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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 109 (1998), S. 431-447 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The Golgi apparatus of epididymal principal cells shares many structural features with other cell types. Saccular regions are arranged in a cis-Golgi network, eight flattened saccules, and several trans-Golgi networks (TGNs). Dilated tubules form intersaccular connecting regions which joint together saccules at the same or different levels between adjacent stacks. Wells exist as large perforations in register with the four cis-most saccules and serve as areas of vesicular interactions. TGNs are variable and can appear to peel off the stack or to be detached from it in the form of an anastomotic tubular network with pale dilated areas corresponding to prosecretory granules connected by short narrow bridges. Elongated or discoid dilated cisternae of endoplasmic reticulum (ER) (sparsely granulated) lie over the cis face of the stack, from which they are separated by an intermediate compartment filled with vesicles and tubules. The ER is also closely juxtaposed to the TGNs and the eighth saccule but interconnections are never seen between them. Vesicles of the COP variety reside at all levels of the stack and appear to bud off the cis-located ER and the edges of the saccules, while clathrin-coated vesicles appear mainly on the trans face of the stack and next to lysosomes. In the supranuclear cytoplasm, clusters of vesicles and tubules, at times budding off enveloping ER, appear to radiate toward the Golgi stacks where they fuse with cis Golgi elements. Taken together, these observations suggest dynamic functions and interactions for the various Golgi elements, associated vesicles, ER, and vesicular tubular clusters.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 40 (1995), S. 69-83 
    ISSN: 1040-452X
    Keywords: SGP-1 ; Hypophysectomy ; Castration ; Efferent ducts ; Lysosomes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The objective of this study was to define the factors regulating the endogenous production of sulfated glycoprotein-1 (SGP-1) in nonciliated cells of the efferent ducts. To this end we examined five different groups of animals undergoing the following experimental procedures: (1) hypophysectomized animals at 7, 14, and 28 days, (2) 7-day hypophysectomized rats receiving testosterone implants given at various time intervals thereafter, (3) castration at various time intervals up to 7 days, (4) 7-day castrated rats receiving testosterone implants at various time intervals thereafter, and (5) castrated rats given testosterone implants immediately after castration and sacrificed at different time intervals thereafter. Efferent ducts were fixed by perfusion with 4% paraformaldehyde and 0.5% glutaraldehyde in phosphate buffer for quantitative immunocytochemical analysis at the level of the electron microscope. For each experimental condition and their controls, the number of gold particles/μm2 within the endosomal and lysosomal compartments was calculated taking into account the changes in both the volume of the cell and organelles being quantified and expressed as labeling content. The results revealed that hypophysectomy (up to 4 weeks) caused a marked significant decrease in the SGP-1 labeling content of the endosomal and lysosomal compartments. The labeling content of the lysosomal compartment of efferent ducts from rats castrated for up to 1 week did not change significantly. However, there was a significant decrease in the labeling content of endosomes. This decrease is due to SGP-1, which is secreted by Sertoli cells, not being available for uptake in the efferent aucts. These results suggested that testosterone is not required for maintaining the high labeling content of SGP-1 within lysosomes of nonciliated cells, but that a pituitary factor appears to be needed. The administration of testosterone at different intervals to 7-day castrated animals resulted in a significant decrease of lysosomal SGP-1, suggesting that testosterone under these experimental conditions inhibits the production of a pituitary factor that maintains the high labeling content of SGP-1 within lysosomes of the nonciliated cells. Testosterone administered to 7-day hypophysectomized animals over a 24-hr period had no effect on the labeling content of SGP-1 within lysosomes. However, the administration of testosterone to animals immediately following castration showed no differences in the labeling content of SGP-1 within compared to controls. Together these results suggest that the labeling content of SGP-1 within lysosomes of nonciliated cells of the efferent ducts is not dependent on luminal or circulating androgens, nor is it dependent on a testicular factor entering the lumen of the ducts. It does appear, however, that SGP-1 synthesis and targeting to secondary lysosomes is dependent on a pituitary factor that may have a direct or an indirect effect on the nonciliated cells. © 1995 Wiley-Liss, Inc.
