Effect of adrenalectomy on rat epididymidis

Aim:To investigate the effect of adrenalectomy (ADX) on the epididymidis of Sprague-Dawley rats.Methods:The histological,biochemical(cholesterol protein,zinc,copper,alkaline and acid phosphatase aryl sulphatase,lactic dehydrogenase and leucine amino peptidase)and hormonal (FSH,LH and testosterone) changes of caput and cauda epididymis in ADX rats were observed.Results:Organ wet weight,histological studies and morphometric measurements indicated a cellular degeneration in caput and cauda epididymis of ADX rats.Serum testosterone level was significantly lower in ADX than in sham-operated rats,while the serum FSH and LH were below the detection limit of 1 mIU/mL.The enzymatic activity was higher in ADX than in sham-operated rats.Epididymal zinc level increased whereas copper level decreased in ADX rats compared to the sham-operated.Conclusion:Adrenalectomy leads to degeneration of caput and cauda epididymidis epithelial cells as a result of decreased supply of testosterone.     

Effect of adrenalectomy and hydrocortisone on ventral prostate of rats

Aim: To study the effects of adrenalectomy and hydrocortisone on the ventral prostate of SD rats. Methods: Inadrenalectomised (ADX) and ADX hydrocortisone (1, 2, or 4 mg) treated rats, the prostatic histology and thecholesterol, protein, zinc, and copper levels and the enzymic profile (acid phosphatase, alkaline phosphatase, aryl sul-phatase, lactic dehydrogenase, and leucine aminopeptidase) in the prostatic tissue were determined; the serum hormon-al profile (testosterone, FSH and LH) was also assayed. Results; Adrenalectomy caused a progressive degenerationin prostatic structure that was not reversed by hydrocortisone treatment. The serum testosterone were significantly lowerin ADX than in sham operated rats and lower in ADX hydrocortisone than in ADX-C rats (P < 0.01). The serumFSH and LH were below the detection limit of 1 mIU/mL. The enzymatic activity was higher in ADX than in sham op-erated rats and higher in ADX hydrocortisone than in ADX-C rats (P<0.05-0.01). The prostatic zinc levels wer     

Androgens in various fluid compartments of the rat testis and epididymis after hypophysectomy and gonadotropin supplementation

Hypophysectomized male rats were administered LH or LH + FSH for 14 days and subjected to in vivo micropuncture collection of reproductive tract fluids to determine if FSH alters the compartmentalization of testosterone in the rat testis or of 5 alpha-dihydrotestosterone (DHT) in the epididymis. Testosterone and DHT concentrations were determined in cardiac blood serum, testicular venous serum, testicular interstitial fluid, and seminiferous tubule fluid, and in intraluminal fluid and tissue extracts from the caput and cauda epididymidis. Testosterone is the predominant androgen in the testis, and compared with control values, concentrations in venous sera, interstitial fluid, and tubule fluid were returned to values indistinguishable from controls by supplementation with 24 micrograms LH/day. 24 micrograms LH + 24 micrograms FSH/day did not augment the intratubular partition of testosterone. Epididymal DHT values were returned to control levels by LH alone, but additional supplementation with FSH significantly increased DHT from the caput epididymidis even further. It is speculated that FSH does not alter the compartmentalization of androgens in the rat testis, but may play a role in retaining androgens in the epididymis.     

In vitro Metabolism of (3H)-Androstenedione in the Rat Epididymis and Vas Deferens

The in vitro metabolism of (3H)-androstenedione in the epididymis and vas deferens of intact and castrated rats was investigated and the metabolites formed were identified by radio gas chromatography. Incubation of slices of caput epididymidis for 2 hr at 34 degrees C metabolised 90% androstenedione. Similar incubations of tissue samples from cauda epididymidis and vas deferens metabolized 60 and 25% of androstenedione respectively. The major metabolites formed in the epididymis were androstanedione (caput: 48%; cauda: 33%) and androsterone (caput: 35%; cauda: 13%). These metabolites appeared in much less concentration in the incubations with vas deferens (about 8% each). In general, conversion to testosterone and dihydrotesterone was low in all the three organs examined. Castration did not significantly alter the metabolic pattern in the caput epididymidis and vas deferens but promoted the formation of androsterone (38%) in the cauda epididymidis. The conversion of androstenedione, a weak androgen to testosterone, dihydrotestosterone and 3 alpha/3 beta-diols in the epididymidis and vas deferens of castrated rats may be of physiological significance. In addition, androsterone appears to be an important androgenic metabolite in the epididymis.     

