from various lines of evidence, it is suggested that the anatomical uniqueness of the present day bison brain may represent a vestige of the past, remaining largely unchanged over a broad expanse of geologic time.

Notes: The surface coat of syncytial trophoblast from term human placentas was studied using cytochemical methods (colloidal iron, alcian bluelanthanum nitrate, dialyzed iron) in coordination with tissue enzyme digestions (trypsin, neuraminidase) and sialic acid analyses. The presence of at least two highly acidic anionic components that contribute significantly to the surface negativity of trophoblast has been demonstrated. The first of these, sialic acid, was removed with neuraminidase. Tissue digestion with this glycosidase was accompanied by a decrease in trophoblast surface staining with colloidal iron, a decrease in tissue sialic acid, and an increase in the concentration of sialic acid in the incubating medium. Results from methylation experiments were consistent with the presence of sialic acid. The second anionic component(s) was identified by removal with trypsin of a glycocalyx constituent that stained with both colloidal iron and lanthanum. After trypsinization, tissue sialic acid levels were not significantly different from control values, and no detectable sialic acid was present in the incubating medium. The identity of this anionic component has not been established. Both sialic acid and nonsialic acid acidic components are distributed in higher density on membrane of microvilli than on intermicrovillous surface membrane. In addition, the sialic acid moieties appear to be clustered in the glycocalyx.

Notes: Explanted blastoderms of freshly laid chicken eggs expand their area during the first 44-45 hours of incubation by a factor of at least 11 if they are placed with the epiblast on the inner surface of explanted fresh chick vitelline membrane and provided with chick egg extract. This expansion is due essentially to the spreading of the yolk sac-serosal membrane. On turkey and duck membrane the expansion factor is about 6 and 3.8 respectively under otherwise identical conditions, but 1.9 only on a semisolid nutrient agar plate. Only the inner surface of the vitelline membrane has this growth-promoting potential, which markedly and progressively declines during incubation in ovo because of systemic factors rather than because of a direct influence by the outgrowing yolk sac-serosal membrane. Trypsinization of fresh chick vitelline membrane (1% trypsin 3 hours) reduces the growth-promoting potential to about 40% of its normal strength. The outgrowth of the extraembryonic tissues on vitelline membrane is better supported in the presence of a species' own egg extract than by extract from another species.

Notes: Rat testes fixed by vascular perfusion were examined by scanning and transmission electron microscopy. This revealed a complex organization of the interstitial tissue and an extensive “testicular lymphatic space” composed of continuous “peritubular lymphatic sinusoids.” No cellular connections exist between the interstitium and the tubules. Each sinusoid completely surrounds an individual seminiferous tubule, and adjacent sinusoids communicate freely through fenestrae in the interstitium. Thus, material must enter the lymph to gain access to the tubules. The sinusoids are delimited by separate endothelial cell layers over the tubules and interstitium. The layer over the interstitium is discontinuous in specific areas according to the geometry of the tubule packing. Interstitial tissue located in the flattened biconcave interstices between two adjacent tubules is consistently covered with endothelial cells, while that located in the open, triangular interstices between three or more tubules consistently lacks an endothelium. Most of the Leydig cells are located in the “open interstitium” and are directly exposed to the lymph. Physiological implications of the “testicular lymphatic space” and additional features of the interstitium are discussed.

Notes: Pieces of rat and human gastric mucosa were fixed in glutaraldehyde and paraformaldehyde and were processed by the methods of Wach-stein-Meisel and of Ernst for the cytochemical demonstration of adenosine triphosphatase (ATPase) and p-nitrophenylphosphatase (NPPase) respectively. Electron microscopic examination of reacted sections revealed the ATPase reaction to be localized to the plasmalemma and to mitochondrial cristae and the NPPase reaction to the plasmalemma. The tubulovesicles were unreactive. By thus demonstrating a cytochemical difference between the plasmalemma and tubulovesicle membrane, this study suggests that the tubulovesicles do not represent an extension of or reserve of mature plasmalemma. If the tubulovesicles represent a reserve of membrane that can be rapidly utilized to expand the plasmalemma, then these studies suggest that additional membrane constituents are added, activated, or unmasked in the process.

Notes: The effects of colchicine and vinblastine on tanycytes of the rat median eminence have been studied using the electron microscope. Colchicine and vinblastine were administered by intraventricular cannulation into the third ventricle and demonstrated distinct morphological effects on tanycyte microtubules. Highest drug doses administered were 50.0 μg in 5.0 μl of saline initially, followed by an additional 50.0 μg of these agents infused in a volume of 13.0 μl saline over an hour. Colchicine treatment resulted in the formation of large crystalloids within tanycytes, coincident with the disappearance of microtubules, all along the ventricular surface. Nonetheless the tanycytes appeared able to maintain a continuous tight lining, as in controls, although some crystalloids were observed beneath the ventricular surface, either within tanycyte processes or within neuronal processes. Vinblastine treatment also resulted in the formation of identical crystalloids but at highest doses thoroughly destroyed the tanycyte lining of the median eminence and exposed cellular elements below the surface to the drug. Portal capillaries approaching the ventricular surface were generally free of any signs of drug response to either colchicine or vinblastine.

Notes: A circadian rhythm in the incorporation of tritiated thymidine into DNA of the mouse thymus has been described. This same rhythm in animals bearing an 8-day Ehrlich ascites tumor (EAT) was altered in its phasing, wave-form and over-all 24-hour mean; the most striking alteration seen in four out of five studies was a reduction in the over-all DNA synthesis rate in tumor-bearing animals.Mice subjected to an altered light-dark cycle did not show a complete phase shift in their thymus DNA rhythm within two weeks, but during this time the rhythm in the mitotic index of their corneal epithelium completely phase shifted; the presence of the tumor had no effect on this latter rhythm. In mice subjected to a daily 4-hour restricted feeding schedule, the rhythm in DNA synthesis in the thymus became synchronized in that the peak occurred toward the end of the 4-hour restricted feeding span in both tumor and non-tumor-bearing animals. The rhythm in the mitotic index in the cornea was not phase shifted or altered by the restricted meal timing.The DNA synthesis rhythm in the normal thymus demonstrated a phasing similar to (1) the rhythm in the length of survival time in untreated mice challenged with EAT and (2) the rhythm in DNA synthesis in the normal spleen.

