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PubMed1987

Tetracycline sclerotherapy for hydroceles and epididymal cysts.

Tetracycline (Achromycin, Lederle) solution was used to produce sclerosis of 37 hydroceles and 18 epididymal cysts in patients over the age of 50 years. All but one of the epididymal cysts were sclerosed satisfactorily at 9 months; 35 hydroceles sclerosed satisfactorily, although four patients required re-treatment at 3 months. One patient required orchiectomy for an infected haematocele. Pain after instillation of tetracycline was an occasional complication which could be prevented by prophylactic spermatic cord block. Tetracycline sclerotherapy is a safe and effective alternative to surgery for hydroceles and epididymal cysts in older patients.

Aged↗
PubMed1988

Ethanolamine oleate as a sclerosant for testicular hydroceles and epididymal cysts.

Forty patients with testicular hydroceles and 16 patients with epididymal cysts were treated by tapping and injection of ethanolamine oleate; 97.5% of the hydroceles and 31% of the epididymal cysts were completely cured, 50% of the latter partly cured and 19% failed. About half of the patients experienced pain after treatment. Two patients developed infection. Ultrasonography showed no change in the testicular tissue during the follow-up period. Sclerotherapy with ethanolamine oleate is a rapid and cost-effective modality which can be recommended as primary treatment for hydroceles and for unilocular epididymal cysts, especially in elderly males.

Adult↗
PubMed1995

Anti-bacterial activity of the fluid contents of spermatoceles and epididymal cysts.

OBJECTIVE: To show that the fluids obtained from spermatoceles and epididymal cysts are not infected, even though they may be present for long periods, and that these fluids have bactericidal activity. MATERIALS AND METHODS: Sixteen patients, 13 with a spermatocele and three with an epididymal cyst, were included in the study. Protein, glucose, chloride, inorganic phosphorus, calcium and magnesium contents were measured and cultures of the fluids were carried out. Bactericidal activity against the Escherichia coli NTCC 10418 clone was tested in different dilutions. RESULTS: Biochemical analysis showed that the protein, glucose and ionic content of the fluids was lower than that of serum, except for chloride. Microbiological cultures were negative for all samples. A significant bactericidal effect was obtained with 1/1 dilution and no reproduction was seen with this dilution. CONCLUSION: These findings indicate that the fluids within spermatoceles and epididymal cysts do not become infected under normal circumstances.

Adult↗
PubMed2004

Should cryopreserved epididymal or testicular sperm be recovered from obstructive azoospermic men for ICSI?

OBJECTIVE: To determine the effect of the anatomical site of sperm recovery on intracytoplasmic sperm injection (ICSI) embryo implantation, pregnancy and live birth rates in couples with isolated obstructive azoospermia as the sole cause of infertility. DESIGN: Controlled, single centre, retrospective clinical study. SETTING: University Hospital, Centre for Reproductive Medicine. SAMPLE: One hundred and fifty-one cycles of ICSI were performed, using surgically recovered sperm, between August 1996 and March 2002. METHODS: The outcome of ICSI, with surgically recovered sperm, was compared between epididymal (Group E) and testicular (Group T) derived sperm. Inclusion was limited to couples undergoing their first treatment cycle, where female age was < or =39 years and a minimum of five oocytes were available for injection. Women with a history of ovarian surgery, ultrasonic evidence of polycystic ovaries, uterine anomalies or hydrosalpinx were excluded. MAIN OUTCOME MEASURES: Clinical pregnancy, implantation and live birth rate. RESULTS: Forty-two of 151 cycles met the strict inclusion criteria. Groups E and T were comparable with respect to age, basal serum FSH, ovarian response; number of oocytes injected and number of embryos available and transferred. No difference existed between Groups E and T in implantation, clinical pregnancy or live birth rate (28.8% vs 25.8%, 42.9% vs 42.9% and 39.3% vs 42.9%, respectively). CONCLUSIONS: Cryopreserved epididymal and testicular sperm, from men with obstructive azoospermia, appear equally effective in ICSI. Epididymal recovery should remain the method of first choice for obstructive azoospermic men but further study of sperm DNA damage rates in different testicular sites is required.