Regional Differences in Steroidogenesis and Hormone Levels in the Epididymis and Vas Deferens of Adult Rats

In vivo and in vitro studies with different parts of the epididymis and vas deferens were carried out to determine their inherent capacity to synthesize steroids and to correlate with the endogenous levels with or without the administration of hCG.
Incubation with ¹⁴C-labelled pregnenolone and testosterone demonstrated that caput epididymidis was more active than other parts in synthesizing testosterone from ¹⁴C-pregnenolone and in converting labelled testosterone to 5α-dihydrotestosterone (DHT). The cauda epididymidis and vas deferens accumulated more radioactivity in progesterone and dehydroepiandrosterone (DHEA) than the caput epididymidis.
The levels of DHT, testosterone and 4-androstene-3,17-dione in the caput epididymidis were reduced after ligation of ipselateral efferent ductules indicating the testicular origin of these steroids. The cauda epididymidis and vas deferens had higher levels of progesterone as compared to the other regions of the epididymis, which were decreased after the ligation. Intravenous injection of hCG increased the levels of oestradiol-17β in all tissues and markedly in the cauda epididymidis and vas deferens. The high levels of progesterone and oestradiol-17β present in these organs may be of importance in maintaining fertilizing ability of spermatozoa stored in the cauda epididymidis and vas deferens and their transport.     

Effect of intra-epididymal injection of copper particles on fertility, spermatogenesis, and tissue copper levels in rats

In male rats, a single injection of 10 mg metallic copper particles in oil into each caput epididymidis induced infertility, whilst leaving mating behaviour and blood testosterone levels unchanged. Fertility tended to recover 5.5 months after the copper treatment. Although the copper content of the caput epididymidis reached a level of around 100 times higher than control values, the serum copper concentration did not rise significantly. This finding and the observation that treated animals gained weight as fast as the controls suggest a low systemic toxicity of this method. The testicular copper concentration was significantly higher than that in controls and different degrees of damage, including vacuolation, karyorrhexis, pyknosis, and cytolysis, were seen mainly in pachytene spermatocytes and early spermatids. Clumps of foreign particles, apparently metallic copper, were found in the interstices of the caput epididymidis together with degenerative changes in the epithelial cells of the caput, suggesting a possible effect of copper on the epididymal epithelium. The viability of epididymal sperm decreased more markedly than the decrease in sperm density. It is therefore most likely that the major cause of infertility after copper injection into the caput epididymidis is a direct inhibitory effect of copper on the sperm, whilst damage to the seminiferous and epididymal epithelial may contribute.     

Effects of cyproterone acetate on the reproductive tract and pituitary-gonadal axis of the golden hamster

Summary. The effects of cyproterone acetate (CA) on the reproductive tract and serum levels of gonadotropins and testosterone in male golden hamsters exposed to long days, were studied. The daily injection of CA at 100 mg kg⁻¹ body weight during 8 weeks reduced sperm count, both in caput and cauda epididymidis. The weight of epididymis and seminal vesicles were also reduced, while testicular weight and histology were not affected. Serum levels of testosterone and LH were increased 3.6 and 1.8 fold, respectively, above controls, while FSH was not affected by the treatment with CA. The absence of an inhibitory action of CA on the testis could be explained through an increase in LH stimulation, thus suggesting that in the golden hamster CA is devoid of progestagenic partial effects.     

Epididymal carbohydrate metabolism in experimental hypercorticosteronism: studies on mature male rats

Corticosterone induced changes in serum hormonal profiles and the key enzymes involved in glycolytic and pentose phosphate pathways were studied in caput, corpus and cauda epididymides of mature male rats (200–250 g body weight). Corticosterone (3.5 mg/100 g body weight sc. for 20 days) treatment was found to depress serum testosterone and prolactin while the gonadotrophins were unaltered. Enzymes of both the glycolytic and pentose phosphate pathways were significantly decreased in caput epididymidis. But in corpus and cauda epididymides only the glycolytic enzymes were reduced. Withdrawal of treatment (for 20 days), resulted in restoration of the glycolytic enzymes to normalcy. The serum hormonal profiles were also found to be within the normal range. The pentose phosphate pathway in caput epididymidis showed a significant increase in enzyme activities following withdrawal of treatment. From the present investigation it is clear that hypercorticosteronism had a definite influence on epididymal enzymes involved in carbohydrate metabolism.     