Notes: The frequency of labeled mitoses method (FLM) was used to study the cell kinetics in the stratified squamous epithelium of the mouse esophagus and tongue. FLMs were generated by injecting tritiated thymidine (TdR) at two different phases of the mouse circadian system: TdR was injected into one group of mice at 0900 and into a different, second group of mice at 2100. Three variables were monitored for each group; (1) the FLM, (2) the mitotic index and (3) the grain count over the labeled mitotic figures. In both the esophagus and the tongue there was a circadian rhythm in the mitotic index with the peak occurring during the first half of the diurnal phase and the trough occurring during the first half of the nocturnal phase. The FLM curves from each group revealed the following data: TextTransit time in G2+1/2M (TG2+1/2M)Transit time in S phase (TS)hourshoursEsophagus  -  TdR at 09002.55.3Esophagus  -  TdR at 21004.53.5Tongue  -  TdR at 09004.25.1Tongue  -  TdR at 21004.25.5Second waves of labeled mitoses were seen in the esophagus and tongue when TdR was injected at 0900. No second wave of labeled mitoses occurred in either the esophagus or the tongue when TdR was injected at 2100. For the esophagus the grain counts of the labeled mitotic figures in each group indicated that the rate of DNA synthesis was greater during late S phase, whether the TdR was injected at 0900 or 2100. During most of the S phase the rate of DNA synthesis was significantly greater in the TdR at 2100 group, which is the time of the circadian system when the esophagus demonstrated a relatively short Ts. In the epithelium of the tongue there were a few statistical differences in the rate of DNA synthesis between the groups and during the duration of the S phase within either group, but these data were difficult to interpret.The data demonstrate the influence of the circadian system in cell kinetic work and in the interpretation of data obtained with the FLM in renewing populations of cells which are not totally asynchronous.

Notes: The in vitro incorporation of 3H-5-uridine into the germinal and somatic cells of the larval ovary of Xenopus laevis has been studied using both light and electron microscope autoradiography. Incubation for only one hour in the presence of precursor revealed that the follicle cells are highly active in rRNA synthesis, whereas substantial oogonial nucleolar labeling was not detected for several hours. Semi-quantitative analysis of high-resolution autoradiograms indicated that the density of silver grains associated with “nuage” in oogonia was almost 4-fold greater than the surrounding cytoplasm. This strongly suggests that a significant amount of RNA is associated with “nuage” at this stage of Xenopus oogenesis, in addition to its well documented protein composition. “Pulse-chase” experiments further suggest that the nuage-associated RNA is stable for at least 24 hours. These results are discussed (within the limitations imposed by the methodology) both in relation to other studies on the composition of nuage in a wide variety of germinal cell types and in the light of growing speculation that nuage and germinal granules are synonymous.

Notes: Submandibular glands from 17-day-old rat fetuses were maintained in organ culture for five days in a medium consisting of Eagle's MEM (87%), horse serum (10%), and chick embryo extract (3%). Each day of the culture period explants were incubated for the demonstration of peroxidase activity and processed for light and electron microscopic observations. In some experiments cultures were exposed to 3H-thymidine one hour prior to fixation and incubation for the demonstration of peroxidase activity. Labelling index was determined using radioautographs of 1 μ Epon-embedded sections. At the time of explantation the submandibular gland rudiment consisted of undifferentiated epithelial cells arranged in cords. On day 3 of culture two additional cell types could be distinguished: terminal tubule cells and proacinar cells. The proacinar cells were characterized by peroxidase activity in their granules and cytoplasm. By day 4 acinar cells begin to appear. On the fifth day of culture the four cell types of the terminal tubule were present in the following proportions: undifferentiated cells, 44%; terminal tubule cells, 19%; proacinar cells, 31%; acinar cells, 6%. These results indicate that the cytodifferentiation of the secretory unit of rat submandibular gland in vitro is comparable to the differentiation in vivo.

Notes: Adult male mice were exposed to either alternating illumination or constant illumination for 70 days. Light and dark pinealocytes were compared as to distribution within the gland and ultrastructure. Quantitative studies with the electron microscope revealed a significant reduction in pinealocyte size and Golgi complex size in constant light treatment, as well as a marked but non-significant reduction in the concentration of lipid droplets and irregular vacuoles. Under constant light treatment the cross-sectional area of pinealocyte pericapillary terminals and the number of granulated vesicles per terminal decreased significantly. A greater number of mitochondria appeared swollen, with rarified matrix and reduced numbers of cristae, with constant light treatment.These results provide ultrastructural correlation with the known reduction of pineal weight, protein synthesis and antigonadotrophic activity that is seen with constant light treatment. The marked decrease in concentration of pinealocyte granulated vesicles in constant light treatment gives morphological support to the theory that these vesicles contain antigonadotrophic secretory material.

Notes: The effects of neonatal thymectomy on the development of the lymphoid, erythroid and granulocytic cell populations in mouse bone marrow have been assessed by quantitative techniques. The numbers per unit volume of bone marrow of 17 cell types were determined in neonatally thymectomized and sham thymectomized C3H mice at two, four and eight weeks of age, and compared with those of normal C3H mice. After neonatal thymectomy the numbers of small lymphocytes, large and medium-sized lymphoid cells, and erythroid cells reached normal levels at two weeks but fell progressively to 18%, 22% and 42% of normal, respectively, by eight weeks. In sham thymectomized mice these cell populations did not differ significantly from normal. Immature and mature granulocytes were elevated in numbers two weeks after either neonatal thymectomy or sham thymectomy, suggesting a transient non-specific stimulation of granulocytopoiesis. During continuous infusion of 3H-thymidine for ten days in neonatally thymectomized mice aged four weeks and eight weeks many bone marrow small lymphocytes remained unlabeled. The results demonstrate that early postnatal development of bone marrow lymphoid and erythroid cells proceeds normally in the absence of the thymus, in accord with the concept of the bone marrow as a primary site of lymphocyte production and differentiation. In addition, some slowly-renewing small lymphocytes in bone marrow appear to be thymus-independent cells.

