Epididymal Duct Research Articles

Ontogeny of the testicular excurrent duct system of male Japanese quail (Coturnix japonica): A histological, ultrastructural, and histometric study.

The testicular excurrent duct system undergoes several physiological and morphological changes during the reproductive stage or breeding season in mammals, birds, and reptiles. Studies on normal age-related histomorphological changes in the excurrent duct system of Japanese quails (Coturnix japonica) remain unreported, despite the extensive use of this bird as an avian model in research studies. The current study investigated the histological, ultrastructural, and histometric changes in the testicular excurrent duct system of the Japanese quail during three reproductive stages, namely prepubertal, pubertal, and adult. Simple squamous to low cuboidal cells formed the epithelia of the rete testis in prepubertal and pubertal birds, while in adult birds the lining was low cuboidal to cuboidal. In pubertal and adult birds, the nonciliated Type I epithelial cells of the proximal efferent duct displayed a subapical endocytotic apparatus comprising coated pits, coated apical tubules, and endosomes. There was a significant increase (p ≤ .001) in epithelial heights of all ducts of the excurrent duct system in the mature, sexually active, adult birds when compared to the other age groups. The luminal and tubular diameters, and the cross-sectional areas of efferent ducts and the epididymal duct unit increased significantly (p ≤ .001) with age. It is concluded that the morphology and morphometry of the excurrent ducts of the testis of the Japanese quail change as birds mature.

Mechanisms underlying spontaneous phasic contractions and sympathetic control of smooth muscle in the rat caudal epididymis.

Here we investigate mechanisms underlying spontaneous phasic contractions (SPCs) and sympathetic control of contractility in the rat epididymis, a long tubular duct involved in transportation and maturation of sperm. Longitudinal contractions of short segments (~ 1.5mm) of rat proximal and distal caudal epididymal duct were measured + / - nerve stimulation. The extent of sympathetic innervation of these duct regions was determined by immunohistochemistry. Proximal caudal duct segments (150-300μm dia.) exhibited SPCs, while distal segments (350-500μm) were quiescent in ~ 80% of preparations. SPC amplitude and frequency were reduced by the L-type voltage-dependent Ca2+ channel (LVDCC) blocker nifedipine (1μM), with the T-type voltage-dependent Ca2+ channel (TVDCC) blocker ML218 (1μM) specifically decreasing SPC frequency. SPCs were inhibited upon blockade of the SR/ER Ca2+-ATPase (CPA 10μM). SPCs were also inhibited by caffeine (1μM), 2-APB (100μM), niflumic acid (100μM), or by lowering extracellular [Cl-] from 134.4 to 12.4mM but not by ryanodine (25μM) or tetracaine (100μM). Electrical field stimulation (EFS) at 2Hz for 60s caused a sustained α1-adrenoceptor-sensitive contraction in distal segments and enhanced and/or induced α2-adrenoceptor-sensitive oscillatory phasic contractions in proximal and distal segments, the latter mimicked by application of the α2-adrenoceptor agonist clonidine. We hypothesise that SPCs in the proximal cauda are triggered by pacemaker mechanisms involving rhythmic IP3 receptor-operated SR/ER store Ca2+ release and resultant activation of CaCC with TVDCCs and possibly LVDCCs subserving in this process. Sympathetic nerve-released noradrenaline induces α2-adrenoceptor-mediated phasic contractions in the proximal and distal cauda. These findings provide new pharmacological targets for male infertility and contraception.

