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The initial segment of the epididymis of rats, fixed with glutaraldehyde, was postfixed with reduced osmium, a technique that clearly delineates the membranes of cisternae of the endoplasmic reticulum (ER) and the various elements of the Golgi apparatus, or with tannic acid to enhance the coats of vesicles and ribosomes on ER cisternae. The material was also treated to demonstrate various phosphatase activities (NADPase, TPPase, CMPase, G-6-Pase) or impregnated with osmium tetroxide. In osmium-impregnated material, the Golgi apparatus of the epithelial principal cells of the initial segment appeared in the light microscope as a branching, anastomosing ribbon forming a large network in the supranuclear region. In the electron microscope, ER were of two types: the heavily granulated, flattened, rough ER seen in the infranuclear and juxtanuclear regions and the distended, tubular, sparsely granulated ER, showing only few ribosomes, seen interlaced with the Golgi ribbon in the supranuclear region and at the apical pole of the cell. Of particular interest in this cell was the fact that the sparsely granulated ER approximated the Golgi stack on both its cis- and trans-faces. On the cis-face of the Golgi stack, the sparsely granulated ER cisternae showed the usual finger- or bud-like protrusions directed toward the cis element of the Golgi stack and around which numerous small 80 nm vesicles or membranous tubules were clustered. The Golgi stack consisted of the following elements in a cis-trans axis: the cis osmiophilic element, a first saccule slightly dilated, saccules two to four (S2-S4), which were NADPase-positive, and saccules five to seven and the eight Golgi element, which were TPPase-positive. On the trans-aspect of the Golgi stacks, several (up to four) CMPase-positive trans-Golgi networks were observed often in close apposition to the sparsely granulated ER cisternae. One of the trans-Golgi networks showed a "peeling-off" configuration, i.e. part of it was closely apposed to the overlying Golgi element of the stack, whereas the remaining part was separated from the stack by a space occupied by a cisterna of sparsely granulated ER. The other trans-Golgi networks were completely separated from the stack and were often seen sandwiched between sparsely granulated ER cisternae. Thus, ER cisternae showed extensive areas of close apposition but no continuity with the trans-Golgi networks. Although the saccules of the Golgi stacks showed NADPase and/or TPPase activity, the trans-Golgi networks displayed CMPase activity, thus facilitating their identification from the closely associated unreactive sparsely granulated ER cisternae.(ABSTRACT TRUNCATED AT 400 WORDS)
Anat Rec 1991 Feb
PMID:Golgi apparatus of epithelial principal cells of the epididymal initial segment of the rat: structure, relationship with endoplasmic reticulum, and role in the formation of secretory vesicles. 184 81

Ten male goats and five rams were examined from 11 and 15 weeks of age, respectively, for six months to study the ultrasonic appearance of normal testes and epididymides before and after puberty. Five adult rams with lesions of these organs were also examined. A portable, B-mode, real time scanner fitted with a 7.5 MHz, linear array transducer was used. The testis appeared as a homogeneous and moderately echogenic structure with a centrally located mediastinum testis represented by an hyperechogenic line in images taken in the longitudinal plane and by an almost circular spot in transverse images. The testicular capsule and skin were evident as a distinct hyperechogenic line encircling the testicular parenchyma. A thin non-echogenic layer of fluid, presumably between two layers of tunica vaginalis, was observed. The tail of the epididymis was more heterogeneous and less echogenic than the testis. The epididymal head was also less echogenic but homogeneous in texture, and the body of the epididymis was difficult to image. The pampiniform plexus was easily identified as numerous convoluted sonolucent tubular structures. The ultrasonic images of possible cases of epididymitis, spermatocele, testicular cyst and abscess and scrotal hernia are described.
Vet Rec 1991 May 25
PMID:B-mode real time ultrasonographic imaging of the testis and epididymis of sheep and goats. 186 75

The distribution of actin in spermatogenic cells and epididymal spermatozoa of the opossum, Monodelphis domestica, was examined by immunofluorescence microscopy to identify its potential function in the major structural events of sperm development. In spermatogenic cells actin was located at the site of initial interaction between the nucleus and acrosome and remained present through subsequent acrosome morphogenesis. Actin was also associated both with the posterior pole of the nucleus, at the site of flagellar attachment, and with the manchette. Thus actin may play a role in establishing the specific associations of spermatid organelles and in the streamlining of the cells' architecture. In epididymal spermatozoa two sites of actin localization are present. The first site is surrounding the connecting piece where it may participate in the characteristic 90 degrees rotation of the head. The second site was a ring of actin surrounding the lateral boundary of the acrosome where it may play a role in the sperm pairing process which also occurs in the epididymis.
Anat Rec 1991 Jun
PMID:Changes in actin distribution during sperm development in the opossum, Monodelphis domestica. 186 97