Notes: Short pieces of seminiferous tubules from juvenile rats were grown in tissue culture and studied by phase contrast light microscopy while living and by transmission and scanning electron microscopy after fixation and appropriate processing. The pieces of tubules remodeled in vitro, with the original explant becoming surrounded closely by a sheet of epithelioid cells and more peripherally by elongate cells. The epithelioid cells were identifiable as Sertoli cells because of the presence of characteristic Sertoli-Sertoli cell junctions near their upper surface. The elongate cells were derived from peritubular tissues, but could not be specifically identified as to cell type. Clusters of stellate cells and of round cells were present on the upper surface of the Sertoli cell sheets, but not on the elongate cells or the bare floor of the culture dish. The stellate cells were spermatogonia and the round cells were spermatocytes, as identified by fine structural features. Intercellular bridges were maintained between germ cells in culture without being surrounded by processes of Sertoli cells. Rudimentary junctions were present between germ cells and Sertoli cells in culture. The shape of germ cells in vitro was the same as the shape in situ, indicating that shape is an inherent feature of germ cells and is not determined by surrounding Sertoli cells.

Notes: The distribution of adrenergic and cholinergic nerves in the arteriovenous anastomosis in the rabbit's ear was studied light microscopically using specific histochemical techniques for catecholamines and acetylcholinesterase.Histochemical observations were made with whole stretch preparations and cryostat or paraffin sections. Both kinds of nerves showed similar distribution, though the adrenergic innervation was denser than the cholinergic one. The intermediate segments of the anastomoses, which were characterized by a very thick wall, had the most dense innervation. The small arteries and arterial segments had a moderate innervation, whereas most of the small veins and venous segments had few nerve fibers.The segmental variation in the vasomotor nerve supply clearly suggests that the intermediate segment has the most active contractility. An unusually rich innervation in the anastomosis, therefore, indicates the significance of the neurogenic mechanisms in the vasomotor control of the anastomosis.

Notes: Crystalloids of what appear to be smooth reticulum have been observed in the zona fasciculata and zona reticularis in both the stressed and nonstressed adrenal gland of the Siamese tree shrew (Tupaia glis). No crystalloids are observed in the zona glomerulosa. Similar crystalloids have been described in other steroid-secreting organs, including the antebrachial organ of the lemur (Lemur catta), the parotoid gland of Bufo alvarius and in sebaceous gland cells of the Galagos and Macaques. Moreover, the crystalloids in the present investigation resemble the paracrystalline arrays of smooth reticulum present in the adrenal cortex of the fetal guinea pig. The crystalloids show much variation in degree of organization, sometimes appearing as wavy tubules parallel with one another or as fused tubules having a “donut” configuration. In addition, the crystalloids are nearly identical to configurations which have been described in mitochondrial cristae of the protozoan, Pelomyxa carolinensis.

Notes: Mouse testes of newborn to adult were examined by freezefracture. Between the newborn Sertoli cells, gap junctions consisting of aggregations of the intramembranous particles (about 8 nm in diameter) are frequently found. Some of the junctions are about 1 μm in diameter and show particle-free regions in the aggregation. Linear arrangements of a few particles, which appear to be the initial formation of the occluding junctions, are seen in the newborn Sertoli cells. The occluding junctions are arranged in a meshwork, in which the gap junctions are situated between the stages of newborn to six days of age. The particles of the occluding junctions are predominantly located on the B face in the center of the groove instead of the A face of the ridge. The occluding junctions do not appear to surround the entire circumference of the Sertoli cell of the 6-day-old mouse. The gap junctions decrease in size. In later stages, many parallel occluding junctions (up to forty in number) are found over one Sertoli cell surface and are distributed circumferentially around the entire cell surface, indicating establishment of the blood-testis barrier. The occluding junctions dominate and the gap junctions diminish in number as development proceeds.

Notes: The application of scanning electron microscopy to the study of cell surfaces is limited in intact tissues, because extracellular material may often obscure the details of nonluminal surfaces. To remove connective tissue elements we have treated human skin and both kidney, and an autonomic ganglion of the rat with hydrochloric acid and collagenase. Regional variations in the basal surface of the nephron are noted following removal of the basement membrane. The basilar interdigitations of the cells of the proximal tubule appeared as parallel ridges encircling the tubule. Ridges on the parietal epithelium of Bowman's capsule were randomly arranged and alternated with smooth surfaces. The dermal surface of the human epidermis has an alveolar or honeycomb appearance due to the elevation of the epidermal ridges and numerous pits for the dermal pegs. At higher magnifications the basal surface of cells of the stratum germinativum possessed numerous and irregular projections. Neurons with their processes are evident in the autonomic ganglion. The soma of the neurons are enclosed by flattened satellite cells. Irregular spaces between opposed satellite cells are interpreted as regions for the passage of processes related to the ganglion cells. Nodes of Ranvier were clearly seen along nerve fibers. Some pitting of the nerve fibers was also noted. The HCl-collagenase method has the advantage of the removal of collagen and basement membrane while preserving the structural integrity of the cell surface.

Notes: In contrast to multiple injections of phenobarbital, a single injection of 50 mg/kg body wt of phenobarbital into immature male rats results, after a transient increase in hepatocyte mitotic activity, in a marked decrease in hepatocyte mitotic activity to below control levels by day 3, followed by a return to control levels by day 5. This unusual pattern of hepatocyte mitotic activity can be called forth again by a second injection of 50 mg phenobarbital/kg body wt.However, a single injection of 50 mg/kg body wt of phenobarbital into immature male rats results in a pattern of change of liver wet weight, protein, and aminopyrine demethylase activity which is similar to that observed after multiple injections of phenobarbital, except that the changes are smaller in magnitude. Liver wet weight, protein, and aminopyrine demethylase activity increase and reach a peak within two days after phenobarbital injection, and then they return to control levels by five days. The same pattern of change in liver wet weight, protein, and aminopyrine demethylase activity can be elicited again by a second injection of 50 mg phenobarbital/kg body wt.

