Estrogen receptor in the ductuli efferentes, epididymis, and testis of rhesus and cynomolgus macaques

We obtained the testes, ductuli efferentes, and epididymides from adult rhesus and cynomolgus macaques and examined these tissues for estrogen receptors (ER) with immunocytochemistry (ICC) and a sucrose gradient assay. Both techniques employed monoclonal antibodies prepared against ER, and both showed that high concentrations of ER were present OFFy in the ductuli efferentes. Moreover, all specific staining was confined to the nuclei of the nonciliated, absorptive epithelial cells. The quantity of salt-extractable ER in the ductuli efferentes (834 +/- 161 [SEM] fmol/mg DNA [n = 8]) did not differ significantly from the amounts measured with the identical assay in oviducts and endometrium of estrogenized female macaques. Testes and epididymides of macaques had no specific staining by ICC and barely detectable amounts by biochemical analysis (7 +/- 4 [n = 3], 8 +/- 2 [n = 5], 33 +/- 16 [n = 3], and 6 +/- 3 [n = 8] fmol/mg DNA for testis and caput, corpus, and cauda epididymis, respectively). The functional significance of the high levels of ER in the ductuli efferentes of macaques remains to be determined.     

Immunohistochemical investigation of different cytokeratins and vimentin in the human epididymis from the fetal period up to adulthood

Summary The anatomical distribution of cytokeratins and vimentin was investigated by means of immunohistochemistry in the human epididymis. Epithelial cells of the ductuli efferentes and the corpus epididymidis were positive for cytokeratins and vimentin. The expression of epithelial vimentin decreased toward the cauda epididymidis, whereas cytokeratins remained unchanged. The epithelium of the ductus deferens was negative when antibodies against vimentin were used. With monoclonal antibodies to individual cytokeratins, the presence of cytokeratins 7, 8, 18, and 19 was demonstrated histochemically throughout the epithelium of the epididymis. Monoclonal antibodies specific for cytokeratin 17 allowed immunohistochemical differentiation between the ductuli efferentes and the ductus epididymidis.     

Morphology of the epithelium of the extratesticular rete testis, ductuli efferentes and ductus epididymidis of the adult male rabbit

The fine structure of the epithelium lining the extratesticular rete testis, ductuli efferentes and ductus epididymidis of the rabbit has been investigated. In the ductuli efferentes the epithelium is composed of two cell types, principal cells and ciliated cells. The latter type is distinguished from principal cells by the presence of cilia projecting into the lumen and the position of the nucleus in the apical half of the cell. Principal cells in this segment are characterized by micropinocytotic vesicles on the surface plasma membrane and a variety of small dense bodies scattered throughout the cytoplasm. In the ductus epididymidis basal cells replace ciliated cells as the second cell type, but differences between various segments of the epididymis are related to the fine structure of the principal cells. In the proximal caput epididymidis (Nicander's region 1) the principal cells are tall with long microvilli. They typically contain a small Golgi apparatus and a cluster of dense bodies adjacent to the nucleus. In the distal caput epididymidis (Nicander's regions 2-5) the apical cytoplasm of principal cells is filled with numerous micropinocytotic vesicles and large multivesicular bodies; these features are interpreted as signs of absorptive activity. The multivesicular bodies are absent from the cytoplasm of principal cells in the corpus epididymidis (Nicander's region 6) and, instead, numerous elements of smooth endoplasmic reticulum, a large Golgi apparatus, lipid droplets and dense bodies characterize principal cells in this segment. Towards the proximal cauda epididymidis (Nicander's region 7), the number of dense bodies (lysosomes) in the cytoplasm increases considerably. In the globose cauda (Nicander's region 8), the principal cells are reduced in height, and in addition to the features described in region 7, are characterized by a concentric array of rough endoplasmic reticulum in the basal cytoplasm. These observations are discussed in relation to the role of the epididymis in promoting the maturation and survival of spermatozoa.     

Intraepithelial lymphocytes and macrophages in the human epididymis

The epididymides from 7 young adults and 5 older patients with prostatic cancer were investigated by means of light and electron microscopy. The existence of lymphocytes and macrophages between epithelial cells of various segments of the ductuli efferentes and the ductus epididymidis is proven, and the distribution and morphology of these cells are described. A possible relationship between these two intraepithelial cell types, as well as their presumptive role with regard to an immunological barrier-function are discussed.     