Effects of α-Interferon on Sperm Production, Concentration, and Motility in the Rat

Background :
We evaluated possible effects of α-interferon (α-IFN) on testicular spermatogenesis and epididymal sperm quality in the nude rat.
Methods :
Nude male rats were administered subcutaneous injections of human α-IFN daily for 3 months. The luminal content of the cauda epididymidis was collected by micropuncture. Daily sperm production was determined by Amann's method and sperm concentrations were determined by microassay. Progressive motility was judged by evaluating the linear distance traveled by the sperm in a diluent. Serum levels of testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were also measured at the end of the experiment.
Results :
Daily sperm production and epididymal sperm concentrations were significantly increased after administration of α-IFN, while progressive motility of the spermatozoa was not altered. α-IFN significantly increased serum testosterone levels, while it decreased serum LH levels and left serum FSH levels unchanged.
Conclusion :
α-IFN may improve testicular spermatogenesis and increase the epididymal sperm concentration in the rat. These promising results with α-IFN may pave the way for a new approach to treating male infertility.     

Hormonal effects on zinc concentration and morphology of rat lateral prostate gland

Hypophysectomized rats were injected with prolactin and/or testosterone, and luteinizing hormone (LH) and prolactin and/or follicle-stimulating hormone (FSH). Tissue from the lateral prostate was processed for spectrophotometric determination of zinc and for electron microscopy. Changes in zinc concentration, epithelial height, and morphological characteristics were used to assess the effect of hormonal replacement on functional activity. Although testosterone administration resulted in full restoration of all parameters, there was no evidence of a synergistic effect of prolactin and testosterone. Prolactin alone decreased the zinc concentration and increased the epithelial height compared with control animals, and there was evidence of epithelial hyperplasia. Luteinizing hormone, alone or combined with prolactin or prolactin and FSH, resulted in a marked increase of epithelial height and restoration of morphological features found in intact animals; however, only a slight increase of zinc concentration over control values was elicited. Combined LH and FSH administration reduced epithelial height and zinc concentration compared with the other LH regimes, and there was evidence of cellular degeneration.     

Zinc Content in Epididymal Spermatozoa of Metoclopramide-Treated Rats

Zinc content was determined separately in spermatozoa taken from epididymal caput and cauda in rats. It was revealed that spermatozoa transported from the epididymal caput to the cauda reduce about 54% of zinc. This reduction is significantly inhibited in spermatozoa of rats receiving metoclopramide. That is also accompanied by a fall of testosterone level in blood serum and of delta 5, 3 beta-HSD activity in Leydig cells. It was found out that the reduction of zinc in spermatozoa at the time of their passage through the epididymis is the process that depends on androgens.     

Effect of a nonsteroidal antiandrogen, flutamide on intraluminal acidification in rat testis and epididymis

Summary. Flutamide, a pure antiandrogen was administered to intact adult male rats to study the effect of altered availability of hormones on in situ pH, PCO₂ and bicarbonate concentration ([HCO⁻₃]) of seminiferous tubules, proximal caput, middle caput, middle corpus, and proximal cauda epididymidis. The weights of the epididymis and ventral prostate as well as the plasma testosterone level showed antiandrogenic effects of flutamide. Relative to controls, flutamide elevated significantly in situ pH in proximal caput, middle caput, middle corpus and proximal cauda epididymidis but not in seminiferous tubules. In situ PCO₂ values in the above segments, after flutamide, were indistinguishable from controls and from each other but all values remained significantly higher than systemic arterial blood PCO₂. Flutamide treatment did not change the [HCO⁻₃] in systemic arterial blood or seminiferous tubules but increased markedly the values in proximal caput and middle caput. The results of the present studies support the view that luminal acidification in the rat epididymis is under androgen control and may be important for sperm maturation and storage.     

Gamma-glutamyl transpeptidase activity in the epididymis during fetal and postnatal development in rats.

The histochemical and biochemical distributions of gamma-glutamyl transpeptidase (gamma-GT) were investigated in the epididymis of rats during fetal and postnatal development. In the epididymal homogenates, gamma-GT activity was detected on the fifth day after birth. A sharp increase was observed after 30 days of life in the caput homogenates. Moderate levels of the enzyme were found in the cauda epididymis. Gamma-GT is histochemically detected from the 15th day of gestation in Wolffian ducts and in 17- to 18-day-old fetuses in newly differentiated epididymal tubules. Enzyme activity, was associated with the plasma membranes (apical, lateral, and basal), was preponderant on the apical part of the epithelial cells. During the first 15 days of the postnatal life, the histochemical reaction intensities were identical from the caput to the cauda epididymidis. From the 18th day onwards, enzyme activity decreased in the corpus and in the cauda, while gamma-GT increased in the caput epididymidis, and a strong activity was found on the apical surface of epithelial cells. Weak or moderate gamma-GT activity of spermatozoa in the caput tubules, increasing steadily from caput to cauda epididymidis, suggests that gamma-GT may be related to the functional maturation of spermatozoa.     

