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Showing papers in "Journal of Andrology in 1983"


Journal ArticleDOI
TL;DR: The results suggest that, compared to the mouse, spermatogenesis in man is approximately 3.1 times more sensitive to ionizing irradiation.
Abstract: Existing data concerning the effects of x-irradiation on spermatogenesis in man were analyzed and the results were compared to published data on the mouse. Testicular x-irradiation produced a transient, but substantial, suppression of sperm counts in man, with an ED50 near 11 rad. The length of time to recovery was proportional to the irradiation dose. The ED50 for suppression of type A spermatogonia following radiation exposure in man was similar (9.7 rad), although the response curves for spermatogonia and sperm count were matogonia in the mouse was parallel to that found in man, but with an ED 50 of 30.0 rad. These results suggest that, compared to the mouse, spermatogenesis in man is approximately 3.1 times more sensitive to ionizing irradiation.

200 citations


Journal ArticleDOI
TL;DR: As a single parameter, decreased number of normal forms appears to be a good indicator for clinical infertility if in vitro fertilization testing is not available.
Abstract: Semen samples from 95 men were examined by routine semen analysis and specific histologic staining for sperm morphology. The men were classified into fertile and infertile groups on the basis of clinical evaluation and in vitro testing, using the zona-free hamster egg penetration assay. Thirty men were designated as fertile, as they had fathered children and their sperm showed penetration of greater than 20% of the zona-free hamster eggs with the in vitro fertilization test. Subjects classified as infertile were men from infertile couples whose wives showed no evidence of infertility and whose in vitro fertilization ability was 10% or less. The semen analysis parameters of the fertile and infertile groups were significantly different. Fertile men had mean values of 108 X 10(6) sperm/ml, 61% motile, 64% normal forms (sperm with oval morphology), and 69% penetration in vitro. The mean values for infertile men were significantly lower: 42 X 10(6) sperm/ml, 45% motile, 32% normal forms, and 3.2% penetration in vitro. The importance of the morphology parameter was revealed by comparison of the percentage of penetration with count, motility, and morphology. Penetration correlated best with morphology (r = 0.730) as compared with motility (r = 0.451) and count (r = 0.605). The distribution of abnormalities in the infertile group revealed 81.6% with abnormal morphology (less than 50%), 53.8% with abnormal motility (less than 50%), and 38.5% with abnormal count (less than 20 million/ml). As a single parameter, decreased number of normal forms appears to be a good indicator for clinical infertility if in vitro fertilization testing is not available.

105 citations


Journal ArticleDOI
TL;DR: The collected data indicate the absolute need for adequate standardization of this important test, which is the basis upon which the clinician makes the important decision of implicating the male partner in the couple's infertility problem.
Abstract: Sixty-four laboratories from Connecticut, Massachusetts, and California were involved in a comparison of the types of semen analyses offered. The collected data indicate that there is a wide range of normal values for each parameter considered in a semen analysis. Very few laboratories gave simple and precise instructions for collection of the specimen. In many instances there was no record of the collection time or of the arrival time of the specimen at the laboratory. Most laboratories did not report motility or forward progression at time intervals, and abnormal morphology was not broken down according to the types of abnormalities involved. These data indicate the absolute need for adequate standardization of this important test, which is the basis upon which the clinician makes the important decision of implicating the male partner in the couple's infertility problem.

63 citations


Journal ArticleDOI
TL;DR: The results demonstrate rapid changes in aspects of both Sertoli and Leydig cell function after surgically-induced cryptorchidism and a highly negative correlation between the rise and both ABP and fluid production.
Abstract: The effects of surgically-induced cryptorchidism on testicular function in adult rats was studied, with particular emphasis on the temporal relationship between the onset of cryptorchidism and a number of parameters of Sertoli cell, Leydig cell, and pituitary function. Sertoli cell function assessed by measurement of testicular ABP content, ABP production rate, testicular fluid production, and FSH receptor levels was found to be disrupted very rapidly (one to two days) after inducing cryptorchidism. By four days, ABP production rate and fluid production were found to represent only 12% and 65% of control levels, respectively. However, these parameters were maintained at these levels even 70 days following cryptorchidism. The binding of 125I-FSH to testes homogenates was found to decrease steadily to less than 10% of control levels after 70 days of cryptorchidism. Serum FSH levels were elevated 14 days postcryptorchidism and there was a highly negative correlation between the rise and both ABP and fluid production. Serum levels of LH rose after 14 days of cryptorchidism, although testosterone levels were not significantly altered throughout the experiment. The binding of iodinated hCG to testes homogenates also decreased significantly within six days of inducing cryptorchidism and remained low (16%) after 70 days of cryptorchidism. The results demonstrate rapid changes in aspects of both Sertoli and Leydig cell function after surgically-induced cryptorchidism.