Transport of IgG across the blood-luminal barrier of the male reproductive tract of the rat and the effect of estradiol administration on reabsorption of fluid and IgG by the epididymal ducts

In rats immunized systemically with tetanus toxoid the concentration of specific anti-tetanus-toxoid-specific IgG in fluid from the rete testis and cauda epididymidis were respectively 0.6% and 1.4% the concentration in blood serum. The extratesticular duct system reabsorbed 97% of the IgG and 99% of the fluid leaving the rete, but estradiol administration affected the site of reabsorption. In untreated rats, the ductuli efferentes reabsorbed 94% of the IgG and 96% of the fluid leaving the rete, whereas estradiol-treated rats reabsorbed 83% of the IgG and 86% of the fluid, and the ductus epididymidis fully compensated for these different effects of estradiol on the ductuli efferentes. The concentrations of IgG in secretions of the seminal vesicles and prostate gland were lower (0.1% and 0.3% respectively of the titers in blood serum) than in fluids from the extratesticular ducts, and were not affected by the administration of estradiol. RT-PCR showed that Fcgrt (neonatal Fc receptor, also known as FcRn) is expressed in the reproductive ducts, where IgG is probably transported across epithelium, being particularly strong in the ductuli efferentes (where most IgG was reabsorbed) and distal caput epididymidis. It is concluded that IgG enters the rete testis and is concentrated only 2.5-fold along the extratesticular duct system, unlike spermatozoa, which are concentrated 95-fold. Further, the ductus epididymidis can recognize and compensate for changes in function of the ductuli efferentes.     

Phagocytosis of spermatozoa by the epithelial cells of the ductuli efferentes after epididymal obstruction in the rat.

The effects of ligation of an isolated loop of the ductus epididymidis in the region of the caput were compared with those of the exfoliative lesion of the epididymis that follows administration of alpha-chlorhydrin to rats. Electron micrographs of the ductuli efferentes in both cases revealed early phagocytosis of apparently normal spermatozoa by the epithelial cells, followed at later intervals by invasion of macrophages and intraluminal phagocytosis. It is concluded that epithelial spermiophagy is a consequence of obstruction, whether mechanically or chemically induced.     

Protein gene product 9.5 and ubiquitin immunoreactivities in rat epididymis epithelium

A quantitative immunohistochemical study was performed of the distribution of protein gene product 9.5 (PGP, a soluble protein localized in neurons and neuroendocrine cells as well as in some non-nervous cells) and ubiquitin along the rat epididymis. In the ductuli efferentes, PGP immunoreaction was observed in the whole cytoplasm of some columnar cells; a smaller number of columnar cells showed ubiquitin immunoreactivity with limited apical and basal cytoplasmic localization. In the proximal caput epididymidis, the whole cytoplasm of all columnar cells showed PGP immunoreactivity, ubiquitin immunostaining was negative in this region. In the middle and distal caput epididymidis and the distal cauda, the apical cytoplasm of some columnar cells and the whole cytoplasm of some basal cells showed immunoreactivity to PGP. In these regions, immunoreactivity to ubiquitin was positive in the supranuclear cytoplasm of some columnar cells but not in the basal cells. No immunoreactivity to PGP or ubiquitin was detected in the corpus epididymis and the proximal cauda. Double immunostaining revealed that all the epididymal ubiquitin immunoreactive cells were also PGP immunoreactive, whereas most PGP immunoreactive cells did not immunoreact to ubiquitin. In ubiquitin-PGP immunoreactive cells, the site of the PGP immunoreaction differed from that of the ubiquitin immunoreaction. PGP-ubiquitin immunoreactive cells also seemed to be immunoreactive to anti-AE1/AE3 keratin antibodies. The spermatozoal heads were immunoreactive to PGP antibodies in the epididymal regions from proximal caput to distal cauda but not in the ductuli efferentes. The findings suggest that non-ubiquitinated PGP immunoreactive proteins are secreted in the epididymis, mainly in the proximal caput, and attach to spermatozoa.     

Morphometric studies on rat epididymis in the course of postnatal development (computerised image analysis)

The studies were aimed at histological and morphometric analysis of postnatal development of epididymis in rats aged 1, 5, 10, 20, 28, 35, 45 and 59 days. Diameter of ductuli efferentes and of ductus epididymidis, height of epithelium, section area fractions occupied by the epithelium and by the sublayer were estimated (using MultiScan software) and serum testosterone levels were measured. The results documented a stepwise development of the epididymis, in three distinguishable stages.     

Changes in the composition of the excurrent duct system of the rat testis during postnatal development.