Adult↗
PubMed1979

The effects of alpha-adrenoceptor agonists and antagonists on responses of transmurally stimulated prostatic and epididymal portions of the isolated vas deferens of the rat.

1 The effects of alpha-adrenoceptor agonists and antagonists on contractile responses of transmurally stimulated prostatic and epididymal portions of the rat isolated vas deferens were examined. 2 Responses to single stimuli consisted of two phases, the first predominant in the prostatic and the second in the epididymal portion. The first phase was resistant to alpha-adrenoceptor antagonists but the second was reduced in a dose-related manner in the order of potency prazosin greater than azapetine greater than phentolamine greater than labetalol greater than yohimbine. 3 Both phases of the response to a single stimulus were reduced by clonidine but only the first could be reliably restored by yohimbine. 4 Trains of transmural stimuli produced biphasic responses, an early rapid component predominant in the prostatic and a slow secondary component predominant in the epididymal portion. The effects of alpha-adrenoceptor antagonists on these responses were complex. Prazosin produced the most straightforward inhibition of responses with relative resistance of the early rapid component. Only yohimbine and phentolamine produced increases in responses which could be pre-junctional in origin. 5 The alpha-adrenoceptor agonists, oxymetazoline and clonidine, reduced while phenylephrine increased responses to trains of stimuli. 6 These results are discussed in relation to the nature of the innervation of rat vas deferens and the usefulness of the preparation in pharmacological tests for activity at alpha-adrenoceptors.

Adrenergic alpha-Agonists↗
PubMed1995

Evidence for a functional alpha 1A- (alpha 1C-) adrenoceptor mediating contraction of the rat epididymal vas deferens and an alpha 1B-adrenoceptor mediating contraction of the rat spleen.

1. The alpha 1-adrenoceptor subtype mediating contraction of the rat epididymal vas deferens and rat spleen has been investigated by use of alpha 1-adrenoceptor antagonists that have shown selectivity between the different cloned receptor subtypes. 2. In the rat epididymal vas deferens the potency of noradrenaline and phenylephrine was increased in the presence of neuronal and extra-neuronal uptake blockers, cocaine and beta-oestradiol, but these did not alter that of methoxamine. The order of potency of the agonists in the presence or absence of uptake blockade was noradrenaline > phenylephrine > methoxamine. In the rat spleen the potency of these agonists was not altered in the presence of cocaine and beta-oestradiol, and their order of potency was the same as in the vas deferens. 3. The non subtype selective alpha 1-adrenoceptor antagonist prazosin (up to 1 x 10(-7) M) was found to antagonize contractions to noradrenaline in the vas deferens competitively (pA2 9.2), but only in a non competitive manner in the spleen. Contractions to phenylephrine in the spleen however were competitively antagonized by prazosin (up to 1 x 10(-7) M) with a pA2 of 9.2. This suggests that there is an alpha 1- and a non alpha 1-adrenoceptor response to noradrenaline in the rat spleen. 4. Pretreatment with chlorethylclonidine (10(-4) M for 30 min) did not alter the noradrenaline contractions in the vas deferens, but contractions to noradrenaline and phenylephrine in the spleen were shifted 30 and 300 fold to the right of the control curve, respectively. This suggests that only the contractions in the spleen were mediated by alpha 1B-adrenoceptors. 5. The noradrenaline contractions in the vas deferens were competitively antagonized by WB 4101 (pA29.6), 5-methyl-urapidil (pA2 8.7), phentolamine (pA2 8.3), benoxathian (pA2 9.4), spiperone (pA2 7.5),indoramin (pA2 8.4) and BMY 7378 (pA2 6.7), consistent with the affinities of these antagonists in binding studies on tissue alpha 1A-adrenoceptors. These values correlated best with their published affinities on the expressed alpha 1c-adrenoceptor clone and poorly with those at either the expressed alpha lb- or alpha 1d adrenoceptor clones. Therefore the classical alpha 1A-adrenoceptor appears to be the same as the expressed alpha lc-adrenoceptor clone.6. The phenylephrine contractions in the spleen were competitively antagonized by WB 4101 (pA2 8.1),5-methyl-urapidil (pA2 7.1), phentolamine (pA2 7.3), benoxathian (pA2 7.4), spiperone (pA2 7.9),indoramin (pA2 7.5) and BMY 7378 (pA2 7.4), consistent with the affinities of these antagonists in binding studies on tissue alB-adrenoceptors. The pA2 values correlated best with the published affinities of these compounds on the expressed alb-adrenoceptor clone and poorly with those at either the expressed alpha ld- or alpha lc-adrenoceptor clones. Therefore the alpha lB-adrenoceptor appears to be the same as the expressed alpha lb-adrenoceptor clone.7. The results provide pharmacological evidence that the alpha1-adrenoceptor mediating noradrenaline contractions in the epididymal portion of the rat vas deferens is the alpha 1A-(alpha lC) subtype and that contractions to phenylephrine in the rat spleen are mediated by the alpha 1B-subtype.

