Lysosomal integral membrane proteins exhibit region and cell type specific distribution in the epididymis of the adult rat.

The epididymis, a post-testicular site required for maturation and storage of spermatozoa, is actively involved in exocytic and endocytic events, two phenomena likely to depend on the integrity of the lysosomal system. To study the lysosomal system of the epididymis, five monoclonal antibodies, previously characterized as recognizing five distinct lysosomal integral membrane proteins (LIMPs 1-5), were used as molecular probes of lysosome distribution in cells lining the epithelium. Immunocytochemical localization of LIMPs, using biotin-streptavidin immunoperoxidase methodology, was performed on frozen sections of adult rat epididymides and in cell cultures prepared from either the caput or cauda epididymis. In frozen sections, a heterogeneous distribution of the different LIMPs along the length of the epididymis was observed. For example, the distribution of LIMP 1 (35-50 K) was detected in all cells of the caput and quite dramatically in clear cells of the distal caput, corpus, and cauda epididymis, but specifically not in the principal cells of the distal caput, corpus, and cauda. In contrast, LIMP 2 (64-71 K) was present in all cells of the epididymis, except clear cells. LIMPs 4 and 5 (93 K and 93 K) were detected in all epididymal cells, including the clear cells. Finally, whereas the regional and cell type distribution of LIMP 3 (74 K) in the epididymis was identical to that of LIMPs 4 and 5, the nature of the vesicles immunostained was distinct. In cultured cells, the general immunostaining patterns observed in vivo were maintained during the duration of the primary cultures for all five LIMPs. Our results begin to address the molecular heterogeneity of the lysosomal system along the length of the epididymis, and may suggest in part a basis for underlying structural and functional characteristics of the epididymis leading to the sequential maturation of sperm.     

Histochemical evaluation of glycoconjugates in the male reproductive tract with lectin-horseradish peroxidase conjugates: I. Staining of principal cells and spermatozoa in the mouse

Several glycoconjugates are thought to bind spermatozoa as they pass through reproductive ducts. Paraffin sections of testis, ductuli efferentes, epididymis, and vas deferens of male mice were stained with ten different lectin-horseradish peroxidase conjugates to localize possible sites of synthesis and secretion of such glycoconjugates, based on the carbohydrate moieties in their constituent oligosaccharide side chains. Principal (columnar) cells lining the efferent ducts, germinal epithelium, and developing and maturing spermatozoa were examined with light microscopy. Staining of the Golgi and apical zones of cells was interpreted as evidence for synthesis and secretion of glycoconjugates. Principal cells synthesized and secreted glycoconjugates with sugar moieties as follows: sialic acid, all regions of the efferent ducts examined; the terminal disaccharide D-galactose- (β1 → 3) -N-acetyl-D-galactosamine, all regions of ducts except epididymis I; terminal α-D-galactosamine, some cells in epididymis III-V; N-acetyl-D-galactosamine, ductuli efferentes, epididymis I, II, and some cells in epididymis III-V; α-L-fucose, ductuli efferentes, vas deferens, and all regions of the epididymis except IV; N-glycosidic side chains, ductuli efferentes, vas deferens, and epididymis I, IV, and V. All of these sugar residues as well as N-acetyl-D-glucosamine were associated with the acrosomes and tails of spermatozoa throughout the ducts except for α-N-acetyl-D-galactosamine in epididymis I, and all occurred during one or more stages of spermiogenesis. The synthesis and secretion of glycoconjugates that bind to spermatozoa appear to involve more regions of the primary reproductive structures than was believed previously.     

Lysosomal integral membrane proteins exhibit region and cell type specific distribution in the epididymis of the adult rat