Localization of antigenic determinants recognized by a monoclonal antibody in rat epididymal spermatozoa, with particular reference to sperm maturation

Summary.  This study localized antigenic determinants recognized by a mouse anti-human sperm monoclonal antibody TüS10 immunocytochemically and immunoelectron microscopically in the rat sperm recovered from the caput and cauda epididymidis. Immunocytochemistry showed that the antibody bound specifically to the plasma membrane overlying the principal piece of membrane-intact sperm from the caput and cauda epididymidis. Demembranation by Triton X-100 significantly decreased the affinity of the monoclonal antibody TüS10 to the caput sperm but did not obviously change that to the cauda sperm. Immunoelectron microscopy with biotinstreptavidin peroxidase complex pre-embedding method confirmed the localization of the antigenic determinants over the cell surface of the principal piece of the membrane-intact spermatozoa from the caput and cauda epididymidis. The demembranated sperm from the caput epididymidis showed no intracellular labelling, while those from the cauda displayed labelling on their external surface of the fibrous sheath. Using monoclonal antibody TüS10 as a probe, we detected different distribution patterns of the antigenic determinants between the spermatozoa in the caput and cauda epididymidis. These results suggest that spermatozoa mature with immunologically detectable changes in the fibrous sheath during their epididymal transit.     

Sperm protamine levels as indicators of fertilising potential in sexually mature male rats

We have earlier reported that administration of cyproterone acetate, fluphenazine decanoate, tamoxifen citrate, oestradiol valerate to adult male rats, at doses of 50, 5.77, 0.71, 0.28 micromol kg(-1) body weight given for periods of 15, 60, 60, 10 days, respectively, partially suppressed/reduced availability of one or more reproductive hormones viz. LH, FSH, testosterone and reduced their siring ability. The reduction in epididymal sperm counts was not considerable after treatment with these drugs, but conventional methods of assessment of spermatozoa quality viz. sperm chromatin structure assay (SCSA), nuclear chromatin decondensation (NCD) assay, monobromobimane (mBBr) uptake, had shown quantifiable changes in caput sperm chromatin compaction and reduced the testicular levels of protamine 1. The present follow-up study attempts to quantify changes in caudal sperm chromatin which has undergone compaction in the epididymis, in the altered hormonal microenvironment of rats treated with cyproterone acetate, tamoxifen citrate, fluphenazine decanoate, oestradiol valerate, at doses of 50, 5.77, 0.71, 0.28 micromol kg(-1) body weight respectively given for periods of 15, 60, 60, 10 days, with a view to correlating these changes to reduction in their fertilising potential. During the androgen-dependent transit of spermatozoa from caput to cauda epididymis, thiol group oxidation and tyrosine phosphorylation of protamine occurs in maturing sperms concomitant with development of fertilising ability. The results indicate that conventional methods viz. SCSA, NCD, mBBr uptake fail to detect changes induced by hormone deficits in sperm chromatin condensation, as a result of maturation during transit from caput to cauda epididymis. Absence of protamine 1 in epididymal sperm was observed in either testosterone or FSH deficient rats that correlated with reduced fertilising potential. The study suggests that changes in LH/T or FSH affect a hitherto unknown common molecular mechanism in the testis, underlying the protamination of rat spermatozoa. In conclusion, loss of P1 occurs in adult male rats deprived of T or FSH and is a reliable detectable change in epididymal sperm indicative of chromatin condensation defects associated with endocrine imbalance and poor fertility status.     