Adrenergic alpha-Agonists↗
PubMed2006

Can inhibin-B predict the outcome of microsurgical epididymal sperm aspiration in patients with suspected primary obstructive azoospermia.

AIM: To evaluate whether inhibin-B can predict the outcome of a microsurgical epidymal sperm aspiration (MESA) procedure in patients with suspected primary obstructive azoospermia (OA) and if inhibin-B can replace testicular biopsy in the diagnostic work-up of these patients. METHODS: Inhibin-B levels and testicular biopsy scores were related to the outcome of MESA in 43 patients with suspected primary OA. MESA was considered to be successful when epididymal sperm could be identified during the procedure. RESULTS: Spermatozoa were present in the epididymal aspirate in 28 out of the 43 patients (65%). Inhibin-B values were not significantly different in patients with successful or unsuccessful MESA. The modified Johnsen score, however, was significantly lower in patients with unsuccessful MESA (P = 0.003). A rete testis obstruction or epididymal malfunctioning was found in 15% of patients with suspected primary OA, reflected by unsuccessful MESA despite normal inhibin-B levels and normal testicular histology. CONCLUSION: Inhibin-B cannot replace testicular biopsy as a diagnostic tool in the work-up of patients with suspected primary OA. Testicular biopsy is useful in identifying patients with spermatogenic arrest, who might have normal inhibin-B values.

Azoospermia↗
PubMed1990

Secretory agonists stimulate a rise in intracellular cyclic AMP but not Ca2+ and inositol phosphates in cultured rat epididymal epithelium.

Primary monolayer cultures of rat epididymal cells have been shown to secrete chloride and bicarbonate when stimulated with beta-adrenergic agents, humoral agents and vasoactive peptides. The intracellular messengers mediating the secretory response are unknown. In this study intracellular AMP, Ca2+ and inositol phosphates were measured in epididymal monolayers at rest and upon stimulation with various secretory agonists. Adrenaline, forskolin, lysylbradykinin, prostaglandin, endothelin, angiotensin II, antidiuretic hormone and vasoactive intestinal peptide at concentrations that stimulate anion secretion caused a rise in intracellular cyclic AMP. The increase in cyclic AMP by adrenaline was blocked by propranolol but not by phentolamine. Studies of the concentration-effect relationships showed that for adrenaline and endothelin the EC50 for stimulation of cyclic AMP was higher than that for stimulation of anion secretion. None of these agonists affects intracellular Ca2+ concentration and inositol phosphate contents in epididymal monolayers. Ca2+ ionophores A21387, ionomycin and erythrosin B (with irradiation with white light), at concentrations that stimulate anion secretion, also stimulated a rise in intracellular cyclic AMP and concomitantly increased intracellular Ca2+. The increase in cyclic AMP was dependent on extracellular Ca2+. It is not known whether the secretory response to Ca2+ ionophores was mediated by an increase in cell Ca2+ per se, or cyclic AMP. However, it can be concluded that cyclic AMP is the second messenger which mediates the secretory responses to physiological stimuli.