The epididymis, a post-testicular site required for maturation and storage of spermatozoa, is actively involved in exocytic and endocytic events, two phenomena likely to depend on the integrity of the lysosomal system. To study the lysosmal system of the epididymis, five monoclonal antibodies, previously characterized as recognizing five distinct lysosomal integral membrane proteins (LIMPs 1–5), were used as molecular probes of lysosome distribution in cells lining the epithelium. Immunocytochemical localization of LIMPs, using biotin-streptavidin immunoperoxidase methodology, was performed on frozen sections of adult rat epididymides and in cell cultures prepared from either the caput or cauda epididymis. In frozen sections, a heterogeneous distribution of the different LIMPs along the length of the epididymis was observed. For example, the distribution of LIMP 1 (35–50 K) was detected in all cells of the caput and quite dramatically in clear cells of the distal caput, corpus, and cauda epididymis, but specifically not in the principal cells of the distal caput, corpus, and cauda. In contrast, LIMP 2 (64–71 K) was present in all cells of the epidiyms, except clear cells. LIMPs 4 and 5 (93 K and 93 K) were detected in all epididymal cells, including the clear cells. Finally, whereas the regional and cell type distribution of LIMP 3 (74 K) in the epididymis was identical to that of LIMPs 4 and 5, the nature of the vesicles immunostained was distinct. In cultured cells, the general immunostaining patterns observed in vivo were maintained during the duration of the primary cultures for all five LIMPs. Our results begin to address the molecular heterogeneity of the lysosomal system along the lenght of the epidiymis, and may suggest in part a basis for underlying structural and functional characteristics of the epididymis leading to the sequential maturation of sperm.     

Morphological Changes of the Epididymis and Description of the Excurrent Ducts of Phrynops geoffroanus (Testudines: Chelidae) During the Reproductive Cycle

The seminal ducts (efferent ductule, epididymis, and deferent duct) in adults of Phrynops geoffroanus were examined using light microscopy. A series of tubules (efferent ductules) connect the testes to the epididymides. The efferent ductules are formed by a rete of small tubules of varying diameters, with simple columnar epithelium formed by the ciliated cells, nonciliated cells, and few basal cells. The epididymis is a simple, long and highly convoluted tubule that receives the efferent ductules throughout its extension. It is covered by a pseudostratified columnar epithelium with three cellular types: the principal cells, which are the most abundant, basal cells, and a small narrow cell. The histological differences in the epididymis region (cranial, medial, and caudal), as well as the differences in the epithelium throughout the reproductive cycle, are discussed. The deferent ducts consist of a low pseudostratified epithelium with two cellular types: the principal and basal cells. During the months analyzed, spermatozoa were stored in the epididymis, and deferent ducts were found. Anat Rec, 2010. © 2010 Wiley‐Liss, Inc.     

Histochemical evaluation of glycoconjugates in the male reproductive tract with lectin-horseradish peroxidase conjugates: II. Staining of ciliated cells,basal cells,flask cells,and clear cells in the mouse

Efferent reproductive ducts of male mice, including ductuli efferentes, epididymis, and vas deferens, were fixed and embedded in paraffin, and sections were stained with a battery of lectin-horseradish peroxidase conjugates to localize specific sugars or sugar sequences in glycoconjugates. Cilia and the apical surfaces of ciliated cells in the ductuli efferentes stained intensely with lectin specific for sialic acid and terminal α-N-acetyl-D-galactosamine. Flask cells and clear cells in the epididymis reacted positively and similarly with most lectins used, providing evidence that these cell types are related. In contrast, disparities in lectin staining suggest that flask cells and clear cells are a cell type distinct from principal cells. Basal cells were not present in the ductuli efferentes but formed a continuous layer in the epididymis and vas deferens. Basal cells contained oligosaccharides terminated by sialic acid and α-D-galactose and varying amounts of terminal β-D-galactose and α-N-acetyl-D-galactosamine. Basal cells also stained variably with lectins specific for the core region of complex type N-glycosidic side chains. The basal cells varied structurally, having long spinous apical processes approaching or reaching the lumen in region I of the epididymis and being low cuboidal or squamoid and lacking apical processes in epididymal regions II–V and in the vas deferens. The contiguous nature of the basal cells and the presence of glycoconjugates bearing terminal α-galactosyl residues in all basal cells suggest a possible role for these cells in a regulatory influence on transepithelial movement of fluid and/or ions in the epididymis and vas deferens.     

The differential absorptive activity of epithelial cells of the rat epididymus before and after castration.

Horseradish peroxidase introduced into the lumen of the rat epididymis was taken up by the columnar cells of the epithelium by five minutes and more so after longer periods. The apical cells and particularly the clear cells in the caput and cauda epididymis, respectively, showed significantly greater endocytotic activity than the principal cell in both locations. Within 14 days after castration, however, such differences in absorptive activity among the various cell types were essentially obscured because of increased endocytosis by the androgen-deficient principal cells. The results are discussed briefly in terms of the function of different epithelial cell types and secretory/absorptive activity in the epididymis.     