The scanning and transmission electron microscopes were used to examine the processes of spermiation and sperm maturation in the marmoset. We observe that the heads of late spermatids are embedded in the apical aspect of the large sleeve-like columnar portion of Sertoli cells. As spermiogenesis progresses, spermatids become associated with numerous small apical Sertoli cell extensions. These finger-like processes undergo a sequence of changes during spermiation. Spermatozoa from the caput, corpus, and cauda epididymides were examined. In caput epididymis of marmoset, the apical segment of the spermatozoa extends well beyond the rostral edge of the nucleus and folds back on itself. In sagittal sections, the acrosome exhibits a distinct hook shape. In the corpus, the distinctive hook-shaped apical segment of the acrosome is observed in some spermatozoa, but the apical extension is significantly smaller or projects out only slightly beyond the nucleus. In cauda epididymis, the extension is absent. A similar acrosomal hook has been reported in the pigtailed monkey, which is an Old World species. We suggest that changes in acrosome structure during sperm maturation may be fairly widespread among primates.
Anat Rec 1991 Mar
PMID:Spermiation and sperm maturation in the marmoset. 190 30

Using specific polyclonal antibodies generated against a 13 KD human testicular inhibin, immunocytochemical localization of inhibin was carried out in different regions of human epididymis. The concentrations of inhibin were greater in caput and corpus regions as compared to the caudal region. The epididymal inhibin was found to be bioactive, since it suppressed specifically the FSH levels of rat pituitaries in vitro. Spermiophage/macrophage cells exhibited strong staining for inhibin which were suggestive of a possible role of inhibin in modulation of immune function. In view of the known activities of inhibin in cellular growth, differentiation, and steroidogenesis, epididymal inhibin could have a role in acquisition of sperm fertilizing capabilities.
Anat Rec 1991 Aug
PMID:Occurrence of bioactive and immunoreactive inhibin (13 KD) in human epididymis. 192 51

During epididymal transit, the mouse sperm flagellum acquires a surface glycoprotein (SMA4) from epididymal fluid that functions as a sperm antiagglutinin. To determine the origin of this molecule, testes and epididymides of male mice were sectioned for light microscopy and stained with wheat germ agglutinin (WGA)-peroxidase, a probe that has been used previously to examine the biology of SMA4. WGA reactivity was localized to the cytoplasm in a small population of cells in the distal caput epididymis. Testis cells and principle cells of the caput were nonreactive with WGA, while stereocilia were stained on principle cells in the corpus and cauda. The WGA-positive cells in the distal caput were identified as holocrine cells on the basis of morphology, distribution, and PAS + reaction. At high magnification, intense WGA reactivity was due to the presence of numerous apical granules in the cytoplasm. The location of the cells in distal caput coincided exactly with the region of tubule in which sperm first acquired SMA4 on their flagellae. These data suggest that holocrine cells near the junction of caput and corpus epididymis are the source of the sperm antiagglutinin SMA4.
Anat Rec 1987 Feb
PMID:Maturation antigen of the mouse sperm flagellum: II. Origin from holocrine cells of the distal caput epididymis. 303 4

Rat spermatozoa from the epididymis and ductus deferens were observed by surface replica, rapid-freeze and deep-etch, and conventional freeze-fracture methods. By the surface replica method, parallel periodical ridges were observed in the acrosomal region of the spermatozoa from the distal part of the cauda epididymis (zone 6) and from the ductus deferens. The periodicity of the ridges forming a domain was about 35 nm. A quantitative analysis of the spermatozoa along the reproductive tract indicated that 39.4% and 73.5% of the population in zone 6 of the epididymis and in the ductus deferens, respectively, had the domain. None of the spermatozoa from zone 1 through zone 5 had the domain. The results of the rapid-freeze and deep-etch procedure showed that the ridges observed by the surface replica method consisted of linear arrangements of elliptical particles on the ES face of the plasma membrane. The particles were about 30 nm in length and 15 nm in width. On the corresponding PF face of the plasma membrane, linear arrangements of the intramembrane particles (IMPs) of about 8 nm in diameter were observed by both the deep-etch and freeze-fracture methods. The IMPs tended to run in paired parallel lines. A close relationship was observed between the lines of the elliptical particles on the ES and of the IMPs on the PF faces. The elliptical particle may be a protruded part of the IMP(s) or other protein(s) bound to the IMP(s).
Anat Rec 1988 Jan
PMID:Maturation changes of the plasma membrane of rat spermatozoa observed by surface replica, rapid-freeze and deep-etch, and freeze-fracture methods. 334 86