Adrenergic Agonists↗
PubMed1994

Swelling-induced anion and cation conductances in human epididymal cells.

1. Activation of both anion and cation conductances was observed in primary cultured human epididymal cells during osmotic swelling under the patch-clamp whole-cell configuration. The swelling-induced anion conductance was 25.66 +/- 4.70 nS and the cation conductance was 7.35 +/- 1.40 nS. The permeability ratio of K+ to Cl- (PK/PCl) was calculated to be 0.40. Known anion or cation channel blockers could inhibit both conductances simultaneously. 2. When the major permeant ion species in the pipette and bath solution was Cl-, the mean conductance was found to be 17.06 +/- 1.8 nS, significantly smaller than that obtained in the presence of intracellular K+, 25.66 +/- 4.70 nS (P < 0.05). No significant current activation was observed when solutions containing only K+ as the permeant ion were used. 3. When the anionic amino acids glutamate and aspartate were used to replace extracellular Cl-, the permeability ratios were calculated to be PGlut/PCl = 0.20 and PAsp/PCl = 0.17. 4. The cation conductance was found to be non-selective since its permeability to other cations such as Na+ and choline, an organic compound highly concentrated in epididymal fluid, was similar to that of K+. 5. Regulatory volume decrease (RVD) was observed after initial osmotic swelling; this could be inhibited by either anion or cation channel blockers. 6. The results of this study suggest that both anion and cation conductances are activated during cellular swelling, and indicate the existence of an interdependent relationship between the swelling-induced cation and anion conductances. Both swelling-induced cation and anion conductances are involved in the volume regulatory process and may be responsible for transporting amino acids or organic compounds in human epididymal cells.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1996

Cyclic AMP phosphodiesterase: a regulator of forward motility initiation during epididymal sperm maturation.

The concentrations of cAMP, cAMP phosphodiesterase (PDE) activity, and the effect of theophylline in vitro on the forward motility (FM) of maturing goat epididymal sperm have been analyzed. cAMP levels increase slowly during transit of the cells from the caput to the proximal cauda, although they acquired a minimal degree of forward progression. The last phase of sperm transit (proximal to distal cauda) was associated with a concomitant sharp rise in the level of both cAMP as well as flagellar motility. PDE activity progressively decreased (approximately threefold) during epididymal maturation, being minimal in mature cauda sperm. Theophylline (30 mM), a specific inhibitor of PDE, markedly activated (10-fold or greater) motility of the sperm derived from proximal-corpus, mid-corpus, distal-corpus, and proximal-cauda epididymides. FM of the native mature caudal sperm was similar to that of the theophylline-treated proximal-cauda sperm. The terminal stage of sperm maturity (proximal to distal cauda) was associated with a markedly reduced level of theophylline-dependent motility activation (approximately 50%). The data are consistent with the view that PDE plays an important role in the initiation of motility during epididymal sperm maturation.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1994

Effects of hochu-ekki-to, a Japanese kampo medicine, on cultured hamster epididymal cells.

In Japan, Hochu-ekki-to (TJ-41), a Japanese Kampo preparation is used extensively in the treatment of idiopathic male infertility. In order to elucidate the mechanism of how this drug affects spermatogenesis, we examined the effects of the sera from male mice to which TJ-41 was administered orally, on protein synthesis in cultured hamster epididymal cells. Golden hamster epididymal cells were cultured in RPMI1640 medium supplemented with FCS for 7 to 10 days. After the initial culture period, the medium was changed to RPMI1640 supplemented with 1 microCi/ml [3H]-leucine and 10% serum from male ICR mice to which TJ-41 was orally administered for 7 days. The culture was then continued for 20 hours, and the uptake of [3H]-leucine into the cultured hamster epididymal cells was measured. The uptake of [3H]-leucine was significantly higher (p < 0.05) in cells cultured in media supplemented with sera from the TJ-41 treated mice than in cells cultured with control sera. Elements in the sera from control and TJ-41 treated mice were analyzed by high performance liquid chromatography (Waters HPLC, type 510; Milipore) with a mu-Bondapack C18 column. Several new peaks were detected in the sera from TJ-41 treated mice. These results and the clinical data suggest that TJ-41 may promote the synthesis of several proteins which might he related to the functional maturation of spermatozoa in the epididymis.