Permeability of the diaphragmatic mesothelium: The ultrastructural basis for “stomata”

The ultrastructure of the hamster efferent ducts and epididymis was studied and the results were correlated with previously published data on the composition of luminal fluid obtained by micropuncture. Samples of the efferent ducts and parts of the epididymis designated initial segment, caput, corpus, proximal cauda, distal cauda, and “epididymal vas” were prepared. The efferent ducts contained principal cells characterized by a profusion of apical vesicles and numerous very large vacuoles that were distributed throughout the cytoplasm. Ciliated cells had few vesicles and vacuoles. Occasional cells contained many particles resembling glycogen. In the epididymis, the following trends were observed. The height of the epithelium and the size of the principal cells declined from initial segment to distal cauda. Apical vesicles and vacuoles with a light content were extremely numerous in principal cells of the initial segment and decreased progressively in the more distal regions. In the initial segment, basal and perinuclear rough endoplasmic reticulum was abundant and was distended with a material that resembled newly synthesized protein. Further distally in the epididymis cisternae of the rough endoplasmic reticulum were narrow and contained little intracisternal material. Light cells containing many vesicles, vacuoles, and lysosome-like structures were very prominent in the caudal segments. The epithelium of the epididymal vas had features intermediate between cauda epididymidis and ductus deferens. The cytoplasmic droplet in luminal sperm began to migrate caudally between the caput and corpus epididymidis and reached the posterior extremity of the middle piece in the distal cauda. Some degenerating sperm were observed in the lumen of the distal segments of the epididymis. The abundance of cytoplasmic vesicles and vacuoles in principal cells of the efferent ducts and initial segment of the epididymis correlated with the site of greatest fluid absorption as determined by micropuncture studies, suggesting that these structures are involved in absorption of fluid from the lumen. Between the caput and distal cauda epididymal segments, where absorption of sodium and potassium but not of fluid occurred, there were few vesicles and vacuoles in principal cells, but the “light” cells were large and numerous and contained many vacuoles. The principal cells of the initial segment were best equipped with rough endoplasmic reticulum to synthesize a protein.     

Morphological and histochemical changes in the epididymis of hamsters (Mesocricetus auratus) subjected to short photoperiod

The morphological involution and histochemical changes of the Syrian hamster ( Mesocricetus auratus ) epididymis induced by a short light period were investigated. Under short-day conditions, the epididymis showed marked morphological changes including a decrease in luminal diameter, disappearance of spermatozoa, increase of interductal tissue, increase of intraepithelial lipofuscin deposits, the presence of phagolysosomes in the principal cells and macrophage-like cells, and a considerable modification of most clear cells. With lectin histochemistry changes were found in the glycoconjugates of principal cells of the regressed epididymis, either a decrease (PNA, WGA, HPA and DBA) or an increase (MAA) in the affinity of lectins to the Golgi area, or a decrease (HPA) or an increase (PNA) in lectin binding to stereocilia. Both morphological and histochemical results showed that, under this light condition, the cauda epididymidis presented the most prominent alterations, and that the epididymis showed increased absorptive activity and a decreased synthesis of glycoproteins. All these changes are probably due to the decrease in testosterone levels.     

实验性精索静脉曲张诱导青春期大鼠附睾细胞凋亡的研究

目的：研究实验性精索静脉曲张（EVC）大鼠附睾细胞凋亡及其显微、超微结构的变化。 方法： 采用青春期雄性Wistar大鼠复制左精索静脉曲张模型，以末端脱氧核糖核酸转移酶介导的dUTP缺口末端标记法（TUNEL）检测细胞凋亡,常规制作附睾体部光镜、电镜标本，观察附睾组织细胞形态变化。 结果： 实验组（VG）细胞凋亡率显著高于假手术组（SOG）（P<0.01），实验组左、右侧细胞凋亡率有差别，但无显著意义（P>0.05）。光镜下主要改变有附睾管萎缩，上皮细胞出现空泡化，上皮内晕、亮细胞数明显增多。电镜下主要表现为主细胞内溶酶体增多、变大，残余小体增加，内质网扩张，线粒体嵴模糊，高尔基复合体空泡化；核染色质致密，形成大小不等的团块，边集于核膜处；上皮细胞游离面微绒毛稀少，可见局灶性断裂和破坏。 结论： 青春期大鼠实验性精索静脉曲张可致附睾组织细胞凋亡过度，使其显微及超微结构明显改变，这些变化可能是影响精索静脉曲张患者生育能力的机制之一。     