Testicular feminization (Tfm) in the mouse is characterized by androgen insensitivity of the target cells. We describe the presence of androgen-insensitive Tfm cells in the epididymis of mosaic mice produced by converting female carriers of the Tfm mutation (XTfm/X+) to males via the sex reversal factor (Sxr). The mosaic arises by random X-inactivation. In the epididymal duct, flat undifferentiated Tfm cells are interspersed between high columnar wild-type cells. By thaw-mount autoradiography we show that after injection of [3H]dihydrotestosterone, radioactivity is concentrated in the nuclei of high columnar wild-type cells, whereas the nuclei of the low cuboid Tfm cells remain free of nuclear radioactivity. After injection of [3H]estradiol, both Tfm and wild-type cells show nuclear labeling. Our observations demonstrate that Tfm cells in the mosaic epididymis selectively lack nuclear dihydrotestosterone-binding sites, whereas estradiol-binding sites are intact.
Anat Rec 1988 Apr
PMID:Androgen receptor-deficient Tfm cells in the mosaic epididymis of sex-reversed mice heterozygous for Tfm: an autoradiographic study with [3H]-dihydrotestosterone and [3H]-estradiol. 338 28

Purified boar sperm plasma membranes (PM) and PM proteins were used as antigens to produce 58 monoclonal antibodies against surface antigens. Fluorescence labelling (biotin-avidin-FITC) was used to determine the distribution of antigens in caput and cauda epididymal and in ejaculated spermatozoa with hybridoma supernatants and/or 1:100 diluted ascites fluid after subcloning. Sixteen areas (subdomains) of apparent restricted antigen mobility were identified and significant differences in the localization of most antigens in caput, cauda, and ejaculated PM were recognized. While localization patterns were highly reproducible with a given protocol for sample preparation and immunolabelling, localization patterns were markedly affected by changes in protocols. Fluorescence patterns were affected by the manner in which sperm were labelled (live sperm or sperm labelled at various steps), by washing, and by temperature or by addition of seminal plasma. These results indicate that the dynamic properties of the sperm PM or the surrounding fluids can easily mask or unmask or reconfigure binding sites for highly site-specific monoclonal antibodies and that antigen distribution is probably under-estimated when these labelling techniques are used. Such changes in the accessibility of antigenic sites to monoclonal antibodies limited determining the extent of distribution of a given antigen on epididymal sperm. However, the reproducibility of patterns when a given protocol is used and the large number of antibodies (39/42) displaying marked differences in localization on caput, cauda, and ejaculated PM suggest that changes in the organization of the PM constituents, whether by addition or subtraction of antigen or through configurational changes in proteins, are a major consequence of sperm maturation in the epididymis.
Anat Rec 1986 Mar
PMID:Immunofluorescence antigen localization on boar sperm plasma membranes: monoclonal antibodies reveal apparent new domains and apparent redistribution of surface antigens during sperm maturation and at ejaculation. 351 13

Vasoactive intestinal polypeptide (VIP) and enkephalins were demonstrated in the nerves of the human male urogenital tract by light and electron microscope immunohistochemical techniques. Nerves containing immunoreactivity to VIP were more numerous than enkephalin-immunoreactive nerves. Both VIP- and enkephalin-immunoreactive nerves were detected in the vas deferens, prostate, seminal vesicles, and urinary bladder. In the kidney, testis, and epididymis no immunoreactive nerves could be demonstrated. By electron microscope both types of immunoreactivities were localized to the large granular vesicles of nerve terminals. VIP-immunoreactive nerves were mostly found subepithelially, whereas enkephalin-immunoreactive nerves were mainly related to smooth muscle cells. The possible functions of these peptide-containing nerves are discussed.
Anat Rec 1986 May
PMID:Light and electron microscope demonstration of VIP- and enkephalin-immunoreactive nerves in the human male genitourinary tract. 351 43


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