Administration, Oral↗
PubMed1993

ATP stimulates Ca2+ release from a rapidly exchanging pool in cultured rat epididymal cells.

A study was carried out to investigate an ATP-sensitive Ca2+ pool in rat epididymal cells and its role in transepithelial Cl- secretion. In normal buffered solution containing 2.5 mM free Ca2+, ATP triggered single calcium spikes in a dose-dependent fashion. In nominally Ca(2+)-free solution, the peaks of successive Ca2+ spikes diminished after repeated ATP stimulations. Addition of Sr2+ (2.5 mM) to Ca(2+)-free solution after ATP stimulation did not cause changes in fluorescence signals. However, in the presence of Sr2+, ATP gave rise to apparent repetitive Ca2+ spikes of similar magnitudes after repeated stimulations. Increasing the time of exposure in Ca(2+)-free solution containing 50 microM ethylene glycol-bis(beta-amino-ethyl ether)-N,N,N',N'-tetraacetic acid rapidly decreased the intracellular Ca2+ concentration ([Ca2+]i) response to subsequent ATP stimulation. On the other hand, increasing the time of exposure in Sr(2+)-containing solution in Ca(2+)-depleted cells rapidly increased the apparent [Ca2+]i response to subsequent ATP stimulation. These observations suggested the existence of a Ca2+ pool that was rapidly exchanging with the extracellular compartment. Apical application of ATP elicited a transient rise in short-circuit current across the epididymal epithelium in a dose-dependent fashion, and the response was reduced by prior stimulation with thapsigargin. Ca2+ released from a rapidly exchanging ATP-sensitive store might stimulate Cl- secretion in the epididymis, thereby maintaining the electrolyte contents and fluidity of the epididymal microenvironment.

Adenosine Triphosphate↗
PubMed1975

Influence of age on size and number of fat cells in the epididymal depot.

The extensive literature on the effect of rat age on the size and number of adipocytes in adipose tissue depots relates solely to young developing rats and young adults. Therefore a study was carried out in our laboratory on the cellular characteristics of the epididymal depot of the Fisher 344 strain of rats through virtually the entire life-span. Collagenase digests of this depot prepared from rats of 9, 13, 26, 52, 104, and 130 wk of age yield a population of cells with diameters greater than 30 mum identified as adipocytes or "fat cells." A remarkably complex pattern of changes in both the size and the number of these fat cells in the epididymal depot occurred through the life-span of the rats. The epididymal depots also contain some cells with diameters around 10 mum which have a morphology similar to that of the classic adipocytes; such cells may be preadipocytes.

Adipose Tissue↗
PubMed1988

Dietary influence on the insulin function in the epididymal fat cell of the Wistar rat. III. Effect of the ratio carbohydrate to fat.

Newly weaned male Wistar rats were fed semipurified diets containing sunflowerseed oil or palm oil. The fatty acid compositions of the phospholipid and triacylglycerol fractions either from whole epididymal fat pads or from fat cells isolated from the pads appeared to be nearly similar. Feeding the diets for a period of only 3-4 weeks after weaning was found to be sufficient to produce a difference in insulin response and insulin binding in the epididymal fat cells. A reduction in the dietary fat content from 30 to 15% of the energy had only minor effects on fat pad weight and on the fatty acid composition of the phospholipid- and triacylglycerol fractions isolated from the epididymal fat pads. There was no effect either on body weights or on average fat cell diameter but there was an increase in insulin response in the isolated fat cells, however. The difference in insulin response after a diet rich in polyunsaturated fatty acids (sunflowerseed oil) as compared to one rich in saturated fatty acids (palm oil) remained after the reduction of fat content.