Mast cells and ethanol consumption: interactions in the prostate, epididymis and testis of UChB rats

Citation
Mendes LO, Amorim JPA, Teixeira GR, Chuffa LGA, Fioruci BA, Pimentel TA, de Mello W Jr, Padovani CR, Pereira S, Martinez M, Pinheiro PFF, Oliani SM, Martinez FE. Mast cells and ethanol consumption: interactions in the prostate, epididymis and testis of UChB rats. Am J Reprod Immunol 2011; 66: 170–178 Problem Alcoholism has reached alarming proportions while fertility rates slowing in populations. The assessment of inflammatory effects with emphasis on the variation of the mast cells comparing ethanol chronic ingestion on reproductive organs deserves attention. Method of study The mast cells were investigated with light microscopy using toluidine blue to locate and count total mast cells and immunohistochemistry to identify the connective tissue mast cells (CTMC). Results The increase in total mast cells in the prostate, total and degranulated mast cells in epididymis of UChB rats was accompanied by a greater proportion of mucosal mast cells (MMC) in these organs. In addition, a lower incidence of degranulated mast cells was observed in epididymis of control rats. Conclusions Ethanol increases the number of total and degranulated mast cells in the prostate and epididymis, as well as associated with increasing MMC, and therefore, it could be leading to inflammation in these organs.     

Expression pattern of galectin-1 and galectin-3 in rat testes and epididymis during postnatal development

Galectins are a family of lectins-binding beta-galactosides involved in a variety of extracellular and intracellular processes, thereby contributing to homeostasis, cell adhesion, cellular turnover, and immunity. This study aimed to determine the localization and expression of galectin-1 (Gal-1) and galectin-3 (Gal-3) in the testis and epididymis of rats at postnatal [(prepubertal (day 5), pubertal (day 20), postpubertal (day 50) and mature (day 70)] periods by using immunohistochemistry and Western blotting. Gal-1 and Gal-3 were differentially expressed in different types of cells in the testis and epididymis during postnatal development. While we detected Gal-1 expression in some spermatogenic cells and Leydig cells in the testis, not in the epididymal epithelium, Gal-3 was expressed in Sertoli cells, peritubular myoid cells, Leydig cells, smooth muscles and interstitial CD68-positive macrophages. Epithelial cells of the corpus and cauda epididymis showed an intense Gal-3 expression. Gal-1 expression was higher in the testis than in the epididymis on days 50 and 70. The expression of Gal-3 in the testis increased from the prepubertal to mature period. While the expression difference of Gal-3 was not statistically significant in the testis and epididymis until puberty, Gal-3 expression in the postpubertal and mature periods was higher in the epididymis. The expression of Gal-3 in the corpus and cauda epididymis was higher than that in the caput epididymis. In conclusion, our findings suggest that puberty has potential regulatory effect on the expression of galectins in testis and epididymis of rats. Gal-1 and 3 may play a role in the development of the reproductive system and the preservation of the immune-privileged environment in the testis, due to their pro-apoptotic and anti-apoptotic functions. The presence of intense expression of Gal-3 in the corpus and cauda epididymis may contribute to the maturation and storage of spermatozoa.     

Cell- and region-specific expression of sugar chains in the mouse epididymal epithelium using lectin histochemistry combined with immunohistochemistry