Effects of Diabetes Mellitus on the Reproductive System of Adult Male Mice After One Cycle of Spermatogenesis

Background: Diabetes mellitus is a disease characterized by chronic hyperglycemia. Poor management of hyperglycemia leads to complications that can reduce quality of life. Diabetes mellitus has been associated with damage to the reproductive system. The present study examines effects of diabetes mellitus on the reproductive system in alloxan-induced diabetic mice after one cycle of spermatogenesis. Materials and Methods: Twelve (12) mice divided into "control" and "diabetic" groups of six (6) animals each were used. Diabetes was induced in adult male mice by intraperitoneal injection with a single dose of 220 mg/kg body weight of alloxan. After 40 days, sperm density, morphology, and motility were assessed by standard methods. Serum levels of testosterone, FSH, and LH were measured. In addition, testes and epididymis were removed for histological study. Results: Highly significant (P < 0.001) reductions in serum testosterone, FSH, and LH levels, as well as normal and motile sperm counts of 21% and 34% respectively were observed in the diabetic group. The control group had three times more sperm than the diabetic group. The histological increased inter-tubular spaces, loss of interstitial tissue, degeneration of seminiferous tubules, and almost complete reduction of sperm count in the epididymal duct. Conclusion: Chronic hyperglycemia is therefore deleterious to the male reproductive system of mice. It leads to hypogonadism, which causes dysfunction of the male reproductive system, and can lead to infertility in men with diabetes mellitus.

Morphological and morphometric changes and epithelial apoptosis are induced in the rat epididymis by long-term letrozole treatment.

The epididymis is an organ that plays a key role in sperm maturation. The aim of this study was to examine the association between the chronic treatment of mature male rats with letrozole and morphological evaluation and morphometric values of epididymis as well as changes in the number of apoptotic cells in epididymal epithelium. Adult rats were treated with letrozole for 6 months and the epididymis weight, morphology, morphometric values and the number of apoptotic cells in the epithelium were examined. Long-term aromatase inhibition resulted in presence of intraepithelial clear vacuoles, hyperplasia of clear cells and a hyperplastic alteration in the epithelium known as a cribriform change. Moreover, changes in diameters of the epididymal duct and the epididymal lumen and changes in the epididymal epithelium height were observed. The number of apoptotic epithelial cells was increased in letrozole-treated group. It can be indicated that chronic treatment with letrozole can affect morphology, morphometric values and apoptosis in the epididymis of adult male rats. Observed changes are similar to that observed in the aging processes and may also be important for patients treated with aromatase inhibitors.

Three-dimensional analysis of mesonephric tubules remodeling into efferent tubules in the male mouse embryo.

Spermatozoa are transported to the epididymal duct through efferent tubules. Although the origin of the efferent tubules is thought to be mesonephric tubules (MTs), their detailed developmental process, for example, where the rete testis and efferent tubules are connected, is unclear. We investigated the structural changes of the MTs in the male mouse embryo using a three-dimensional reconstruction method. Three to six MTs were connected to the Wolffian duct, and some of them branched, resulting in five to nine tips. Rete cells contacted the three to six tips. The MTs showed a folded shape when the gonadal fate was determined. After the testis development started, they became short and straight but emerged as long and twisting by birth. Before birth, the efferent duct was composed of MTs and a cranial portion of the folded Wolffian duct. The contact between the rete testis and efferent tubules is possibly established at the tip of each MT. The MTs regress after gonadal fate is determined but is remodeled to the twisting efferent tubules by birth. The efferent tubules are composed of the MTs but also a cranial portion of the folded Wolffian duct in the mouse.

Effects of dipyrone and acetylsalicylic acid on contractions of distal cauda epididymis duct, serum testosterone and sperm count in rats.

The effects of dipyrone and acetylsalicylic acid (ASA) on male fertility are still not fully understood, mainly considering the epididymis as a putative target for their anti-fertility effects. Therefore, this study aimed to investigate the effects of dipyrone and ASA on the contractions of distal cauda epididymis duct, serum testosterone levels and sperm parameters in rats. Firstly, we checked the in vitro effects of dipyrone and ASA (10-1000 μM) on the contractions of distal cauda epididymis duct by pharmacological experiments. We also evaluated the effects of in vivo treatment with dipyrone and ASA 100 mg/kg (p.o.) for 15 days on epididymal duct contractions, serum testosterone levels and sperm parameters. In vitro dipyrone or ASA decreased the epididymal duct contractions induced by phenylephrine or carbachol. We observed that in vivo treatment with both drugs decreased the daily sperm production, serum testosterone levels and sperm count through epididymis without altering the epididymal duct contractions and sperm transit time through epididymis. In conclusion, in vitro dipyrone and ASA were able to diminish the contractions of epididymal duct, whilst in vivo administration decreased the sperm count throughout epididymis as a consequence of a low sperm production caused by reduced testosterone levels.