Notes: Quantitative microscopic techniques were employed to determine relative mitotic activity of lobulo-alveolar tissue in thick sections obtained from the inguinal mammary glands of colchicine-treated post-pubertal Lewis/Mai rats sacrificed at different phases of the estrous cycle. It was found that the magnitude of lobulo-alveolar cell mitotic activity and the proportion of the lobulo-alveolar tissue with mitotic activity undergo significant fluctuations during the estrous cycle. The frequency distribution of mitotic activity in lobulo-alveolar tissue was different and heterogenous at each phase of the estrous cycle. The frequency distribution of nuclei in lobulo-alveolar tissue during the estrous cycle showed only minor variations and could not explain the variation in mitotic activity. Similarly, no morphological differences were observed in the mammary gland which could account for the variation observed in mitotic activity.It was concluded that the variation in mitotic activity of lobulo-alveolar tissue is probably related to alterations in the generation time of lobulo-alveolar cells imposed by cyclical variation in ovarian hormones during the estrous cycle.

Notes: The fine morphological and cytochemical (acid phosphatase) changes occurring during holocrine cell lysis in the sebaceous preputial gland of the adult rat were investigated. When cells reach full differentiation cisternae of the widespread rough endoplasmic reticulum show invaginations of the wall. Cytoplasmi areas comprising such altered cisternae, normal RER and some mitochondria, are later sequestered into autophagic vacuoles, either surrounded by a single membrane (type I-bodies), or by a thick wall formed by concentric membranes (type II-bodies). Other vacuoles with similarly stratified wall and containing debris and acid phosphatase appear in cells at the next stage of involution. These autophagic vacuoles (type III-bodies) are probably derived from type II-bodies, and become gradually transformed into residual bodies rich in myelin figures, while the cell membrane and the nuclear envelope rupture. These findings show that prior to cell disintegration most of the cell cytoplasm, with the exception of the lipid and proteinaceous secretion granules and some mitochondria, is focally degraded within autophagic bodies.

Notes: In placentas from viable rat fetuses which surgically had been retained beyond term in lactating animals for as long as four days, the fine structue of much of the interhemal membrane was quite like that of full-term placentas. Moreover, contrary to what has been reported previously (Jollie, 1964a), in retained placentas which were autopsied at term there were few changes in the four cytoplasmic layers of the membrane which could be interpreted as degenerative. With prolongation of the placenta beyond normal delivery the number of pericytes which were associated with the allantoic endothelium increased; the basal lamina separating the endothelium from trophoblast III progressively became more focally split; and the apposed plasma membranes between this latter layer and trophoblast II appeared to dissociate. Trophoblast II became regionally divided in its cytoplasmic organization, its more superficial zone becoming highly infolded and/or vacuolar. As a consequence, the intertrophoblastic space between layers I and II became more extensive and complex. Within this space, a flocculent material appeared; and there was ultrastructural evidence of massive pinocytosis from the space by trophoblast II. With increased prolongation of gestation, a variety of lysosome-like inclusions appeared within this layer of the membrane.

Notes: During the first four days postpartum, heterophils (polymorphonuclear leucocytes) and macrophages occur in the intercellular compartment of the luminal epithelium of the uterine endometrium. Cytochemical and ultrastructural evidence indicates that transepithelial emigration of these stromal cells to the uterine cavity is occurring. This event takes place while the luminal epithelium is proliferating in response to the estrogenic stimulus of the postpartum estrus.Heterophil emigration precedes that of the macrophages and is most conspicuous during days 1 and 2. Although it has been established that collagen fibrils occur in uterine phagocytes (Schwarz and Güldner, 1967) assumed to be macrophages (Parakkal, 1969, 1972), their precise role in collagen degradation remains undefined. It seems likely that the emigrating macrophages, heavily laden with phagolysosomal derivatives and lipid droplets, are hauling the remnants of the intercellular substance out of the endometrium during days 2-4 postpartum. Ultrastructural evidence indicates that the emigrating macrophage punctures the basal lamina and passes through the intercellular compartment of the luminal epithelium by active penetration.Another mode of macrophagic egress operates in the deep stroma of the endometrium and myometrium where lymphatic drainage occurs. Macrophages accumulate in the perilymphatic stroma as well as within lymphatic vessels.Thus macrophagic emigration through the luminal epithelium and lymphatic vessels may provide a cellular mechanism for elimination of the intercellular stromal substance in the regressing uterus. Transepithelial emigration is a mechanism which operates also in the marsupial uterus (Padykula and Taylor, 1976), and thus may be a fundamental mechanism among subprimate mammals that fulfills in part the function that menstruation effects in primates.

Notes: The principal cell types associated with the humoral immune response (monocyte-macrophages, lymphocytes, and plasma cells) are numerous in the endometrial stroma of the uterus during the first four postpartum days in two types of mammals, the marsupial North American opossum and the eutherian albino rat. This transient cellular differentiation coincides with the physiologic period of rapid uterine regression which includes massive reduction in the amount of extracellular stromal material. In addition, heterophils and eosinophils, cell types also known to be associated with phagocytic and immunologic activity, appear in the stroma during the first two postpartum days; their presence may, however, be associated more directly with the postpartum estrus that occurs on day 1 postpartum than with endometrial regression. Thus, the five cell types, which are known in pathologic conditions to be components of an inflammatory response to a foreign antigen, are conspicuously present in the normal regressing endometrium. Furthermore, there is ample ultrastructural evidence of frequent macrophagic-lymphocytic interaction, transformation of lymphocytes, and active secretion by plasma cells during this early postpartum period.An hypothesis has been derived by uniting this new description of endometrial stromal cell differentiation with the existing literature on uterine collagenase activity, an important feature of postpartum regression (reviews by Gross, 1974; Harris and Krane, 1974). It is based on the assumption that during regression the extracellular action of neutral collagenase (and possibly other extracellular proteases) release new antigenic sites in proteins located in the ground substance. In the case of collagenase, these transient antigenic sites would arise at the locus of enzymic cleavage as well as from the subsequent denaturation of the fragments of the collagen molecule. This endogenous antigenic stimulus would be strong and temporary, and would lead to the cellular manifestations of the transient humoral immunologic response which are evident in the regressing stroma of these two mammals. This humoral immune reaction may be one of the regulatory mechanisms involved in the cyclic renewal of the extracellular compartment of the uterine stroma.