To understand the cytochemical properties of epididymal epithelial cells, the characteristics of glycoconjugates in the mouse epididymis were examined using the technique of lectin histochemistry combined with immunohistochemistry. Characteristic staining patterns depending on the type of lectins were observed in the epididymal epithelium. Principal cells expressed N-acetyl-D-galactosamine (GalNAc), N-acetyl-D-glucosamine (GlcNAc), and Fucose in the proximal region of the epididymis and Mannose, Glucose, and Galactose in the distal region of the epididymis. Basal cells expressed Mannose, Glucose, Galactose, and GlcNAc in the proximal region and Galactose in the distal region. On the other hand, clear cells expressed various sugar residues and differences among regions were not observed. Interestingly, principal cells, clear cells, and basal cells specifically reacted with Ulex Europaeus-Agglutinin I (UEA-I lectin), Maackia Amurensis-Lectin I (MAL-I lectin), and Griffonia simplicifolia Lectin I-B4 (GS-I B lectin), respectively. These findings indicate that the selectivity in lectin reactivity for distinct cell types and segment-dependent staining in the epididymis may be related to cellular and regional differences in function. Furthermore, because some lectins stain particular cells or cellular compartments selectively, these lectins could be useful markers for histopathological evaluation of diseases or diagnosis of male infertility.     

Organization of tubules in the human caput epididymidis and the ultrastructure of their epithelia

The structure of the human caput epididymidis was examined by gross morphological and light and electron microscopic techniques. There were at least seven types of tubules, each characterized by a different epithelium. These tubules were connected with one another by at least eight types of junctions to form a network. Most of the caput epididymidis was composed of efferent ducts. Within these, five types of tubules, each with a different ciliated epithelium, were found in different regions; and four types of junctions between the efferent ducts and the epididymal tubule were observed. The efferent ducts left the testis, initially as parallel straight tubules containing both ciliated and non-ciliated cells in an epithelium of irregular height. Each efferent duct then coiled tortuously into lobules that folded over one another. These efferent ducts then branched out as thin tubules to join a network of dark tubules which were lined by a regular epithelium containing prominently vacuolated, nonciliated cells. These tubules anastomosed via common cavities characterized by a ciliated cuboidal epithelium and sometimes joined tubules exhibiting a non-vacuolated ciliated epithelium. The latter, as well as typical efferent ducts, made connection with the epididymis proper in both end-to-end and end-to-side junctions. In the more distal junctions with the epididymis, the efferent ducts joined to a transitional epididymal ductule before joining to the side of the epididymis proper. Post-junctional epithelia in the beginning of the epididymis occasionally contained patches of cells characteristic of efferent ducts. Tall cells with long stereocilia constituted a discontinuous “initial segment”-like region of the epididymis. This is the most detailed study so far of the epithelia and the tubule organization in the caput epididymidis of any species, and most of the results are reported for the first time for the human. Although the pattern of the tubule network resembles that of some domestic species, the rich variety of epithelia has not been appreciated before.     

3 beta-Hydroxysteroid-dehydrogenases of the testis and epididymis of the Peking duck (Anas platyrhynchos L.)

This study is concerned with the histochemical activity of hydrosteroid dehydrogenases (HSD), glucose-6-phosphate dehydrogenase (G6PDH), and 6-phosphogluconate dehydrogenase (6PGDH) in testis and epididymis of 20 male Pekin ducks. Various 3 beta-HSD, the G6PDH, and the 6PGDH were localized in these organs. 3 beta-etiocholane was much quicker utilized by the 3 beta-HSD than epiandrosterone, dehydroepiandrosterone, and androstendiol. The capability of testis Leydig cells of utilizing the 3 beta-etiocholane was higher in winter than in summer. The G6PDH and the 6PGDH in the Leydig cells and basal cell layer of the tubuli seminiferi contorti were active throughout the year. There was weak 3 beta-HSD activity in the epithelia of the ductuli efferentes distales and in the ductus epididymidis from March to June. The G6PDH and the 6PGDH showed positive reaction in the rete testis and ductuli efferentes proximales throughout the year. The 3 beta-HSD activity in testis and epididymis is saisonally different: In summer, there is no 3 beta-HSD activity in both organs. In winter, the 3 beta-HSD is demonstrable in the testis but not in the epididymis. In spring, there is 3 beta-HSD activity in the epididymis but not in the testis.     