Histological, Histochemical, and Ultrastructural Approach to the Ductus Deferens in Male Nile Monitor Lizard (Varanus niloticus).

The ductus deferens is a fundamental part of the male genital tract and the continuation of the epididymal duct. As a male secondary sex organ, the ductus deferens plays a crucial role in the nourishment, storage, and maturation of spermatozoa. Some studies have provided information about the ductus deferens structure in reptiles; however, the full description of the ductus deferens remains to be clarified. The current study aimed to describe the Nile monitor lizard (Varanus niloticus) ductus deferens from histological, histochemical, and ultrastructural perspectives. The results revealed that the ductus deferens is formed histologically from two main cell types: principal and basal. The principal cells were tall and filled with periodic acid Schiff (+)/alcian blue (−) cytoplasmic granules. The basal cells were found just above the basement membrane. By transmission electron microscopy, the principal cells exhibited typical protein-secreting cell features. Additionally, some intraepithelial cells, such as halo cells, undifferentiated mesenchymal cells, and agranular leukocytes, were identified. This study presents the first detailed description of the Varanus niloticus ductus deferens. Further immunohistochemical studies are required to explore the function(s) of the cellular components.

Kcnj16 (Kir5.1) Gene Ablation Causes Subfertility and Increases the Prevalence of Morphologically Abnormal Spermatozoa.

The ability of spermatozoa to swim towards an oocyte and fertilize it depends on precise K+ permeability changes. Kir5.1 is an inwardly-rectifying potassium (Kir) channel with high sensitivity to intracellular H+ (pHi) and extracellular K+ concentration [K+]o, and hence provides a link between pHi and [K+]o changes and membrane potential. The intrinsic pHi sensitivity of Kir5.1 suggests a possible role for this channel in the pHi-dependent processes that take place during fertilization. However, despite the localization of Kir5.1 in murine spermatozoa, and its increased expression with age and sexual maturity, the role of the channel in sperm morphology, maturity, motility, and fertility is unknown. Here, we confirmed the presence of Kir5.1 in spermatozoa and showed strong expression of Kir4.1 channels in smooth muscle and epithelial cells lining the epididymal ducts. In contrast, Kir4.2 expression was not detected in testes. To examine the possible role of Kir5.1 in sperm physiology, we bred mice with a deletion of the Kcnj16 (Kir5.1) gene and observed that 20% of Kir5.1 knock-out male mice were infertile. Furthermore, 50% of knock-out mice older than 3 months were unable to breed. By contrast, 100% of wild-type (WT) mice were fertile. The genetic inactivation of Kcnj16 also resulted in smaller testes and a greater percentage of sperm with folded flagellum compared to WT littermates. Nevertheless, the abnormal sperm from mutant animals displayed increased progressive motility. Thus, ablation of the Kcnj16 gene identifies Kir5.1 channel as an important element contributing to testis development, sperm flagellar morphology, motility, and fertility. These findings are potentially relevant to the understanding of the complex pHi- and [K+]o-dependent interplay between different sperm ion channels, and provide insight into their role in fertilization and infertility.