    Additional Material: 13 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 41 (1995), S. 287-299 
    ISSN: 1040-452X
    Keywords: SGP-1 ; SGP-2 ; Postnatal development ; Nonciliated cells ; Efferent ducts ; Rats ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sulfated glycoprotein (SGP) -1 and -2, secretory products of Sertoli cells, are secreted into the lumen of seminiferous tubules where they bind to late spermatids. Once released, the spermatozoa traverse the efferent ducts where these proteins detach from their surface and are endocytosed by the nonciliated cells. In adult animals, SGP-1 and SGP-2 are also synthesized by nonciliated cells and targeted from the Golgi apparatus to lysosomes. The purpose of the present study was to determine the pattern of expression of SGP-1 and SGP-2 within nonciliated cells during postnatal development. The efferent ducts of animals at different postnatal ages were prepared for an electron microscopic immunocytochemical quantitative analysis as well as for Northern blot analysis. The data expressed as labeling content (no. gold particles/μm2 and taking into account the volume of the endocytic or-ganelles and the cell) revealed that anti-SGP-1 labeling in endosomes of nonciliated cells was minimal at 15, 21, and 29 days of age. On the other hand, the lysosomal labeling content showed a significant increase by day 29 compared to 15 and 21-day-old animals indicating that an endogenous form of SGP-1 was being synthesized by nonciliated cells and targeted to lysosomes. By day 39 a significant increase in endosomal labeling occurred; this was attributed to the endocytosis of Sertoli-derived SGP-1 which coincided with the entry of spermatozoa into the lumen of these ducts at this age. Lysosomal labeling showed further significant increases at days 39, 49, and then again at day 90. Northern blot analysis detected SGP-1 mRNA transcripts at all postnatal ages examined. While decreases or increases in transcripts could not be determined due to the greater amount of tissue present with increasing age, these data taken together support the idea of an endogenous form of SGP-1 being synthesized by nonciliated cells and targeted to lysosomes during postnatal development.In the case of SGP-2, endosomal labeling was minimal at 15, 21, and 29 days of age but was significantly increased by day 39, with similar values at all subsequent ages. The high value at day 39 was attributed to the endocytosis of SGP-2 which coincided with the entry of spermatozoa into the lumen at this age. Lysosomal labeling, on the other hand, was low at days 15 and 21 but peaked at day 29 at a time when endosomal labeling was minimal. These results suggested the synthesis of an endogenous form of SGP-2 which was being targeted to lysosomes. Similar values for SGP-2 lysosomal labeling comparable to that at day 29 were obtained at all other ages. Since SGP-2 endosomal labeling was significantly increased at day 39 and maintained thereafter, it is suggested that labeling in lysosomes at this and subsequent ages could also be due to the endocytosis of Sertoli-derived SGP-2. However, Northern blot analysis confirmed the presence of mRNA transcripts for SGP-2 at all postnatal ages examined, although increases or decreases in their amount were not determined. These results thus consolidate the hypothesis of an endogenous form of SGP-2 being synthesized by nonciliated cells and targeted to lysosomes. Finally, since the amounts of endogenous SGP-1 and SGP-2 peak at different ages, it is suggested that different factors are involved in regulation of these two proteins during postnatal development. © 1995 Wiley-Liss, Inc.