Adipose Tissue↗
PubMed1989

Epididymal interstitial (Leydig) cell tumors in B6C3F1 mice.

Six primary interstitial cell tumors of the epididymis were identified from 46,752 male B6C3F1 mice used in chronic toxicity and carcinogenicity studies. Five of the tumors occurred at the end of 2-year studies; none were attributed to treatment. None of the mice with epididymal tumors had a primary testicular tumor. Histologically, tumors were characterized by a nodular or diffuse proliferation of tumor cells in the epididymal interstitium. Most cells were polygonal with highly vacuolated cytoplasm (vacuolated cells) or eosinophilic cytoplasm (eosinophilic cells). Smaller hyperchromatic cells with scant basophilic cytoplasm (basophilic cells) and cells with yellow-brown pigment characteristic of lipofuscin (pigmented cells) were less common. In each tumor two or more cell types were present. Extension of these tumors through the capsule, invasion of the testis, or metastasis did not occur. By electron microscopy both eosinophilic and vacuolated cell types had a large round or oval nucleus with sparse heterochromatin, abundant mitochondria with tubulovesicular cristae, and frequent desmosome structures between cell membranes. Vacuolated cells contained numerous lipid droplets. Morphological features of the epididymal tumors are similar to those of the testicular interstitial (Leydig) cell tumor in mice and rats.

Animals↗
PubMed2003

Identification, immunolocalization, regulation, and postnatal development of the lipocalin EP17 (epididymal protein of 17 kilodaltons) in the mouse and rat epididymis.

Several lipocalins are present in the mouse epididymis and are thought to play a role in sperm maturation by transporting lipophilic molecules. We have previously reported that two lipocalin genes, mERABP (mouse epididymal retinoic acid binding protein), and mEP17 (mouse epididymal protein of 17 kDa), derived from an ancestral gene, are specifically expressed in the epididymis. In the present study, a polyclonal antibody was raised against a recombinant protein to investigate the presence and the regulation of mEP17. mEP17 was detected in the supranuclear region of the principal cells of the initial segment, the clear cells of the caput epididymidis, and the lumen of the mid/distal caput but not of the distal epididymis. Initial segment and caput tissue extracts were subjected to HPLC separation. After electrophoresis of the immunoreactive mEP17-enriched fractions, the immunoreactive band was analyzed by mass spectrometry to identified mEP17 unambiguously. After two-dimensional electrophoresis, mEP17 appeared as a train of five 22-kDa spots with a range of pI (isoelectric point) from 5.8-6.7. N-glycanase digestion gave rise to a single spot of 17 kDa and pI 6, the predicted mass and pI. During ontogeny, mEP17 was detected as early as 3 wk of age and increased afterward. After bilateral orchiectomy, mEP17 disappeared 2 d after surgery and was not restored after testosterone replacement. After unilateral orchiectomy, mEP17 levels decreased only in the orchiectomized side. After cryptorchidism or busulfan treatment, mEP17 levels were either greatly diminished or not detected. This suggests that mEP17 is dependent on testicular factor(s) that may have a germ cell origin. Altogether, our data demonstrate that mEP17 spatial expression, regulation, and fate are different from that of the highly related mouse epididymal retinoic acid binding protein. This suggests that these two related proteins exhibit distinct functions in the mouse epididymis.

Aging↗
PubMed1987

Analysis of the disruptive action of an epididymal protease and the stabilizing influence of molybdate on nondenatured and denatured glucocorticoid receptor.