Dose-related morphological changes in the epididymal region of sexually active adult male Japanese quail treated with di-n-butyl phthalate (DBP) commencing during the pre-pubertal stage

Di-n-butyl phthalate (DBP) is widely used as a plasticizer in personal care and medical products and is known to induce toxicity in the male reproductive organs in both mammals and birds. In this study, there was investigation of the effects of DBP on the epithelium of the rete testis, proximal, and distal efferent ductules and epididymal duct of adult Japanese quail (Coturnix japonica) following treatment with varying doses during the pre-pubertal and peri-pubertal periods. Pre-pubertal quail (n = 25) 4 weeks post-hatching were dosed orally with 10, 50, 200 and 400 mg DBP/kg/d, for 30 days and control birds were administered corn-oil only (n = 5 per group). Histo-metrically, there was lesser (P < 0.001) epithelial heights of the rete testis and efferent ductules in all quail DBP-treated groups, but not in the epididymal duct epithelium. There were no morphological change effects as a result of DBP treatments in the rete testis epithelium, while there were epithelial cytoplasmic vacuoles detected in the distal efferent ductule and epididymal duct of birds treated with 50, 200 and 400 mg DPB/kg/d. There were several lesions, including degenerative changes, cytoplasmic vacuoles, apoptosis and autophagy in the epithelium of the proximal efferent ductule in quail treated with 200 and 400 mg DBP/kg/d. Overall, the results indicate that treatment with DBP during the pre-pubertal period induced dose-dependent histometric and morphological changes in the epithelium of the epididymal region. It is concluded that the proximal efferent ductule was a highly sensitive component of the epididymal tissues of Japanese quail following treatment with DBP during the pre-pubertal period.

Paratesticular Papillary Cystadenoma of the Epididymis in the Setting of von Hippel-Lindau

An individual with von Hippel-Lindau disease presented with bilateral paratesticular masses. Prior germline testing had revealed the presence of a pathogenic VHL c.620T>C alteration. His past medical history was significant for right cerebellar hemangioblastoma, laser photocoagulation procedures for bilateral retinal hemangioblastomas, as well as multiple partial nephrectomies for clear cell renal cell carcinoma. In addition, he had also undergone a distal pancreatectomy for a well-differentiated neuroendocrine tumor. He was evaluated for worsening bilateral scrotal pain and a bilateral epididymectomy was performed for a 3.0-cm mass on the left and a 3.6-cm mass on the right. Representative images of the tumor arising within the epididymis (A) have been depicted. Specifically, this tumor exhibited papillary architecture (B) and was comprised of cells with clear cytoplasm that exhibited reverse polarity (ie, oriented away from the basement membrane; C). Immunohistochemistry for carbonic anhydrase IX, which is upregulated secondary to hypoxia-dependent signaling in tumors that have loss of function alterations of VHL, demonstrated a constitutive pattern of expression characterized by diffuse membranous localization (D). What is the most likely diagnosis?a. Metastatic clear cell renal cell carcinoma involving the epididymisb. Primary papillary cystadenoma of the epididymisc. Metastatic clear cell (tubulo-) papillary renal cell carcinoma involving the epididymisd. Primary adenocarcinoma of the epididymis Answer: b. Primary papillary cystadenoma of the epididymis. Papillary cystadenomas are benign tumors and are thought to arise from epididymal ducts.1Odrzywolski K.J. Mukhopadhyay S. Papillary cystadenoma of the epididymis.Arch Pathol Lab Med. 2010; 134: 630-633Crossref PubMed Google Scholar The mean age at presentation is approximately 35 years, and these tumors range in size from 0.5 to 8.0 cm.1Odrzywolski K.J. Mukhopadhyay S. Papillary cystadenoma of the epididymis.Arch Pathol Lab Med. 2010; 134: 630-633Crossref PubMed Google Scholar Although solitary tumors are identified more frequently, the presence of bilateral tumors should raise the possibility of von Hippel-Lindau disease.1Odrzywolski K.J. Mukhopadhyay S. Papillary cystadenoma of the epididymis.Arch Pathol Lab Med. 2010; 134: 630-633Crossref PubMed Google Scholar These tumors have significant morphologic and immunophenotypic overlap with both clear cell renal cell carcinoma as well as clear cell (tubulo-) papillary renal cell carcinoma.2Cox R. Vang R. Epstein J.I. Papillary cystadenoma of the epididymis and broad ligament: morphologic and immunohistochemical overlap with clear cell papillary renal cell carcinoma.Am J Surg Pathol. 2014; 38: 713-718Crossref PubMed Scopus (20) Google Scholar For instance, neither the expression of PAX8 or diffuse membranous localization of carbonic anhydrase IX (a surrogate marker of hypoxia-dependent signaling in tumors that have loss of function alterations of VHL) can be used to reliably distinguish clear cell type renal cell carcinoma from primary papillary cystadenoma of the epididymis.2Cox R. Vang R. Epstein J.I. Papillary cystadenoma of the epididymis and broad ligament: morphologic and immunohistochemical overlap with clear cell papillary renal cell carcinoma.Am J Surg Pathol. 2014; 38: 713-718Crossref PubMed Scopus (20) Google Scholar The diagnosis of papillary cystadenoma of the epididymis is therefore established based on a combination of histopathologic examination as well as the clinical context, including tumor site.