Effect of testicular capsulotomy on secretion of testosterone and gonadotrophins in rats

Aim: In order to clarify further the mechanisms underlying the effect of capsulotomy on testicular function, the levels of testosterone, LH and FSH were observed. Methods: Intratesticular testosterone levels and LH, FSH levels in the peripheral blood of normal, sham-operated and capsulotomized rats were detected by RIA. Results: After testicular capsulotomy, there was a progressive reduction in the testosterone level in the testicular venous blood together with a progressive increase in the LH and FSH levels in the peripheral blood from approximately 30 days post-capsulotomy. Morphological changes were observed at 5-10 days after capsulotomy, i.e., far ahead of the hormonal changes.Conclusion: The seminiferous tubular damage after testicular capsulotomy was not caused by the reduction in testosterone, and on the contrary, the hormonal change might be secondary to the morphological alterations. The increase in LH level most likely resulted from a negative feedback influence from the lowered testosterone level, while the increase in FSH secretion may be a feedback signal of the damaged seminiferous tubules. (Asian J Androl 2000 Dec;2:257-261)     

Glutathione peroxidase activity in cell cultures from different regions of human epididymis

AIM: To study the secretory activity and androgen regulation of glutathione peroxidase (GPx) in epithelial cell cultures from human epididymis. METHODS: Tissue was obtained from patients undergoing therapeutic orchidectomy for prostatic cancer. Epithelial cell cultures were obtained from the caput, corpus and cauda epididymides. Enzymatic activity was measured in conditioned media by colorimetric methods in absence or presence of 1, 10 or 100 nmol/L testosterone. The effect of 1 micromol/L flutamide was also evaluated. RESULTS: GPx activity was higher in cultures from corpus and cauda than caput epididymidis. The presence of different concentrations of testosterone increase enzyme activity in cell cultures from all epididymal regions. Addition of flutamide reverses the androgen dependent increase of GPx activity. CONCLUSION: GPx activity is secreted from human epididymal cells in a region dependent manner and is regulated by androgens.     

Loss of sperm surface sialic acid induces phagocytosis: an assay with a monoclonal antibody T21, which recognizes a 54K sialoglycoprotein

Using a monoclonal antibody T21, we reported that a mouse sperm maturation-associated antigen sialoglycoprotein of 54000 daltons (54K sialoglycoprotein) was secreted at the distal caput to proximal corpus epididymidis and that the 54K sialoglycoprotein had a hidden determinant (cryptodeterminant), which could be eliminated by sialidase treatment (Toshimori et al. (1988): Histochemistry 90:195-200; (1990a): Biol Reprod 42:151-160; (1990b): Arch Histol Cytol 53:339-349). This study evaluated the mouse sperm susceptibility to phagocytosis by macrophage in vitro. Comparisons were made between sperm from the caput epididymidis (caput sperm) incubated in modified Krebs Ringer's solution (MKR) and caput sperm incubated in MKR containing cauda fluid, and between sialylated (sialidase-untreated) sperm from the corpus and cauda epididymidis (corpus/cauda sperm) and desialylated (sialidase-treated) corpus/cauda sperm. The results showed that macrophages were least actively engaged in phagocytosis for caput sperm incubated in MKR containing cauda fluid, and most active for desialylated corpus/cauda sperm. Incubation of caput sperm in MKR containing cauda fluid revealed that the 54K sialoglycoprotein in cauda fluid could be bound to the flagellar surface of caput sperm. These results together with previous findings strongly suggest that the 54K sialoglycoprotein bound to immature sperm during maturation in the epididymis is implicated in the protection of sperm from phagocytosis with the aid of sialic acid residues.     

Endocrine Effects of Early Non-Union of Testis and Epididymis and of Cryptorchidism in the Rat/Endokrine Effekte bei jungen Ratten nach experimenteller Trennung von Hoden und Nebenhoden wie auch nach experimentellem Kryptorchismus

The aim of this study was to explore possible endocrine effects of early non-union of testis and epididymis. In 16 days old Sprague-Dawley rats the testis and epididymis were separated to the level of the inferior epididymis artery (non-union operation). Animals were killed at intervals from 30-58 days, and the plasma concentrations of total testosterone, total estrogens, LH, FSH and PRL were measured. The testes were studied by light microscopy. Groups of rats made cryptorchid and sham-operated rats were used as controls. Although their testes were scrotal, the non-union operated animals had testosterone and estrogen concentrations similar of cryptorchid animals, and significantly (p less than 0.05) lower than sham-operated animals. The LH- and FSH-concentrations were significantly (p less than 0.05) elevated suggesting a primary lesion in both Leydig- and Sertoli-cells. Towards puberty FSH increased in the non-union operated animals, while FSH values declined in the cryptorchid and sham-operated animals. These FSH-patterns probably reflect the existence of different pathogenic mechanisms in the non-union operated rats and the rats with cryptorchid testes.