The gross composition of the testicular excurrent duct system of the rat was examined and compared along the length of the duct and with samples of testis, bladder and liver. Changes in composition with age were examined by analysing tissue from animals at postnatal ages of 19, 36, 48, 60, 90 and 120 days. In adult animals, testicular tissue was characterized by having the lowest dry weight, accompanied by low levels of total protein, lipid, RNA and glycogen; DNA, phospholipids and sialic acid were at levels similar to other tissues. A high proportion of the total protein was soluble. The ductuli efferentes plus initial segment of the epididymis were characterized by high levels of total lipid. The caput epididymidis contained a low level of total protein but a high level of acid-soluble phosphorus. The cauda epididymidis had a low dry weight and low levels of total protein, soluble protein, and lipid, but high levels of acid-soluble phosphorus, DNA and sialic acid. The ductus deferens contained small amounts of RNA and DNA but had a high dry weight, high total protein, soluble protein and glycogen. Several trends were apparent with increasing age. Dry weight increased in the ductuli efferentes plus initial segment, whilst total protein decreased in the caput and cauda epididymidis. Total lipid increased in the ductuli efferentes plus initial segment and acid-soluble phosphorus and sialic acid increased in all other segments of the excurrent duct system. In all segments the content of RNA and DNA decreased as the animals matured. The concentration of calcium and magnesium in the excurrent duct system was not significantly different from those levels found in the liver. High levels of spermine and spermidine were confirmed in the prostate, and were also detected in the testis, caput epididymidis and cauda epididymidis, but at a much lower concentration.     

A study of the effect of high concentrations of fluoride on the reproductive organs of male rabbits, using light and scanning electron microscopy

Fluoride was orally administered to rabbits at 10 mg NaF/kg body weight for 18 or 29 months. The animals were then killed and the structure of the testis, epididymis and vas deferens studied under light and scanning electron microscopes. In animals treated for 29 months, the spermatogenic cells in the seminiferous tubules were disrupted, degenerated and devoid of spermatozoa. In animals treated for 18 or 29 months, loss of cilia on the epithelial cells lining the lumen of the ductuli efferentes of the caput epididymidis and of stereocilia on the epithelial cells lining the lumen of the vas deferens was observed. In some regions of the epithelial lining of the lumen of the ductuli efferentes and vas deferens, the boundaries of the cells were not clear and appeared to be peeled off. Mucus droplets were abundant in the vas deferens of control animals, but absent in both the treated groups. Spermatogenesis ceased only in animals treated for 29 months. The difference in the structural changes observed in the testes of the 2 treated groups may have been due to the blood-testis barrier. It is concluded that ingestion of high concentrations of fluoride has harmful effects on the male reproductive system.     

Estrogen receptor alpha has a functional role in the mouse rete testis and efferent ductules

Previous studies of the estrogen receptor-alpha knockout (alpha ERKO) in the male mouse demonstrate that the rete testis and efferent ductules are targets of estrogen. Because the alpha ERKO mouse lacks a functional estrogen receptor alpha (ER alpha) throughout development, it was not known whether the morphological and physiological abnormalities observed in the alpha ERKO male were due to developmental defects or to dysfunctions concurrent with the lack of ER alpha in the tissue. This study was designed to determine if treatment of normal wild-type (WT) mice with the pure antiestrogen, ICI 182,780, (ICI) could reproduce the morphological characteristics seen in alpha ERKO mice. Thirty-day-old male mice were treated for 35 days with either castor oil or ICI. Age-equivalent alpha ERKO mice were used for comparison. Light microscopic examinations of the reproductive tracts revealed dramatic changes in the efferent ductules of treated mice: a 1.7-fold increase in luminal diameter, a 56% reduction in epithelial cell height, a 60% reduction in brush boarder height of nonciliated cells, and an apparent reduction of the number of observable lysosomes and endocytotic vesicles. Testes of ICI-treated mice showed swollen rete testes area (6.5 times larger than control) and a 65% reduction in rete testis epithelium height. However, there were no significant changes in body and testis weights. These results indicate that ER blockage with ICI in WT mice results in morphological changes of the efferent ductules resembling those seen in alpha ERKO siblings of the same age. Based on this study, we conclude that ER alpha has a functional role in the mouse reproductive tract and the aberrant morphology observed in the efferent ductules of the alpha ERKO mouse is likely the result of a concurrent response to the lack of functional ER alpha, and not solely due to the lack of ER alpha during early developmental times.     

The epididymal region of the Japanese quail (Coturnix coturnix japonica).

The epididymal region of the Japanese quail was studied histologically. The organ consists of the extratesticular portion of the rete testis, the ductuli efferentes proximales and distales, the ducti conjugentes and ductus epididymidis. Distinct tubuli recti link the seminiferous tubules with the rete testis. The non-ciliated cells of the ductuli efferentes proximales and distales show, between them, certain internal structural differences which were highlighted. In 40% of the birds, the ductus deferens showed dark-grey pigments, regarded as melanin. The epididymal region was generally similar in structure to that of the domestic fowl, turkey and duck.     