In Vitro Contraction of Isolated Cauda Epididymal Duct Smooth Muscle as a Complimentary Approach to Physiological, Pathological, Toxicological, and Pharmacological Studies on Epididymal Function.

Contraction of cauda epididymal duct (CE) smooth muscle is one of the very first events of the seminal emission phase of ejaculation. The contraction of CE smooth muscle is governed by a complex interaction of hormones, autacoids, and by the neurotransmitters released from the epididymal intramural nerve endings, and any impairment in the CE smooth muscle contraction has the potential to impair male fertility. Apart the obvious pathophysiological and toxicological importance of CE smooth muscle contraction, modulation of CE contraction has pharmaceutical interest offering a druggable target to development of drugs to improve/impair male fertility. The in vitro contraction experiments constitute a valuable approach to an in-depth evaluation of functional and molecular changes resulting from pathologies or drug exposure. Therefore, this chapter consists in a description of in vitro pharmacological reactivity contractility of the epididymal duct in a controlled medium, maintained at 30°C of temperature and continuously bubbled with 95% O2 and 5% CO2 to obtain cumulative concentration-response curves that has been fundamental to some of our investigations on epididymal physiology, toxicology, and pharmacology.

Immunoreactivity Of Alpha Smooth Muscle Actin In The Epididymis Of The Dromedary Camel: Impact Of The Sexual Maturity And The Breeding Seasonality

This study was carried out to highlight the seasonal variation of the immunoreactivity of the protein alpha smooth muscle actin (a-SMA), within the epididymal duct of the male dromedaries. Immunostaining for detection of a-SMA was applied on paraffin-embedded sections taken from different regions of the epididymal duct from both juvenile and adult dromedaries, during the different seasons of the year. Immunoreactivity (IR) was mainly confined to the smooth muscle cells (SMCs) within the peritubular muscle coat and in the walls of the blood vessels. The intensity of IR displayed a remarkable seasonal variation in adults. The strongest IR has been recorded in all epididymal regions (head, body and tail), during the breeding season (corresponds to winter months). In the season of breeding inactivity (corresponds to summer months), the a-SMA-IR exhibited the lowest intensity. During the period of transition from activity to inactivity (corresponds to spring months) and from inactivity to activity (corresponds to autumn months), a moderate a-SMA-IR has been reported. On the other hand, a weak to moderate immunoreactivity for a-SMA appeared mainly in innermost layers of the peritubular cells surrounding the epididymal duct of the juvenile dromedaries throughout the year. In conclusion, both of the sexual maturity and the breeding seasonality demonstrated a clear impact on the immunostaining for a-SMA in male dromedaries. This may be relevant to the physiological alterations that are linked to hormonal control throughout the year.

Research on the protective effect of caffeic acid phenethyl ester on testicular damage caused by cisplatin

Background/aimCisplatin (CP), a chemotherapeutic drug, causes damage to spermatogenic serial cells, Sertoli cells, and Leydig cells in rat testicles. It was aimed to investigate the protective effect of caffeic acid phenethyl ester (CAPE), one of the active ingredients of propolis, in eliminating CP-induced testicular damage.Materials and methodsGroup 1 (control group) was given physiological saline solution intraperitoneally (IP) throughout the experiment. Group 2 (CP group) was given a single dose of CP (7 mg/kg) IP on the day 7. Group 3 (CP + CAPE group), was given CAPE (10 µmol/kg/day) IP for 12 days and a single dose of CP (7 mg/kg) IP on day 7. Group 4 (CAPE group) was given CAPE (10 µmol/kg/day) IP for 12 days. On day 14 of the experiment, the rats were decapitated under xylazine and ketamine anesthesia and their testicles were removed. The sections obtained from the testicles were stained with hematoxylin-eosin and histopathological damage was evaluated. Malondialdehyde (MDA) levels, and superoxide dismutase (SOD) and catalase (CAT) enzymatic activities were measured in the testicular tissue samples. Testosterone (TES) levels were measured in the blood serum. The Johnsen testicular biopsy score (JTBS) was used to evaluate testicular tubules. DNA damage was evaluated in sperm samples taken from the ductus epididymis using the comet assay technique.ResultsIn Group 2, which was given CP, the testicles were severely damaged. It was observed that histological damage was reduced in the testes by administering CAPE in Group 3. Moreover, according to the JTBS, the value was significantly higher in the testicular tubules (P < 0.05). Moreover, the MDA level decreased in Group 3. However, the SOD, CAT, and TES levels increased in Group 3. DNA damage also decreased significantly in Group 3 when compared to Group 2 (P < 0.05).ConclusionThe results showed that CAPE may be protective against damage caused by CP in the testicles of rats.

Impact of diabetic hyperglycaemia and insulin therapy on autophagy and impairment in rat epididymis

Blood glucose dysregulation and hyperglycaemia caused by diabetes mellitus are intimately associated with male infertility. Two-month-old Sprague-Dawley rats were given a single dose of streptozotocin (55mg/kg) by intraperitoneal injection to induce type I diabetes mellitus (DM group). The treatment group was given 1 unit/day of insulin for 16weeks (INS group). The normal control group (NC group) was given food ad libitum. In the DM group, the histological analysis of caput and cauda epididymal ducts showed broken stereocilia and more lipid vacuolisations in the principal cells. The interstitial hyperplasia and inflammatory cell infiltration were observed in epididymal tissues. Transmission electron microscopy observation showed that the principal cells in the DM group contained more vacuoles, partly lost stereocilia, and swollen mitochondria. The autophagosomes were observed as well. Western blotting results of LC3II/I and P62 protein expression indicated that autophagy was downregulated in the DM group. The total antioxidant activity and GPx5 expression of epididymal tissues were also decreased. In the INS group, significant improvements were observed in epididymal tissues. Our study suggests that diabetic hyperglycaemia causes autophagy dysregulation in epididymal tissues, which may play a role in diabetes-induced rat epididymal injury. Insulin treatment is beneficial for diabetic-associated epididymal dysfunction.

Mouse defensin beta 20 (Defb20) is expressed specifically in the caput region of the epididymis and regulated by androgen and testicular factors

Sperm cells undergo maturation during their transit throughout the epididymis. This process takes place in region-specific manner in which sperm are battered by proteins secreted by epithelium lining the epididymal duct. Most of the genes that encode for the proteins involved in the sperm maturation remain uncharacterized. Previous studies showed that family of β-defensins preferentially eaxpressed in male reproductive tracts and play an important role in both innate immunity and sperm fertility. In this study we characterized Defb20 to gain insight on its role in sperm maturation. Bioinformatic tools were used to analyzed functional domains and signal peptide. qRT-PCR analyses were used to analyzed tissue distribution, dependency on androgen and testicular factors and developmental-regulated expression analysis. Defb20 sequence contains important domains such as N-myristoilation and kinase binding sites which are putatively involved in the protein activation and protein-plasma membrane interaction. Moreover, DEFB20 contains a signal peptide indicating characteristic of secretory proteins. Defb20 was expressed exclusively in the epididymis with the highest expression in the caput region and was down-regulated by gonadectomy. Defb20 was also regulated by testicular factors in which the expression was down-regulated after efferent duct ligation (EDL). The dependency on the androgen was further confirmed by postnatal expression analysis in which Defb20 began to express at day-20 postnatal indicating specific stage of expression after initial development of the testis. In conclusion, Defb20 have a potential to be involved in the epididymal sperm maturation process.

Immunolocalization of androgen and vitamin D receptors in the epididymis of mature ram (Ovis aries).

This study illustrated the immunohistochemical distribution of androgen and vitamin D receptors of epididymis in 20 sexually mature ram (Rahmani breed) with average age ranged from (2_4) years and average weight ranged from (50_65kg). Androgen receptor was localized in the cytoplasm of both ciliated and non ciliated cells of efferent ductules, besides the principal cells via the entire epididymal duct. The principal cells of both corpus and proximal cauda epididymis showed the highest immunoreactivity to androgen receptors. Furthermore, vitamin D receptor was localized in the cytoplasm of all epithelium of the efferent ductules besides principal cells of all epididymal regions, however the immunoreaction was significantly higher in the efferent ductules, distal caput and distal cauda epididymis. In conclusion, these results suggest that the function of ram epididymis is regulated by both androgen and Vitamin D.

The Involvement of the Chemokine RANTES in Regulating Luminal Acidification in Rat Epididymis.

BackgroundA complex interplay between different cell types in the epithelium leads to activation of the luminal acidifying capacity of the epididymis, a process that is crucial for sperm maturation and storage. Basal cells sense the luminal angiotensin II (ANG II) and stimulate proton secretion in clear cells through nitric oxide (NO). Our previous study has shown the chemokine regulated upon activation normal T-cell expressed and secreted (RANTES) was expressed in the F4/80 positive macrophages of human epididymis. The objective of this study was to explore the involvement of RANTES in regulating the luminal acidification in the rat epididymis.MethodsThe role of RANTES was investigated by in vivo perfusion with recombinant RANTES, Met-RANTES, and PBS of different pH values. Furthermore, rats vasectomy was performed to alter the epididymal luminal pH. RIA was used to measure the tissue homogenate ANG II concentration. Real time-PCR and western blot were employed to examine the expression levels of AGTR2, RANTES, CCR1, CCR5, and iNOS in epididymis.ResultsRANTES was restricted to the basal macrophages of epididymal ducts and co-localized with its receptors CCR1 and CCR5. Both V-ATPase and iNOS were up-regulated in the cauda epididymis after perfused with recombinant RANTES, while the antagonist Met-RANTES perfusion led to a complete abrogation of the increased expression of V-ATPase in the apical membrane of clear cells and iNOS in macrophages. Upon alkaline perfusion, RANTES expression was significantly increased and the apical accumulation of V-ATPase in the clear cells was induced in the cauda epididymis. The luminal pH in the cauda epididymis increased after vasectomy. The concentration of the ANG II and the expression levels of AGTR2, RANTES, CCR1, CCR5, and iNOS dropped in the cauda epididymis following vasectomy.ConclusionUpon the activation of basal cells, RANTES might induce the NO release from macrophages by interacting with its receptors, which increases proton secretion by adjacent clear cells. Thus, RANTES is possible to participate in the crosstalk among basal cells, macrophages and clear cells for the fine control of an optimum acidic luminal environment that is critical for male fertility.

Postnatal differentiation and regional histological variations in the ductus epididymidis of the Congjiang Xiang pig

The Congjiang Xiang pig is a rare Chinese miniature breed whose epididymal histologic features through the postnatal development are poorly understood. To clarify the histomorphological differences between each region of epididymis during postnatal development, 24 male Congjiang Xiang pigs aged from neonatal (15 d), peri-puberty (30 d), puberty (60 d) to adult (180 d) stages, were examined. Postnatal differentiation of the different regions (I–V) of the epididymis started from birth and continued until maturity that showed regional variations. Developmental progression was disto-proximal. At the neonatal stage, Wolffian duct differentiation starts in the distal region, then ascends to the middle region which forms regions V, IV and III, respectively. A simple lined cuboidal in the epidydimal epithelial, which gradually differentiated into a pseudostratified columnar with stereocilia from neonatal to post-pubertal. After puberty cell rearrangement occurred in the epithelium, differentiation accelerated, and spermatozoon seen in the lumen, especially the lumen of region II. In region III, both halo and apical cells were frequently observed. At the post-pubertal stage, clear cells were frequently observed in Region IV-V, and the epididymal duct was markedly increased in size and fully packed with spermatozoa. The information presented in this study will be helpful for future evaluations of Congjiang Xiang pig fertility. After puberty cell rearrangement occurred in the epithelium, differentiation accelerated, and spermatozoon seen in the lumen, especially the lumen of region II. In region III, both halo and apical cells were frequently observed. At the post-pubertal stage, clear cells were frequently observed in Region IV–V, and the epididymal duct was markedly increased in size and fully packed with spermatozoa. The information presented in this study will be helpful for future evaluations of Congjiang Xiang pig fertility.

Contractions transport exfoliated epithelial cells through the neonatal epididymis.

Contractions of the adult epididymal duct are well known in the context of sperm transport. Some reports also describe contractions of the epididymal duct during development, but data about their character, regulation and function are sparse. In the foetal human epididymis we found luminal cells and could identify them as exfoliated epithelial cells originating from the epididymis and not from testis by using antibodies against neutral endopeptidase as an epithelial epididymal duct marker. Exfoliated cells were also found in the epididymal duct after birth. Time-lapse imaging revealed directional transport of luminal cells in the neonatal rat epididymis interrupted by pendular movement. Spontaneous contractions were discovered in the neonatal epididymis and an association between these contractions and the transport of the luminal cells could be observed. Both, transport and spontaneous contractions, were affected significantly by substances known to contract (noradrenaline) or relax (the phosphodiesterase 5 inhibitor sildenafil) smooth muscle cells. Immunohistochemistry showed staining for the proliferation marker proliferating-cell-nuclear-antigen (PCNA) in cells of the ductal lumen of the neonatal rat epididymis indicating the extrusion of cells also during proliferation. Our data showed spontaneous contractions of the immature epididymal duct associated with the transport of exfoliated luminal cells before the first occurrence of sperm cells. Results suggest an important role including both (i) a mechanical place holder function of exfoliated luminal cells (ii) together with a novel idea of organized waste disposal of these cells during development.

Impact of thermal stress exposure on seminal quality, antioxidant defence system, TNF-α and TIMP-3 in Ossimi ram.

Heat stress (HS) occupies huge importance nowadays as it leads to severe economic losses especially in livestock. Preserving sheep against HS is one of the governmental scopes where it represents huge percentage of global ruminant. The present research was conducted to study semen quality, some stress and inflammatory markers in Ossimi rams under both hot and mild climatic conditions. The current study was conducted on selected 46 ram samples divided into two groups during summer and winter. Semen analysis, testosterone (TES), cortisol (COR) and blood glucose (BG) levels, and lipid and protein profiles were done. Concentrations of tumour necrosis factor alpha (TNF-α), tissue inhibitor of metalloproteinase-3 (TIMP-3), nitric oxide (NO), malondialdehyde (MDA) and reduced glutathione (GSH) and specific activity of glutathione peroxidase (GPx) and superoxide dismutase (SOD) were assessed. The results in summer compared to winter revealed significant elevation of total defects and number of dead sperms; however, there was reduction in sperm total motility and concentration and external epididymal tail duct diameter. Histological study of epididymal tail lumen exhibited azoospermia. Further, TES, TIMP-3 and GSH levels were decreased and COR, TNF-α, NO and MDA were raised. Specific activities of GPx and SOD were also declined. Additionally, there was a significant increase in concentrations of BG and lipid profiles except high-density lipoprotein. Our data concluded that there were new insights into TNF-α and TIMP-3 as biomarkers can be used in diagnosis of sheep suffering from HS, but further studies are recommended to do in future work about such aspect.