    Additional Material: 12 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 242 (1995), S. 289-301 
    ISSN: 0003-276X
    Keywords: Golgi apparatus ; Trans-Golgi networks ; Secretory granules ; Three-dimensional electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The trans-Golgi network (TGN) is generally considered as a distinct and permanent structural compartment of the Golgi apparatus of various cell types. To verify this postulate we examined and compared the three-dimensional characteristics of the TGNs of 14 different mammalian cell types as presented in our various publications since 1979 when we initially described the trans-tubular network of Sertoli cells.Methods: In all these studies we used low and high voltage electron microscopes on thin or thick sections of tissues fixed with glutaraldehyde and postfixed with reduced osmium. The section were stained with uranyl acetate and lead citrate. Stereopairs, prepared from photographs of tilted specimens, permitted a direct observation of the three-dimensional structure of the various elements of the Golgi apparatus.Results: The TGNs are multilayered and extensive in cells which do not form large typical secretory granules (Sertoli cells, nonciliated cells of ductuli efferentes, spinal ganglion cells) but have an extensive lysosomal system. The TGN is absent in cells forming very large secretory granules (secretory cells of seminal vesicles and lactating mammary glands). The TGNs are small in cells producing small to medium-size secretory granules and/or appear as residual fragments on the trans aspect of the Golgi stacks (e.g., mucous cells of Brunner's gland, pancreatic acinar cells, etc.). In cells with multiple and extensive TGNs, a continuity of these tubular networks with the two or three transmost saccules of the stack is observed but there are seemingly no connections between the TGNs. Whenever the TGNs are present, they do not form a continuous structure along the Golgi ribbon. However, they do present, in all cases, configurations suggestive of desquamation and renewal.Conclusions: The structure of the TGN varies considerably from one cell type to another, being extensive in cells not showing typical secretory granules but having an extensive lysosomal system, while in secretory cells showing small or large secretory granules the TGN is either small or even entirely absent. © 1995 Wiley-Liss, Inc.
    Additional Material: 14 Ill.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 242 (1995), S. 515-530 
    ISSN: 0003-276X
    Keywords: Intermediate zone ; Epididymis ; Principal cells ; Giant endosomes ; SGP-2 ; Immobilin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: In the present study, principal cells of the intermediate zone of the epididymis, an area situated between the initial segment and proximal caput, were observed to present morphological features distinct from those of principal cells of other regions.Methods: The epididymides of adult rats were fixed by perfusion with glutaraldehyde and embedded in Epon. Administration of fluid phase tracers was performed in the case of several animals. Localization of anti-SGP-2 and anti-immobilin antibodies in conjunction with light (LM) and electron (EM) microscope immunocytochemistry was also performed.Results: In the LM and EM, the most distinctive feature of many principal cells of this zone was the presence of apically located vacuoles referred to as giant endosomes due to their large size and because they readily incorporated tracers introduced into the lumen of the epididymal duct and were acid phosphatase-negative, Giant endosomes, containing electron-dense granular patches, appeared to form by the progressive fusion of small, medium, and large endosomes. In the supranuclear region, multivesicular bodies (MVBs) and lysosomes were present. Although smaller in size than the giant endosomes, MVBs and lysosomes contained the electron-dense patches. It is suggested from morphological images that giant endosomes fragment into smaller units corresponding to MVBs which gradually transform into lysosomes. Experiments using anti-SGP-2 and anti-immobilin antibodies revealed gold particles over the Golgi apparatus and secretory vesicles (150-300 nm) of principal cells of this zone as well as the luminal contents indicative of secretion of these proteins. Interestingly, giant endosomes were also immunolabeled with both antibodies as were stereocilia, coated pits and vesicles, and endosomes of various sizes; lysosomes were minimally labeled. These results suggest that principal cells of the intermediate zone endocytose as well as secrete SGP-2 and immobilin. The internalized SGP-2 and immobilin may correspond to that secreted further upstream and that, possibly due to their short half-life and terminated function, are removed from the lumen of the duct. Principal cells of this zone secrete these proteins possibly to replenish that lost by endocytosis.Conclusions: Principal cells of the intermediate zone contain giant endosomes. The presence of such large structures suggests that the early events in endocytosis is a slower process in principal cells of this zone as compared to other regions. The fact that these cells both secrete and endocytose SGP-2 and immobilin adds to the complexity of our understanding of how principal cells function along the length of the epididymis. © 1995 Wiley-Liss, Inc.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 209 (1984), S. 185-195 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The endocytic activity of the low cuboidal cells lining the rete testis was analyzed by electron microscopy following injection of various tracers into the lumen of these anastomotic channels. At 1 and 5 minutes after injection, cationic ferritin (CF) and concanavalin A-ferritin (Con A) were seen bound to the apical plasma membrane and to the membrane of subjacent vesicles or invaginations connected to this apical membrane. At 30 and 60 minutes, these tracers were found in intracytoplasmic vesicles and in vesicles connected to the lateral or basal plasma membrane as well as in the lateral intercellular space and in the lamina lucida of basal lamina. At 30 minutes, CF and Con A also appeared in the matrix of pale multivesicular bodies while at 1 hour dense multivesicular bodies were labeled. At 2 hours and later time intervals, the tracers accumulated in dense granules identified as lysosomes. Native ferritin (NF), concanavalin A-ferritin in presence of α-methyl-D-mannoside, and horseradish peroxidase or albumin bound to colloidal gold were all to be incorporated by the lysosomal system of these epithelial cells, as just described for CF and Con A, but these various tracers were not bound to the apical plasma membrane or to the membrane of cytoplasmic vesicles, nor were they found in the intercellular spaces or the lamina lucida at the base of the cells. Thus, the epithelial cells of the rete testis do not appear to be only involved in the uptake of substances from the lumen and their disposal by the lysosomal system, but also appear to contribute to the transport of certain macromolecules from the lumen to the laterobasal surfaces of the cells. These cells may thus play a role in determining the composition of the rete testis fluid.