Notes: The cat's tentorium cerebelli is an osseous structure which overlies much of the brain stem. None of the current brain atlases list the coordinates of this bony projection. With a stiff wire and a stereotaxic apparatus, the location of its free edge and dorsal surface was mapped in 1.0 mm steps in the skulls of 16 cats. In the midline the dorsal surface meets the calvaria at a point 7 mm posterior and 15 mm superior to the atlas zero (intra-aural plane + 10 mm horizontal). From this junction it slopes inferiorly and anteriorly between the cerebrum and cerebellum along a 50° slope to terminate along a curved line. In the midline this free edge is located 0.5 mm anterior, and 6.5 mm superior to the atlas zero. It recedes laterally along a curved line to a point 6 mm lateral, 6 mm superior and 3 mm posterior to the atlas zero. The free edge then curves forward and descends to pass through an imaginary line 3 mm anterior and 4.5 mm inferior to the atlas zero. Statistical comparisons between measurements made in male and female cats revealed no significant differences.

Notes: The effect of the monosodium glutamate (MSG) induced lesion of the arcuate nucleus on catecholamines in the arcuate nucleus and median eminence of the mouse hypothalamus was determined using the Falck-Hillarp histofluorescence technique. The number of fluorescent perikarya in the arcuate nucleus of treated animals was decreased approximately 60%; the fluorescence intensity of surviving neurons was notably reduced. These changes were accompanied by a reduction in the intensity of fluorescence in the median eminence. Pretreatment of control and MSG-lesioned animals with a monoamine oxidase inhibitor (pargyline) greatly increased fluorescence in the median eminence and arcuate nucleus of both groups. However, the number of fluorescing perikarya of the arcuate nucleus of the normal pargyline treated group far exceeded that of the pargyline MSG animals. It is concluded that neonatally administered MSG caused destruction of a large number of dopaminergic arcuate perikarya.

Notes: Chick primordial germ cells (PGCs) which separated from the “germinal crescent” entoderm in the period from stages 4 to 8 circulated mostly through the developing blood vessels from stage 10 onward and finally migrated into the gonad. The PGCs making their appearance up to this stage were generally spherical in profile, about 14 μm in diameter. Some of the PGCs, in contrast, did not enter the blood vessels but remained in the tissue (mesenchyme) of the embryo proper (tissue PGCs) and possessed pseudopodial processes, suggesting their migration by means of amoeboid movements. The circulating PGCs emerged from blood vessels in the vicinity of developing gonads by three days (gonadal PGCs). The principal mechanism responsible for the subsequent migration of gonadal PGCs is assumed to be amoeboid movements as in the case of tissue PGCs. Notable amounts of PAS-positive glycogen were demonstrated in the cytoplasm of PGCs in all stages observed. They also contained yolk and lipids intracytoplasmically, the former dissipating in relatively early stages of development. Electron microscopic observation revealed the electron-opaque, “fragmented nucleolus” in the large nucleus (8 μm in diameter), which represented another prominent feature of chick PGCs. PGCs contained a well developed Golgi complex and endoplasmic reticulum.

Notes: Biopsy and orchiectomy specimens were collected from two adult baboons (Papio anubis) at different intervals after intratesticular injection of H3-thymidine. Zenker-formol or Bouin's fixed materials were stained with PASWeigert-Hematoxylin and radioautographed using the H.S.R. (Harleco Synthetic Resin) coating technique. Morphological features of most germ cells appeared similar to those of other monkeys, except that the spermatids in steps 9 to 11 showed a spike-like projection of the acrosome. Also, the type A spermatogonia showed some resemblance to the human type A spermatogonia. The cell associations consisted of 12 stages and a large number of tubular cross sections showed the presence of two or more stages. In Papio anubis, the zygotene spermatocytes are formed in stage VIII, and spermatozoa are released during stages V and VI.H3-thymidine labeling of germ cells indicated that one cycle of spermatogenesis in this species takes approximately 11 days, and complete spermatogenesis occupies 3.8 cycles, or approximately 42 days. The presence of labeled B type spermatogonia 35 days after H3-thymidine injection indicated the existence of some stem cells. The presence of some labeled Ad spermatogonia in these specimens could not be explained. The data indicated that spermatogenesis in this monkey is somewhat different from that in other monkey species described.

Notes: Female rats were divided into six groups: (1) control, (2) one uterine artery (a.) ligated near the utero-tubal (U-T) junction, (3) one uterine a. ligated at the level of the cervix, (4) both uterine aa. ligated separately at the U-T junction, (5) both uterine aa. ligated separately at the cervix and (6) both uterine aa. tied with one ligature at the cervix. Segmental aa. were disrupted in all experimental groups except group 6. Animals were allowed to recover for ten days and killed the first metestrus thereafter. Number of eggs ovulated was determined by flushing the oviduct with saline solution and counting the ova.Control rats ovulated 5.0 ± 0.4 eggs per ovary. Groups 2 and 3 had an increase in the number of eggs shed from the ovary on the non-ligated side. In contrast, a decrease in the number of ova shed occurred on the ligated side. When both aa. were ligated separately (groups 4 and 5), irrespective of location, a decrease in the number of eggs shed by both ovaries was evident. No effect was found when only one ligature was placed near the cervix (group 6).The data demonstrate that blood supply to the ovary via the uterine artery is essential for the full complement of eggs to be shed.