The initial segment of the rat epididymis is able to uptake immature germ cells shed by testicular damage

The initial segment of the caput epididymidis, the most proximal part of the rat epididymis, has specific functional characteristics. In the present study, the behavior of the epididymal epithelium from this region was evaluated after the exposure to a massive number of immature germ cells in the luminal fluid. The experimental release of immature germ cells from the seminiferous tubules was performed by injecting anti-microtubule compounds into the rete testis and the lumen of seminiferous tubules. Twenty-four hours after nocodazole or colchicine administration, a massive phagocytosis of immature spermatogenic cells, recognized as acrosin-positive structures, was easily observed in the epithelium of the initial segment of the epididymis assessed by light and electron microscopy. Immature germ cells were engulfed by epithelial cells, where most of them were found as cell debris at different stages of degradation. No signs of inflammation were observed either in the lumen or in the interstitium. The phagocytosis of immature germ cells was restricted to the epithelium of the initial segment of the epididymis, suggesting a role for this segment as the first selective barrier for the exclusion of abnormal gametes along the male genital tract.     

A Study of Effect of Progesterone on Epididymis in Albino Rats

Progesterone, an anti androgen compound that recently receiving attention for potential use as male contraceptive and for other medical purposes, such as treatment of prostate diseases. In the present study adult male albino rats were administered the progesterone for 15 &; 30 days and the histology and fine structures of the epididymis were studied. After treatments for 15 and 30 days sperms were found to greatly reduce in number from lumen of caput epididymis, middle segment and cauda epididymis, of severely affected specimen. The epithelium was tall and the light cells were large and distended with many dense bodies resembling lysosome (Loving &; Flickinger 1976)1. The lumen was filled with scanty sperm and debris, which appears to be derived from germ cells. It is suggested that the light cells of epididymal epithelium may have a role in clearing the lumen in which they are particularly large and numerous.The aim of present study is to determine the effect of progesterone on the structure of target cells of epididymis normally stimulated by androgens and further correlate the findings in light of previous studies, to draw the significant conclusion. The study showed that the progesterone have intense inhibitory effect on the epididymis. The degenerative histological findings are found in form of reduce number of spermatozoa, debris of cell mass &; reduce epithelial cells height. These changes may have an important role in the anti fertility effect of progesterone.     

层黏连蛋白受体在小鼠睾丸和附睾中的表达

目的 探讨层黏连蛋白受体(LAMR1)在小鼠睾丸和附睾中的表达.方法收集3只正常成年昆明小鼠睾丸和附睾.采用原位杂交和免疫组织化学方法,检测LAMR1 mRNA及蛋白在成年小鼠睾丸和附睾中的分布.结果 LAMR1 mRNA在附睾头和附睾尾表达最强;在睾丸生精细胞中也有表达.免疫组织化学结果显示,LAMR1蛋白从附睾头到...     

癫痫对雄性大鼠睾丸及附睾组织病理学和超微结构的影响

目的：探讨癫痫发作对雄性Wistar大鼠睾丸及附睾组织病理和超微结构的影响。方法：运用氯化锂一匹罗卡品建立Wistar雄性大鼠癫痫模型,取睾丸、附睾分别制片观察组织病理及超微结构的形态学改变。结果：光镜观察下,A组（造模成功组）及B组（造模不成功组）大鼠睾丸生精小管内各级生精细胞排列基本整齐,结构未见明显紊乱现象,但生精细胞层次呈不同程度的减少,睾丸生精小管管腔内精子数目减少,可见坏死脱落的生精细胞,睾丸间质结构缺如,间质细胞明显减少。附睾中,A组管壁柱状细胞,基细胞层次清晰,结构整齐,微绒毛排列整齐,但管腔中精子数目明显减少,有较多的非精子细胞成分。B组附睾管腔中精子数目未见明显下降,有时可见散在的生精细胞。电镜观察下,A组大鼠睾丸生精细胞细胞核明显畸形,线粒体肿胀,线粒体膜仍然完整,但脊消失,粗面内质网肿胀明显,精子头部细胞核清晰,顶体形态不规则,尾部“9＊2＋2”结构整齐,周围包绕的线粒体鞘明显肿胀,线粒体数目明显减少。B组中仍可见上述不同程度的损害表现,附睾中,A组及B组均可见处于同一层面的主细胞,细胞核未见明显异常,核周围可见大量溶酶体,同时核周内质网均处于明显肿胀状态。C组（正常对照组）鼠睾丸及附睾切片的光镜、电镜表现皆正常。结论：癫痫不同程度的发作可引起大鼠睾丸及附睾组织病理和超微结构不同程度的改变,进而造成了雄性Wistar大鼠生殖系统相关指标的改变。