Changes in protein composition of the luminal fluids along the epididymis of the tammar, Macropus eugenii

Micropuncture samples of luminal fluid were collected from the rete testis and along the epididymis. Quantitative analyses showed that the ductuli efferentes reabsorb about half the protein leaving the testis. Considerable protein is secreted by the caput epididymidis (initial segment) and there is a net loss of protein from the corpus and cauda epididymidis. Denatured, polyacrylamide gel electrophoresis showed that there are 5 proteins in rete testis fluid which are not present in blood (Mr of 14,700, 22,800, 24,100, 43,000 and 44,800). One of these proteins (Mr 14,700) is lost from plasma in the ductuli efferentes and 2 (Mr 43,200 and 44,800) are lost in the corpus epididymidis. Twelve proteins appear in the epididymal plasma and are not present in rete testis fluid or blood: 6 appear in the caput epididymidis (Mr 30,000, 31,000, 32,300, 17,400, 18,700 and 21,400), 3 in the corpus epididymidis (Mr 12,800, 39,800 and 90,600) and 3 in the cauda epididymidis (Mr 10,900, 56,300 and 63,000). A protein with the same molecular weight as a blood protein (149,500) accumulates in the corpus and cauda epididymidis. None of the samples of luminal fluid contained particulate matter other than spermatozoa, indicating that the tammar is a useful animal for micropuncture studies.     

Structure and functions of the genital ducts of the male Port Jackson shark,Heterodontus portusjacksoni

Synopsis The genital ducts ofHeterodontus portusjacksoni consist of the sperm carrying ducts (the rete testis, ductuli efferentes, and initial and terminal segments of the ductus epididymidis) and the Leydig glands (anterior opisthonephros). The ducts are lined by a ciliated epithelium which maintains a barrier to the transport of solute between blood and the lumen of the duct. Spermatozoa, Sertoli cell bodies, Sertoli cell cytoplasts and cellular debris are released from spermatocysts into the longitudinal canal of the rete testis. However, only the Sertoli cell cytoplasts persist throughout the sperm ducts. The epithelia lining the initial segment of the ductus epididymidis and secretory tubules of the Leydig glands are specialized for protein secretion and (particularly the Leydig glands) must be the main source of luminal protein in the ductus epididymidis. The epithelium lining the terminal segment of the ductus epididymidis also secretes protein, reabsorbs fluid and sodium, and may carry out heterophagic digestion. Spermatozoa develop the capacity for motility in the extratesticular sperm ducts, but do not undergo structural changes. However, they form spherical bundles in the terminal segment of the ductus epididymidis. It is suggested that the reduction in ratio of sodium:potassium from 48:8 in the ductuli efferentes to 3:4 in the distal end of the terminal segment of the ductus epididymidis may favour sperm survival.     

Histological localization of hydrolases in the epididymis of the dog]

Localization and activity of five hydrolases (alkaline phosphatase, adenosine triphosphatase, acid phosphatase, nonspecific esterase and leucylamino-peptidase) were evaluated histochemically in the epididymides of mature dogs. In the ductuli efferentes, cilia and apical parts of the epithelial cells displayed high activity of alkaline phosphatase and adenosine triphosphatase. Strong activity of acid phosphatase, nonspecific esterase and leucylamino-peptidase was present in the basal and supranuclear zones of the epithelium of the ductuli efferentes. Stereocilia of all three segments of the ductus epididymidis showed a high activity of alkaline phosphatase. Positive adenosine triphosphatase reaction was confined to the stereocilia of the initial segment. A complex pattern of acid phosphatase activity was observed in the middle segment. The subdivision of the middle segment in four subsegments was therefore suggested. In the epithelium of the initial segment only a few nonspecific esterase-positive cells were seen. The infranuclear and basal areas of the epithelium in the middle segment and the supranuclear zone of the terminal segment displayed distinct nonspecific esterase activity. The possible contribution of the hydrolases to the function of the epididymis is discussed.     