Notes: In relation to chronological age and skeletal maturation, the growth of the second metacarpal is discussed based on radiographs of the right hand and wrist in 499 male and 424 female Japanese aged 1 to 18 years. When plotted against age, bone length and width present the general growth pattern of Scammon, the adolescent spurt occurring at about 13 years in the males and 11 years in the females, and the mean values are significantly larger in the males than in the females at age 15 years for length, and age 13 years for width, and later. The growth curves of both sexes are almost parallel to each other for bone length plotted against skeletal maturation. Bone width progresses in parallel in males and females until about adolescence, whereafter there is a more rapid increase in growth in the males than in the females. The mean values for length and width are almost always significantly greater in the males than in the females. The width/length index decreases rapidly until a certain period, the minimum value being at about ten years or a maturity corresponding to that age, and then increase slightly again to reach an equilibrium state on the basis of both age and skeletal maturation. At any given age, the mean values are always greater in the males than in the females. However, according to skeletal maturation, the sex differences are steadily significant at and after the skeletal maturity corresponding to about 12 years in the males and 10 years in the females.

Notes: The process of erythropoiesis and vasculogenesis in the yolk sac of the bat (Tadarida brasiliensis cynocephala) has been studied through the use of both light and electron microscopy. Stem cells arise from the leading edge of the migrating splanchnic mesoderm and transform into primitive erythroblasts. Differentiation involves either contact or association with the endodermal cells, since all erythropoietic activity occurs on the endodermal side of the expanding vascular bed, and many of the cells are in close apposition to the lateral or basal plasma membranes of the endodermal cells. Endodermal cells also phagocytize developing primitive erythroblasts during the later stage of the process when erythropoiesis is subsiding in the yolk sac.Cells destined to become the endothelium of the expanding vascular bed also arise from the leading edge of the migrating splanchnic mesoderm. Their process of differentiation involves the development of cytoplasmic extensions that may surround a group of differentiating erythroblasts, enclosing them in the newly formed lumen of the blood vessel. The cytoplasmic extensions make contact and develop junctional complexes with similar processes from other cells to complete the lumen of the lengthening vascular bed.Cells of the granulocyte series or megakaryocytes are not observed in the yolk sac of the bat as has been described in certain other species.

Notes: During the annual breeding season, the testes of the lizard Cnemidophorus gularis are yellow-orange, ovoid organs measuring almost 1 cm in greatest diameter. The pigment is confined to the testicular tunic, which contains a zone of Leydig cells and vascular channels more than 50 m̈ thick. Leydig cells constitute approximately 60% of the zone, with remaining space occupied by capillaries, sinusoids, and lymphatic vessels. Lymphatics are concentrated at the interface between tunic and seminiferous tubules. Interstitial space is poorly developed among the tubules, accounting for less than 3% of tissue volume. Capillaries, lymphatics, and a few widely scattered Leydig cells occur in the sparse interstitial space. Leydig cells in the tunic and elsewhere in the testis show ultrastructural features commonly found in mammalian Leydig cells. Separation of the tunic from the seminiferous tubules is achieved in a few seconds by manual decapsulation of the testis and yields an enriched preparation of Leydig cells that is essentially uncontaminated by tubular elements.

Notes: The morphological sequence of events occurring in the development of the lateral-line organ was investigated in the embryo of the newt, Triturus pyrrhogaster, by means of electron microscopy. Six stages of development have been defined for convenience of description. The primordium of the lateral-line organ migrating from the pre- and post-auditory placodes is segmented into a small cell mass accompanied by a few Schwann cells enveloping nerve fibers. In the clump of cells of the lateral-line primordium, three kinds of cells are already distinguished by different degrees of darkness, i.e., dark, light and slightly dark cells (stage I). Both clump and Schwann cells enter the epidermis together (stage II). The two groups of cells are separated by the inner layer of the epidermis (stage III). The organ-forming cells elongate and the inner layer of the epidermis separates the Schwann cells from the clump (stage IV). The apexes of the organ-forming cells are exposed out of the epidermis (stage V). The lateral-line organ is almost matured. The dark cells correspond to type II of the supporting cells, the light cells correspond to type I of the supporting cells and the slightly dark cells correspond to the receptor cells because of the existence of cilia and a cuticular plate. The afferent nerve endings are found on the basal surface of the receptor cell (stage VI).

Notes: Sarcoplasmic reticulum (SR) of fast extensor digitorum longus (EDL) and slow soleus (SOL) muscles of the mouse was examined by freeze-fracture techniques. A distinctive feature of sarcoplasmic reticulum from the EDL is the presence of hillocks on the A-face within the terminal cisterns. These hillocks are usually arranged in a single row which is deployed parallel to the long axis of the adjacent T-tubule. Center-to-center spacing of hillocks within a row is about 70-75 nm. Hillocks are also found scattered within the collar region. The EDL of ten week mice was characterized by sheet-like terminal and intermediate cisterns, the latter being replaced in 37 week animals by thin tubular longitudinal elements of the SR which contain no hillocks or dimples.Hillocks occur only occasionally in SR from 10 or 20 week SOL muscle. In such cases the hillocks occur singly rather than in rows as in the terminal cisterns of EDL. The predominant form of SR in the SOL contains no hillocks.Total particles in the A-face of EDL-SR (2996 particles/μ2; S.D. = ± 287) slightly exceeded that of SOL-SR (2558 particles/μ2; S.D. = ± 274 8 nm). Packing density of 8 nm particles was slightly higher for EDL (750/μ2) vs. SOL (700/μ2). The possible significance of these features of SR in fast and slow muscle is discussed.