The nucleomyofibrillar fraction of epididymides from sexually mature rabbits contains a novel leupeptin-sensitive protease that disrupts the oligomeric conformation of cytosolic estrogen and progesterone receptors, and molybdate inhibits this process. In this report we used the AtT-20 cell glucocorticoid receptor as substrate and performed analyses under nondenaturing vs. denaturing conditions to further investigate the effects of the epididymal protease and molybdate on steroid receptor structure. Analysis on low salt sucrose gradients indicated that the protease partially converted the oligomeric (9-10S) glucocorticoid receptor to several more slowly sedimenting forms (3-7S), and this effect was not observed in the presence of molybdate. Paradoxically, gradient analysis under high salt conditions revealed that the protease induced a discrete, quantitative and molybdate-insensitive conversion of the 4-5S steroid-binding subunit to a 3S form. Further studies were done using denaturing polyacrylamide gel electrophoretic analysis of receptor that had been labeled covalently with [3H]dexamethasone 21-mesylate and partially purified by DNA/cellulose chromatography. At 0-4 C, the protease cleaved the steroid-binding subunit (mol wt, 96,900) of the receptor to a single steroid-labeled fragment (mol wt, 42,600). Under these conditions, digestion was complete within 30-60 min and was inhibited by leupeptin, but was unaffected by thiol-reactive reagents or molybdate. The epididymal protease and alpha-chymotrypsin produced steroid-labeled receptor fragments that were indistinguishable in size, shared an epitope recognized by our BuGR-2 monoclonal antibody, and retained DNA-binding activity. Despite the apparent similarity of these two enzymes, they are distinct, since the chymotrypsin-dependent cleavage event was not inhibited by leupeptin. These studies show that the epididymal protease attacks a site on the steroid-binding subunit of glucocorticoid receptors as well as estrogen and progestin receptors. It also appears that the cleavage site is situated close to that most readily attacked by alpha-chymotrypsin. Finally, our data provide independent confirmation of a recent report indicating that molybdate ions interact directly with the cytosolic steroid receptor to stabilize its oligomeric structure even after proteolysis within the steroid-binding subunit.

Animals↗
PubMed1992

Evidence that rodent epididymal sperm contain the Mr approximately 94,000 glucocorticoid receptor but lack the Mr approximately 90,000 heat shock protein.

Monoclonal antibodies directed against four different polypeptide epitopes on the Mr approximately 94,000 steroid-binding subunit of the rat liver cytosolic glucocorticoid receptor (GcR) were used to probe Western blots of epididymal spermatozoa from rats and mice. Two sperm polypeptides with apparent molecular weights of 94,000 (indistinguishable in size from the liver GcR subunit) and 150,000 reacted with these antibodies. Other polypeptides that are present in a wide variety of somatic cells [lamin-A, -B, and -C; topoisomerase-I; poly(ADP-ribose) polymerase; the 62-kilodalton internal nuclear matrix protein; the nucleolar protein B23; and histone H1] could not be detected in these preparations of spermatozoa, thus appearing to rule out contamination by somatic cells. Rat and mouse pachytene spermatocytes and round spermatids contained much lower amounts of the Mr approximately 94,000 and 150,000 polypeptides. These results suggested that the steroid-binding subunit of the GcR might be accumulated late in spermatogenesis. Consistent with this view, a 6-kilobase mRNA (identical in size to a mRNA detected in mouse somatic cell lines) was detected when Northern blots of mouse round spermatid RNA were probed with a cDNA to the steroid-binding GcR subunit. Although the results described above suggest the presence of GcR in rodent sperm, high affinity binding of glucocorticoids to epididymal sperm could not be detected in a whole cell binding assay. Further analysis revealed that the Mr approximately 90,000 heat shock protein (hsp90), a component reportedly required for high affinity ligand binding to the GcR, was present in early germ cells, but absent from rodent epididymal sperm. These results suggest that the Mr approximately 94,000 steroid-binding subunit of the GcR and an immunologically related Mr approximately 150,000 polypeptide are specifically accumulated during the later stages of rodent spermatogenesis, but are not assembled into receptor complexes capable of binding steroid. In addition, these results support the view that hsp90 is required for high affinity binding of glucocorticoids to the Mr approximately 94,000 GcR subunit in intact cells.

Adrenalectomy↗