    Additional Material: 15 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 196 (1980), S. 83-99 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Throughout stage VII and early stage VIII of the cycle of the seminiferous epithelium, the heads of the late spermatids, located in a juxtaluminal position, are embedded in apical processes of Sertoli cells. These processes contain cisternae of endoplasmic reticulum (ER) of two main types, i.e., flattened and tubular, which communicate with each other to form a continuous system. Throughout the long stage VII of the cycle, these two types of cisternae undergo marked changes. In early stage VII, the flattened cisternae, developing from the subsurface cisternae which compose the “junctional specialization,” form concentric sheets at the periphery and in the middle of each apical process. The less conspicuous tubular cisternae form a continuous network which is present in the bridge connecting the Sertoli cell body to the apical process, and extends along the dorsal and ventral aspects of the spermatid's head to end up as cup-shaped flattened cisternae capping the bulbs of the tubulobulbar complexes described by Russell and Clermont ('76). In mid stage VII, the flattened cisternae start to regress, while the tubular cisternae become more abundant. In late stage VII, only fragments of the flattened cisternae are present, while the tubular cisternae form a profuse and elaborate network throughout the apical process. In the following stage VIII, the tubular cisternae disperse and only remnants of ER are present at the time of the release of the spermatid into the tubular lumen. These transformations of ER cisternae suggest a complex alteration in the relationship between Sertoli cells and late spermatids prior to their release as spermatozoa.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 157 (1980), S. 357-373 
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Glutaraldehyde-fixed testes were impregnated with the Ur-Pb-Cu technique of Thiéry and Rambourg (′76) or postfixed in ferrocyanide-reduced osmium (Karnovsky, ′71). Thin and thick (0.5 μm) sections were examined with a Philips 400 electron microscope at 80 or 100 kv. Stereopairs were prepared from pictures of the same field after tilting the specimen every 6° from the -45 degree to the +45 degree position of EM goniometric stage. The cortex of the compact hemispherical Golgi apparatus of young spermatids (steps 2-8) was found to be composed of saccular and intersaccular regions similar to those described in the Golgi apparatus of Sertoli cells (Rambourg et al., 1979). In the saccular region, the stacks were composed of three to nine parallel saccules perforated with pores of various dimensions. On the mature or trans-face of the stack, one or two membranous elements with a wider lumen were either closely applied to the overlying saccules or were separated from them and intermixed with the vesicular components of the medulla. On the forming or cis-face of the stack, three or four saccules were frequently interrupted by gaps in register from one saccule to another. In three dimensions, these gaps appeared as pan-shaped spaces or “wells,” often containing a few vesicles. Immediately overlying the first saccule on the cis-face, a regular network of anastomotic tubules was present, corresponding to the cis-osmiophilic element observed in other cell types. In the intersaccular region, membranous tubules connected to the edges of the saccules branched, intertwined, anastomosed, and bridged adjacent stacks of saccules. Such membranous tubules bridged saccules with the cis-osmiophilic element or saccules of the same stack. Between the ER cisternae capping the surface of the Golgi apparatus and the cis-network of anastomotic tubules, there was a space called the peripheral Golgi region containing small vesicles and membranous tortuous tubules. The vesicles were frequently arranged in clusters that were capped by an ER cisterna and displayed a size gradient from the periphery to the center of the cluster. Thus, although there were similarities between the three-dimensional architectures of the Golgi apparatus in Sertoli cells and young spermatids (e.g., saccular and intersaccular regions), several structural features distinguished the spermatid's Golgi apparatus.