Notes: The reproductive biology of the bat Macrotus waterhousii was investigated anatomically and physiologically several times monthly throughout the year.The male spermatogenic cycle is initiated in June; sperm are available in August. Testicular hypertrophy is accompanied by increased levels of testicular ascorbic acid and circulating plasma testosterone. Sperm are present in the seminiferous tubules and later the epididymides from August to early December. Regression in testicular size begins in late September and testes are involuted by early December. Levels of testicular ascorbic acid and plasma testosterone decline at this time, although detectable amounts of both are present year-round. Mature sperm are absent from the male tract after late November or early December.The accessory sex gland complex and Cowper's glands undergo an annual cycle in synchrony with the testicular cycle. The accessory complex contains fructose, the concentration of which varies seasonally in direct proportion to glandular hypertrophy. Only trace levels of fructose are elaborated by Cowper's glands.In contrast to vespertilionid bats, where the accessory glands remain enlarged and active when testes involute, those in Macrotus regress.The penis differs structurally from that of other temperate North American bats, primarily by lacking a bony baculum and specialized erectile accessory corpus cavernosus bodies.Males are not reproductively active in their first autumn.

Notes: The general body epidermis of the rhesus monkey (Macaca mulatta) contains no discernible melanocytes, but after repeated ultraviolet irradiation DOPA-positive melanocytes appear and increase numerically up to 30 exposures. With continued irradiation, however, the number again declines.Experiments to determine how melanogenic activity, assayed by the incorporation of labeled DOPA or tyrosine, is related to DOPA positivity indicated that biochemical activity corresponded to the histochemical pattern. Ultrastructural studies demonstrated that after the exposure to ultraviolet light, a pool of indeterminate cells in the skin of rhesus monkeys developed into melanocytes. The melanosomes formed by these cells, however, differed from the eumelanin melanosomes described in other species; they had no internal filamentous matrix with periodicity but appeared similar to phaeomelanin melanosomes. Long term ultraviolet light irradiation may damage keratinocytes and render them incapable of phagocytizing melanosomes.

Notes: Colored cold setting acrylic cement was injected through catheters into the arteries and veins of sixteen gravid uteri obtained from pigs at various stages of pregnancy. After the cement had set hard, the reproductive tracts were macerated in concentrated acid and the vascular casts resulting were washed clean of digested tissues. Small pieces of cast were coated with gold and viewed by means of a scanning electron microscope.Early in pregnancy the capillary network consists of low parallel ranks of ridges and troughs; later, cross ridging was observed which developed in complexity as pregnancy progressed. The density of capillaries comprising the microvascular network also increased towards term. No comparable changes were noted in the relatively less well vascularized areas surrounding the mouths of the uterine glands.This technique surmounted the problems of obtaining an adequate depth of focus for viewing the capillary network of the gravid uterus with the light microscope. The relationships that the blood vessels bore to the overlying epithelial layers were discussed and the inference tentatively drawn that blood flow at the capillary level was from ridge top to trough base.

Notes: Radioautography after 3H-thymidine injection, mitotic arrest by colchicine and camera lucida drawings were used to study DNA synthesis, mitosis, formation of binucleated cells and morphogenesis in the simple liver acinus of Rappaport in 10-20-day old rats. By ten days the arrangement of hepatic cell plates had already attained the adult configuration  -  irregular and thick in acinar zone 1 (periportal), straight and thin in acinar zone 3 (pericentral). The DNA synthetic index of parenchymal and bile duct cells slowly decreased during the observation period. Zonal labeling remained steady in the relationship: zone 1 〉 zone 2 〉 zone 3. Mitosis of parenchymal and bile duct cells reached a peak at 12 days, decreasing slowly thereafter. Mitosis also exhibited the relationship zone 1 〉 zone 2 〉 zone 3. The number of binucleated cells remained constant until after day 14 when it increased rapidly. Zonal distribution of binucleated cells was just the reverse of that for DNA synthesis and mitosis, that is zone 3 〉 zone 2 〉 zone 1. Radioautographic studies of binucleated cells labeled with 3H-thymidine indicated that a small percentage of them were formed by fusion of mononucleated cells.Conclusions are: (1) the 10-20-day old rat liver is expanding its cell population primarily in acinar zones 1 and 2 while overt differentiation is occurring in acinar zone 3, (2) ingestion of solid food around day 16 may be related to binucleated cell production due in part to altered portal venous blood changing the cellular microenvironments, (3) binucleated cells arise by suppression of cytokinesis, cell fusion and/or other non-mitotic routes.

Notes: The yolk sac placenta has been implicated previously in transmission of passive immunity to the fetus. This work uses an immunohistochemical technique devised by Sternberger et al. (1969) to study this problem. Rabbit serum containing gamma globulins was injected into the uterine lumen of the White Swiss mouse during the last third of pregnancy. Two or four hours later yolk sacs were removed and fixed in 2% paraformaldehyde or freeze-dried and fixed with paraformaldehyde vapors. Finely chopped tissue was treated with (1) sheep antiserum to rabbit gamma globulin, (2) an antigen-antibody complex consisting of horseradish peroxidase and rabbit anti-horseradish peroxidase and (3) hydrogen peroxide and 3,3′-diaminobenzidine. Reaction product was heavily concentrated in visceral yolk sac endodermal cells, frequently deposited in endothelial cells of vitelline vessels, and rarely in the serosal basement membrane and mesothelial cells which border the exocoelomic cavity. This supports evidence of other workers that the yolk sac membrane rapidly absorbs substances with which it comes in contact, transport into vitelline vasculature is a route of transfer from mother to fetus, and possible transfer into the exocoelomic cavity and thence to the amniotic cavity may occur in mice.

Notes: The fluorescence histochemical method of Falck-Hillarp ('62) demonstrates dense plexuses of catecholamine-containing nerves in the piloarrector muscles of adult stump-tailed macaques. When these muscle fibers are viewed under the electron microscope, most of the axon terminal profiles between them contain numerous dense-cored vesicles (adrenergic); the rest of the terminals contain mainly agranular vesicles (cholinergic). Intradermal injections of 6-hydroxydopamine (6-OHDA, 1 and 5 mg) or 5-hydroxydopamine (5-OHDA, 10 mg) into the scalp caused the fluorescence from the nerve fibers in the muscles to completely disappear. The 6-OHDA injections induced severe degenerative changes in the adrenergic terminals whereas the 5-OHDA injections caused only the granules in the vesicles to increase in numbers. Thus, the piloarrector muscles of these macaques are innervated with both adrenergic and cholinergic terminals, but the former are more numerous than the latter.