Transformation of the Mesonephros into Epididymis in the Cock: Fate of Collecting and Secretory Tubule Antigens

By means of purified rabbit anti-adult chicken kidney antibodies two types of antigens have been identified in the mesonephros: one (PS), localized in the cells of the proximal secretory tubules, the other (CT) characteristic of the collecting segments.
During the conversion of certain mesonephric tubules into epididymal ductules in the male, the CT-antigen disappears, whereas the PS-antigen not only persists but can also be detected in the cells of the parietal layers of Bowman's capsules as they transform before connecting with the rete testis. In the adult cock, the PS-antigen is found at the apical part of the cells of the ductuli efferentes, whereas the ductuli conjugentes and ductus epididymidis are negative.
In the female, the PS-antigen is also found to persist in the cells of the proximal segment and to appear in the cells of the parietal layers of Bowman's capsules. However, between 2 and 3 months after hatching it disappears, although perfectly well preserved tubuli of mesonephric origin can be observed at much later stages.
In the epididymal tubuli of the sexually mature cock, large sheets of the epithelial walls are seen to detach and to fall into the lumina where they disintegrate. The cellular contents thus released mix with the mass of the spermatozoa. Arguments tending to exclude a fixation artifact are presented and the possibility that the observation reflects a cyclic physiological process is discussed.     

Ultrastructure of the reproductive system of the black swamp snake (Seminatrix pygaea). VI. Anterior testicular ducts and their nomenclature

In this study, the anterior testicular ducts of the North American natricine snake Seminatrix pygaea are described using light and electron microscopy. From the seminiferous tubules, the rete testis passes into the epididymal sheath, a structure along the medial border of the testis heavily invested with collagen fibers. The rete testis consists of simple, nonciliated cuboidal epithelium (principal cells). The intratesticular ducts of the rete testis are narrow (50–70 μm) at their junction with the seminiferous tubules, widen (80–100 μm) as they extend extratesticularly, and divide into smaller branches as they anastomose with the next tubules, the ductuli efferentes. The ductuli efferentes are lined by simple cuboidal epithelium but possess nonciliated principal cells as well as ciliated cells. These are the only ducts in the male reproductive system with ciliated cells. The ductuli efferentes are narrow (25–45 μm), divide into numerous branches, and are highly convoluted. The ductus epididymis is the largest duct in diameter (240–330 μm), and the diameter widens and the epithelium thins posteriorly. The ductus epididymis is lined by nonciliated, columnar principal cells and basal cells. No regional differences in the ductus epididymis are apparent. Ultrastructural evidence suggests that all of the nonciliated principal cells in each of the anterior testicular ducts function in both absorption and secretion. Absorption occurs via small endocytic vesicles, some of which appear coated. Secretion is by a constitutive pathway in which small vesicles and a flocculent material are released via a merocrine process or through the formation of apocrine blebs. The secretory product is a glycoprotein. Overall, the characteristics of the anterior testicular ducts of this snake are concordant with those of other amniotes, and the traditional names used for snakes are changed to conform with those used for other sauropsids and mammals. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Ultrastructural and cytochemical study on the epithelium lining ductuli efferentes in Equus asinus

The ultrastructure and cytochemistry of the epithelium lining ductuli efferentes in adult Equus asinus was studied. The epithelium is constituted of randomly distributed ciliated and non-ciliated cells extending from the basement membrane to the lumen. A third cellular type appearing to be in a degenerating state may also occur with variable frequency and is basally located. Mainly "non-ciliated cells" display ultrastructural and cytochemical features which can be related to strong resorptive activity; lysosomes and pigment granules, lipofuscinic in nature, are also present. Pigment masses exhibiting the same morphological and histochemical features fill the cytoplasm of degenerating cells. The results are compared with those obtained by previous authors in other Mammals and in Birds and are related to the functions commonly attributed to ductuli efferentes. The origin of degenerating cells, which are peculiar of Equidae, is discussed.     

国人睾丸、附睾和输精管的氧化还原酶的组织化学观察

本文收集了１９—３８岁国人正常男性新鲜睾丸、附睾和输精管１３例,进行了氧化还原酶组织化学染色、光镜定位及定性观察。结果表明：睾丸曲细精管和输出小管上皮的ＧＤＨ,ＮＡＤＨＤ,ＮＡＤＰＨＤ,ＳＤＨ,ＧＰＤＨ,ＩＣＤＨ,ＭＤＨ,ＬＤＨ和Ｇ－６－ＰＤＨ９种酶;睾丸间质细胞和附睾管上皮的ＮＡＤＨＤ,ＮＡＤＰＨＤ,ＳＤＨ,ＩＣＤＨ,ＭＤＨ,ＧＤＨ,ＬＤＨ和Ｇ－６－ＰＤＨ８种酶;输精管的ＮＡＤＨＤ,ＮＡＤＰＨＤ,ＩＣＤＨ和ＧＤＨ４种酶的酶活性呈强阳性或极强阳性。提示输出小管和头部附睾管含有的多种氧化还原酶对精子功能成熟有极重要作用。