    Additional Material: 15 Ill.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 171 (1984), S. 41-57 
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The “round body,” a spherical structure typically associated with a nucleolus in male germ cells of the rat, has been examined in the electron microscope using routine and cytochemical methods to determine its structure, composition, and mode of development. Cytochemical analysis indicates that the round body includes neither nucleic acid nor lipid, but is composed of nonhistone protein which appears in the form of 1.6-nm-wide fibrils. Development begins in late leptotene, when a single round body appears in each spermatocyte as an irregular spheroid located along the inner surface of the nuclear envelope. During subsequent stages of the meiotic prophase, the round body leaves the nuclear envelope, becomes a regular sphere, and gradually enlarges from a diameter of 0.4 μm in leptotene to 1.6 μm in diplotene. Concurrently, lacunae appear within its substance and enlarge. At each maturation division, the amount of round-body material is decreased by about half, presumably because the constituent proteins are dissociated at metaphase, distributed between the two daughter cells at telophase, and reconstituted into half-sized round bodies. As spermiogenesis proceeds, the round body shrinks gradually and disappears at step 8.Soon after its appearance at leptotene, the round body becomes associated with and is surrounded by the pars granulosa of one of the nucleoli. Moreover, 3H-uridine incorporation into nucleolar RNA is high as long as the size of the round body increases, but is low or absent when it decreases. It is possible, therefore, that the round body exerts some control on nucleolar activity in meiotic cells.
    Additional Material: 26 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 171 (1984), S. 59-74 
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The nonciliated cells lining the ductuli efferentes presented three distinct cytoplasmic regions. The apical region contained, in addition to cisternae of endoplasmic reticulum and mitochondria, two distinct membranous elements. The tubulovesicular system consisted of dilated tubules connected to the apical plasma membrane and subjacent distended vesicular profiles. The apical tubules, not connected to the cell surface, consisted of numerous densely stained tubules of small size which contain a compact, finely granulated material. The supranuclear region, in addition to a Golgi apparatus and ER cisternae, contained dilated vacuoles, pale and dense multivesicular bodies, as well as numerous dense granules identified cytochemically as lysosomes. The basal region contained the nucleus and many lipid droplets. The endocytic activity of these cells was investigated using cationic ferritin (CF) and concanavalin-A-ferritin (Con-A-ferritin) as markers of adsorptive endocytosis; and native ferritin (NF), concanavalin-A-ferritin in the presence of α-methyl mannoside, and horseradish peroxidase or albumin bound to colloidal gold for demonstrating fluid-phase endocytosis. These tracers were injected separately into the rete testis, and animals were sacrificed at various time intervals after injection. At 1 min, CF or Con-A-ferritin were seen bound to the apical plasma membrane, to the membrane of microvilli, and to the membrane delimiting elements of the tubulovesicular system. Between 2 and 5 min, these tracers accumulated in the densely stained apical tubules and at 15 in min the dilated vacuoles. Between 30 min and 1 hr, the tracers appeared in multivesicular bodies of progressively increasing density, whereas at 2 hr and later time intervals, many dense lysosomal elements became labeled. The tracers for fluid-phase endocytosis showed a distribution similar to that for CF or Con-A-ferritin except that they did not bind to the apical plasma membrane, microvilli, or membrane delimiting the tubulovesicular system. At no time interval were any of the tracers observed in the abluminal spaces. Thus, the nonciliated epithelial cells of the ductuli efferentes are actively involved in fluid-phase and adsorptive endocytosis, both of which result in the sequestration of endocytosed material within the lysosomal apparatus of the cell.
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