Notes: The presence of microblebs arising from the lateral surfaces and tips of microvilli of intestinal epithelial cells of thirteen-lined ground squirrels. Citellus tridecemlineatus, is reported. Microblebs measure ∼ 50 nm in diameter and are bounded by a unit membrane which is continuous with the unit plasma membrane of the microvilli. It is suggested that such microblebbing reflects the rapid turnover of microvillar contents, membrane and surface coat.

Notes: The nerves in and immediately under the epithelial layer of the trachea and primary bronchi of the cat are studied with the electron microscope. The axons are unmyelinated and contain microtubules and mitochondria. Epithelial axons are observed throughout the entire epithelial layer, but, are usually concentrated in the basal area immediately above the basal lamina. Most axons are near the basal cells; others are in the vicinity of granulated cells or occur in the intercellular spaces between other columnar cells. A few of the epithelial axons are large and crowded with axoplasmic mitochondria. The possibility that some epithelial axons may have sensory functions is discussed.

Notes: When skin from the upper lip of 12-day embryonic mice was grown for ten days in organ culture with 5.7 μg retinol added per ml of biological medium, keratinization was suppressed and a ciliated and secretory epithelium was produced. Ultrastructural features of this epithelium are described. At this very early stage mouse epidermis is thus similar to chick epidermis in its ability to undergo radical metaplasia in response to vitamin A.

Notes: This work is based on a study of 62 circles of Willis (circulus arteriosus cerebri) from patients dead from diverse causes. We have described the wide diversity of variations found to be present in the arterial vessels that form the circle, we presented the frequency of the different variations, and finally we discuss the possible ontogenic cause of these variations.

Notes: The kinetics of lymphoid cells within the epithelium of the small gut has been studied in various thymus-deprived mice and in antigen-deprived mice by the use of 3H-thymidine injections and radioautography. In thymusdeprived mice  - including adult thymectomized, thymectomized and irradiated, neonatally thymectomized, and nude mice  - and in germ-free mice decreased numbers of intraepithelial lymphocytes (IL) were found. On the other hand, the radioautographic results indicated that the remaining IL populations included both newly formed and long-lived lymphoid cells in the same percentages as found in sham-operated controls and normal mice. It is concluded that although the presence of the thymus and the antigen content of the gut is of importance to the maintenance of the numbers of cells in the lymphoid populations of the intestinal wall, the basic kinetics of these cell populations are preserved in deprived mice.

Notes: Besides the well-known striated muscle fibers, the cremaster muscle of the guinea-pig contains smooth muscle fibers, which appear either as isolated elements or as networks. These smooth muscle fibers are embedded within a matrix of the compact fibrous coat of collagenous connective tissue which covers the vaginal surface of the striated muscle layer. Presumably the smooth muscle fibers maintain the initial, fast contraction produced by the striated muscle fibers, thus attaining an economical, tonic elevation of the testicle.

Notes: The zonula occludens (Z.O.) has been implicated as a major pathway for passive fluid movement from lateral intercellular spaces to lumen in response to an expansion of blood or fluid volume. The present study examines the Z.O. of adult rat nephrons in control and Ringer-Locke or whole blood infused animals to determine whether varying the conditions of passive fluid movement would influence the structure of the Z.O. Rats were infused over 20 minutes with Ringer-Locke (7% body weight) or whole blood (2.3% body weight). In the controls, 92%-94% of all Z.O. observed in proximal tubules and 100% of Z.O. from distal tubules and collecting ducts had fused outer leaflets. The approximate linear extent of the Z.O. was 140 Å for proximal tubules, 2,500 ÅR for distal tubules and 3,000 ÅR for collecting ducts. There was no significant difference in any of these values following the infusion with either Ringer-Locke or whole blood. We conclude that expansion of blood or fluid volume causes no detectable alteration in the fine structure of Z.O. under the conditions used in the present study.

Notes: During the course of gestation, a large amount of iron is transferred rapidly and unidirectionally from mother to fetus across the placenta. It has been postulated that one of the first steps involved in placental iron transfer involves binding of the maternal transferrin-iron complex to the surface of the placenta and the subsequent removal of iron and release of transferrin back into the maternal circulation. To determine if transferrin is present on the surface of human placental villi, two different immunocytochemical methods have been used: (1) an unlabeled antibody, peroxidase-antiperoxidase (PAP) method utilizing rabbit antiserum to human transferrin, goat anti-rabbit IgG and rabbit peroxidase-antiperoxidase complex or; (2) a peroxidase-labeled antibody method utilizing goat antiserum to human transferrin and peroxidase-conjugated rabbit anti-goat IgG. The peroxidase was then localized by incubation in a diaminobenzidine-hydrogen peroxide medium. Examination of the tissue in the electron microscope revealed the reaction product deposited as discrete patches or particles on the microvillous surface of human syncytial trophoblast. Controls using non-immune serum or an antiserum adsorbed with purified human transferrin showed no reaction product on the surface. The results provide morphological confirmation for the presence of transferrin on the surface of human syncytial trophoblast lining the maternal blood spaces.

Notes: Monosodium glutamate was administered daily on days 5 through 10 postnatally at a dose of 2.5 mg/gm body weight. Counts of remaining perikarya in the arcuate nucleus of adult mice indicated approximately an 80% decrease in the number of perikarya. The arcuate lesion resulted in endocrine deficits; reproductive capacity was reduced, animals were smaller in stature and obese, and the weights of the anterior pituitary, ovaries and testes were significantly decreased while adrenals were unaffected. Light microscopic studies revealed no significant changes in thickness or general histological appearance of the median eminence. At the electron microscope level, there were no alterations in the number of nerve terminals or dense core vesicles per unit area in the contact zone. These observations suggest that afferents to the median eminence from the arcuate nucleus may form a relatively small portion of its total nerve terminal population.