59 citations


Journal ArticleDOI
TL;DR: The present data suggest that, in unselected male patients seeking reproductive evaluation, the sperm penetration assay did not yield any additional information on the cause of infertility.
Abstract: Sperm penetration rates in the heterologous ovum penetration test were correlated with results of routine semen analysis in 30 fertile and 50 infertile men. There was no difference in penetration rates when comparing infertile men with normal and abnormal seminal parameters, nor was any difference seen between fertile (15-83%) and infertile men (8-83%). Of the 22 infertile men with normal seminal characteristics, seven had partners with no discernible reproductive dysfunction. The penetration rates of these men (38-81%) did not differ from those of fertile men. Stepwise regression analysis of seminal characteristics, with penetration as the dependent variable, indicated that sperm count and morphology are the most important seminal parameters for fertility assessment. Discrimination analysis revealed that sperm numbers and morphologic variables provide significant information for discriminating between fertile men and infertile men with normal or abnormal seminal characteristics. Sperm penetration and motility were not indicated as important factors. The present data suggest that, in unselected male patients seeking reproductive evaluation, the sperm penetration assay did not yield any additional information on the cause of infertility.

55 citations


Journal ArticleDOI
TL;DR: It is concluded that water resorption in the rat epididymis is responsive to aldosterone, and sperm concentrations were normal in animals receiving aldrenalectomy plus ald testosterone replacement.
Abstract: Experiments have been performed to determine if aldosterone is involved in the control of water reabsorption from the epididymal lumen in vivo. Micropuncture samples of lumen content were collected from the epididymides of control rats and those receiving aldrenalectomy, adrenalectomy + 25 micrograms aldosterone/day, 10 mg spironolactone/kg body weight/day, 10 mg spironolactone + 1 mg testosterone/kg body weight/day, 5 mg desoxycorticosterone acetate (DOCA)/day, 50 micrograms aldosterone/day, or 0.1 ml vehicle alone. The treatment period was three days. Seminal vesicles weights and testis weights were obtained. Sperm concentrations (SEM) in the caput, corpus, and cauda epididymidis of normal rats were 0.75 +/- 0.05, 1.24 +/- 0.13, and 1.99 +/- 0.15 x 10(9) sperm/ml, respectively. Both inhibition and removal of aldosterone caused significant reduction (P less than .01) of intraluminal sperm concentrations. Sham treatment had no effect. Sperm concentrations were normal in animals receiving aldrenalectomy plus aldosterone replacement. It is concluded that water resorption in the rat epididymis is responsive to aldosterone.

52 citations


Journal ArticleDOI
TL;DR: Six subjects who were evaluated by semen analysis and hormone profile at two-week intervals during ten-week basal, 20-week treatment, and post-treatment periods lasting at least ten weeks produced reversible oligozoospermia without attendant change in potency.
Abstract: We have previously shown that LHRH agonist [D-Trp Pro -Net]LHRH (LHRHA) results in reversible oligozoospermia when given to normal subjects for up to 10 weeks. A fall in plasma testosterone was accompanied by loss of libido and potency. The authors now report on 6 subjects who were evaluated by semen analysis and hormone profile at 2-week intervals during 10-week basal 20-week treatment and posttreatment periods lasting at least 10 weeks. Treatment consisted of LHRHA (50 mcg subcutaneously daily) and testosterone enanthate (100 mg intramuscularly every 2 weeks). Sperm density (mean basal 76.7 +or- 8.7x10 /ml) fell consistently in each subject to a mean nadir of 12.3 +or- 4.5x10 /ml (P /ml achieved when LHRHA was given alone. In each individual subject sperm density returned to his basal level after cessation of treatment. No consistent changes were seen in sperm motility or morphology or in semen volume. