Histochemical changes in the testis and epididymis after vasectomy

Bilateral vasectomy was performed in 10 adult dogs, using another set of 10 as a matching control. Both normal testes were removed from 1 vasectomized and 1 normal dog at weekly intervals. The distribution of alkaline phosphatase, glycogen, lipids, and cholesterol was studied histochemically. The epididymis was also studied. Marked degeneration occurred in the seminiferous tubules, and spermatogenesis ceased. Alkaline phosphatase and lipids decreased appreciably, but the basal cells (spermatogonia) remained practically unaffected. There was thickening of the PAS positive, diastase resistant tunica propria from the fourth week on. The lydig cell clusters became more prominent and showed increase in cholesterol and lipids. The epididymis appeared distended but without any visible sperms. The epithelial cells were markedly flattened. Alkaline phosphatase decreased near the lumal border, but the nuclei, now packed at the periphery, gave a more intense reaction. The functional significance of these changes, particularly in view of the large scale use of vasectomy today, is discussed.     

Qualitative and quantitative changes of acid and alkaline phosphatases in the testis and epididymis of mice in relation to single high dose of alpha-chlorohydrin

The present study is designed to know whether the enzymes related to permeability and general metabolism of the cells of testis and epididymis are affected by alpha-chlorohydrin. For this purpose two enzymes, viz., acid and alkaline phosphatases are studied thoroughly by qualitative and quantitative parameters during single high dose (90 mg/kg body weight) treatment after 24 and 48 hours of drug administration. Along with this, the changes in the behaviour of the animals and histological structure of testis and epididymis of mice are also recorded.     

Successful pregnancies after using immotile spermatozoa from ejaculate,epididymis and testis

OBJECTIVE: To assess the efficacy of intracytoplasmic sperm injection (ICSI) in term of pregnancy rate with immotile spermatozoa from ejaculate, epididymis and testis. STUDY DESIGN: A retrospective study was conducted between January 1998 and March 2001. We performed intracytoplasmic sperm injection with immotile spermatozoa, in 160 couples during 172 cycles. RESULTS: The birth rate per cycle was 38.4% in immotile spermatozoa from ejaculate, 35.4% from testis and 38.7% from epididymis. CONCLUSION: This retrospective analysis shows that immotile spermatozoa retrieved from epididymis or testicle gives similar fertilization and pregnancies rates as immotile spermatozoa from ejaculate.     

Oxidative stress,spermatozoa and leukocytic infiltration: relationships forged by the opposing forces of microbial invasion and the search for perfection

This review addresses the complex relationships that exist between spermatozoa and the immune system and highlights the role of oxidative stress in regulating the direction and functional relevance of these interactions. Spermatozoa are potentially antigenic; however, in the testes and epididymis these cells are sequestered behind physical barriers and benefit from a tolerogenic state generated through the mediation of indoleamine dioxygenase. In the female there are no such barriers; however, inseminated spermatozoa are protected by the concomitant presence of seminal plasma. The latter possesses immunosuppressive properties, a powerful array of antioxidants and cytokines that modulate the immunological response to semen deposition. Subsequent to insemination, leukocytic infiltration of the female tract occurs to facilitate the removal of millions of residual moribund and senescent spermatozoa, while allowing the most competent cells to ascend to the site of fertilization. The post-insemination phagocytosis of non-viable spermatozoa is ‘silent’ in the sense that no reactive oxygen species (ROS) or pro-inflammatory cytokines are generated. The silent phagocytosis of senescent spermatozoa is a response to markers, such as phosphatidylserine, which are expressed on the surface of spermatozoa as they engage in the intrinsic apoptotic cascade. By contrast, infection can bring fully activated leukocytes into the male reproductive tract that are actively generating ROS and releasing pro-inflammatory cytokines. Such free-radical-generating leukocytes have the potential to seriously damage the functionality of spermatozoa as well as the integrity of their DNA, particularly in vitro, when these cells are devoid of the antioxidant protection afforded by seminal plasma.     

Micropuncture and microanalytic studies of the effect of vasectomy on the rat testis and epididymis.

Micropuncture techniques adapted for in vivo use in the male reproductive tract were used to study the effect of vasectomy on the concentration and morphology of spermatozoa in the seminiferous tubules and ductus epididymidis of the rat. Regardless of surgical technique, 85% of all rats which had previously under vasectomy developed a speratic granuloma at the proximal end of the ligated vas deferens. Vasectomy did not affect the weights of the ipsilateral (vasectomized) testes. Vasectomy without granuloma formation resulted in an elevation in the weight of the ipsilateral epididymis and an increase in the concentration of spermatozoa in the ipsilateral cauda. Vasectomy complicated by granuloma formation caused a significant increase in the ratio of the spermatocrit in the seminiferous tubule to that in the caput. In the vasectomized rat without granuloma, significantly more abnormal spermatozoa were in the caput than in the cauda; in the normal rat, the concentrations of abnormal spermatozoa in the two regions were not significantly different. Vasectomies with and without granuloma formation are distinct phenomena and should be investigated separately.     

Interaction between Smad1 and p97/VCP in rat testis and epididymis during the postnatal development

Members of the bone morphogenetic proteins (BMPs) superfamily are expressed in the testis and epididymis and are believed to have different biological functions during testicular and epididymal development. Smad1 is one of the signal transducers of BMP signaling and binds to several proteins involved in ubiquitin-proteasome system (UPS). Valosin-containing protein (p97/VCP) is required for the degradation of some UPS substrates. Although p97/VCP has been indicated in different cellular pathways, its association with BMP signaling in male reproductive system has not been elucidated. The aim of the present study was to investigate the cellular localization of Smad1, phospho-Smad1, and p97/VCP and the interaction of proteins in the postnatal rat testis and epididymis. Testicular and epididymal tissues from 5-, 15- and 60-day-old rats were examined by immunohistochemistry, immunofluorescence, Western blotting, and immunoprecipitation techniques. In 5-day-old rat testis, Smad1, phospho-Smad1, and p97/VCP were mainly expressed in gonocytes. In 15- and 60-day-old rat testis, proteins were overlapped in spermatogonia, Sertoli cells, and spermatocytes. Expression of proteins in the epithelial cells of epididymis was gradually increased from 5 to 15 days of age. Smad1 and phospho-Smad1 expressions showed uniformity in the different regions of epididymis, however p97/VCP immunoreactivity was higher only in caput epididymis compared to corpus and cauda epididymis in 15- and 60-day-old rat epididymis. Co-immunoprecipitation experiments further confirmed the Smad1-p97/VCP and p-Smad1-p97/VCP interactions. The overlap between Smad1 and p97/VCP expressions in the postnatal rat testis and epididymis suggests that p97/VCP may play important roles in mediating BMP signaling during spermatogenesis.     

Electrophoretic and immunocytochemical analysis of Hsp72 and Hsp73 expression in heat-stressed mouse testis and epididymis

Objective

One of the most profound events in stressed cells is the synthesis of a highly conserved family of proteins, the ‘heat shock proteins’ (Hsp). The Hsp70 family is the most diverse, and includes constitutive as well as stress-inducible proteins with overlapping or unique functions in different cell compartments. Elucidation of Hsp70 expression during different stages of spermatogenesis and maturation of germ cells is of particular interest due to their high sensitivity to heat treatment.

Study design

Expression of the main isoforms of the Hsp70 family (constitutive Hsp73 and stress-inducible Hsp72) was determined in normal and heat-stressed mouse testes and epididymis from sexually mature (60-day-old) mice during spermatogenesis and maturation of germ cells. Immunocytochemical analysis and one- and two-dimensional gel electrophoresis were used to separate mouse testicular and epididymal proteins from saline extracts, followed by Western blotting.

Results

Using a polyclonal anti-Hsp70 antibody that recognizes both isoforms, inducible Hsp72 expression, was demonstrated immunocytochemically only in heat-stressed tissues, while a high level of constitutive Hsp73 isoform expression was found in both normal and heat-stressed mouse male reproductive tissues. Morphological studies have shown that round and elongated spermatids in the testes, as well as all segments of the epididymis, are most sensitive to heat stress. In the epididymis, the reaction was localized in different cell compartments.

Conclusion

In heat stress conditions, Hsp73 is mobilized to prevent apoptosis in the testes and epididymis, and assists Hsp72 in the repair of stress-altered protein conformations.     

Tissue immunoexpression and messenger ribonucleic acid localization of inhibin/activin subunit in human epididymis

OBJECTIVE: To determine the expression of inhibin betaA and betaB subunits and follistatin and the ability of human epididymal epithelium to synthesize these molecules. DESIGN: The main aim of this study was to investigate the expression of inhibin alpha, betaA, and betaB-subunits and follistatin in human epididymis with immunohistochemistry, in situ hybridization, and Western blotting in adult life. SETTING: Academic university hospital. PATIENT(S): Epididymes were obtained from 10 men undergoing routine vasectomy or surgery for benign disease at the Royal Hallamshire Hospital, Sheffield, United Kingdom. MAIN OUTCOME MEASURE(S): Immunoexpression of activin betaA and betaB subunits and follistatin proteins and mRNA in human caput and cauda epididymis. RESULT(S): Positive immunoexpression for activin betaA and betaB subunits and follistatin were detected in different parts of the epididymis epithelium. Western blotting under a reducing condition detected a 28-kd band (possibly corresponding to the activin dimer). In situ hybridization indicated positive mRNA localization signal in both caput and cauda epididymal epithelium. CONCLUSION(S): Activins betaA and betaB subunits, but not inhibin alpha subunit, were detected in epididymal epithelium. These finding suggest that activins might have a role in the processes of sperm maturation and sperm fertilizing capability during transit and storage.     

Tissue testosterone and dihydrotestosterone from bilateral testis biopsies in males with varicocele.

Nine males with varying degrees of subfertility with left varicocele underwent bilateral testicular biopsy for determination of testicular testosterone (T) and dihydrotestosterone (DHT) content. Testicular tissue T and DHT levels varied greatly, and there was no significant difference when the group T and DHT content of the left testis was compared with that of the right testis. When the respective mean preoperative sperm count was compared with the bilateral tissue T and DHT content, no consistent relationship was found.     

Biochemical observations on the protein and nucleic acid metabolism of the rat testis and epididymis after treatment with low doses of alpha-chlorohydrin

A biochemical study was made on the effects of low doses of alpha-chlorohydrin on the protein and nucleic acid metabolism of the rat testis and epididymis. RNA and protein levels were decreased both in the testis and epididymis. The DNA content of the testis and epididymis did not change after exposure of the animals to the drug. The reduced concentrations of RNA and protein were closely paralleled by the increased activity of proteinase (protein hydrolyzing enzyme) and ribonuclease (RNA degrading enzyme) in the testis and epididymis. The gamma-glutamyl transpeptidase activity of both the testis and epididymis was also reduced indicating the slow transfer of amino acids across the cell membranes of testis and epididymis and thus low protein synthesis. The DNAase levels of rat testis and epididymis did not show any appreciable change in response to the alpha-chlorohydrin treatment. These studies indicate that although there may not be any histological damage in the tissue the metabolic pathways may become defective much earlier before any visible morphological change is discernible.     

Effect of curcumin on aflatoxin-induced biochemical changes in testis of mice

OBJECTIVE: To investigate the effect of curcumin on aflatoxin-induced biochemical changes in testis of mice. DESIGN: Controlled research laboratory study. SETTING: Aflatoxin was obtained by growing Aspergillus parasiticus in SMKY liquid medium. Pure curcumin (97% purity) was purchased from Hi-Media Laboratories Pvt. Ltd., Mumbai, India. PATIENT(S): Young inbred, Swiss strain male albino mice (Mus musculus), weighing approximately 37-40 g, were obtained from Cadila Health Care, Ahmedabad, India. INTERVENTION(S): Aflatoxin was administered orally at 25 microg (low dose) and 50 microg (high dose)/0.2 mL olive oil/animal/day (750 and 1,500 microg/kg body weight), respectively, with and without curcumin for 45 days. On the 46th day the animals were sacrificed by cervical dislocation. Testis were removed and weighed. Homogenates were prepared for respective parameters such as protein, cholesterol, DNA, RNA, and 3beta- and 17beta-hydroxysteroid dehydrogenase activity. MAIN OUTCOME MEASURE(S): Protein, cholesterol, DNA, RNA, 3beta- and 17beta-hydroxysteroid dehydrogenase activity were measured immediately. RESULT(S): There was a dose-dependent effect of aflatoxin on testis of mice. Protein, cholesterol, DNA, RNA, and 3beta- and 17beta-hydroxysteroid dehydrogenase activity were reduced significantly by the treatment of aflatoxin; cholesterol was increased. Treatment with curcumin along with aflatoxin ameliorates aflatoxin-induced biochemical changes in the testis of mice. CONCLUSION(S): Curcumin significantly ameliorates aflatoxin-induced biochemical changes in the testis of mice.     

Correlative histochemical and biochemical studies on the adenosine triphosphatase, succinic dehydrogenase and acetylcholinesterase in the epididymis of mice after alpha-chlorohydrin treatment

The present paper deals with the correlative histochemical and biochemical studies of the epididymis following the treatments of alpha-chlorohydrin. This drug was administered in chronic low dose (15 mg/kg body weight/day) for 20 and 30 days and a single high dose (90 mg/kg body weight). Histochemical alterations of ATPase, SDH and AChE were studied in various components of epididymal epithelium and the total enzyme content was measured by biochemical parameters. The study shows progressive decrease of the enzymes in the interstitium and the epithelium of both the caput and cauda epididymes with increasing dose and duration, except for the high dose effect of alpha-chlorohydrin on AChE. Since alpha-chlorohydrin decreases the androgen dependent enzymes (ATPase, SDH, AChE), there is a possibility that the drug may be antiandrogenic in nature. In such case the action of these drugs may not be directly on the spermatozoa, as proposed by earlier workers, but is mediated by changing the physiology of the epididymis, affecting the milieu in which the spermatozoa mature.     

Structure of rete testis, vas efferens, epididymis and vas deferens of langur monkey (Presbytis entellus entellus Dufresne)

Morphological and histological features of rete testis, vas efferens, epididymis and vas deferens were studied in the langur monkey. Tubular extensions of rete were located towards lateral side of the testis. Its epithelium comprises mostly of cuboidal cells with hyaline cytoplasm. Three to nine bundles of vas efferens, emerging below the cranial pole of the testis, were observed. Vas efferens epithelium comprises of ciliated and nonciliated cells. Epididymis could be divided into six zones on the basis of cytological features. Principal cells, basal cells, apical cells and intraepithelial lymphocytes were observed in the epididymal epithelium, but their number, shape, size and location of nuclei varied in different zones. Vas deferens epithelium comprises of principal cells, basal cells, apical cells and few intraepithelial lymphocytes. Epithelium is surrounded by lamina propria, longitudinal, circular and longitudinal muscle layers.     

In defense of a function for the human epididymis

In view of reports that the human epididymis may play no role in human fertility, literature on the fertilizing capacity of epididymal spermatozoa was reviewed. The survey indicates that under the circumstances of their retrieval, human epididymal and testicular spermatozoa may have the ability to fertilize human eggs both in vivo and in vitro. Although the "fertility profile" of the normal epididymis cannot be explored in man, a fair assumption would be that fertilizing capacity develops fully in the distal part of the tract, judging from the higher motility and egg fusing ability of sperm taken from these regions of unobstructed tissue. Motility and fertilizing capacity observed with IVF or artificial insemination, in which sperm are obtained from obstructed ducts, may occur at a level in the tract more proximal than normal, as in animals. The pregnancies resulting from aspiration of spermatozoa from, or anastomosis of the vas deferens to, the efferent ducts are of great clinical interest, but the pathological state of the tissue precludes definitive statements about the functioning of a normal epididymis. In the former case, the immediate origin of the fertilizing spermatozoon and the nature of the secretions previously bathing it are unknown and in the latter case the time needed before pregnancies occur is much greater than anticipated had fertile sperm been present in the proximal epididymis. The evidence supports neither the view that testicular sperm are inherently fertile nor that a simple aging of sperm cells is sufficient for the fertilizing potential of spermatozoa to be realized. It emphasizes, rather, the importance of the environment to which the sperm cells are subjected. Under abnormal conditions other accessory glands may secrete compounds that are necessary for the maturation of spermatozoa. Before more information is known of the exact situation existing, or having existed, in pathological human tissues from which fertilizing human spermatozoa can be obtained, great caution should be exercised in interpreting the results of pregnancies arising from the in vivo and in vitro insemination of testicular or epididymal spermatozoa.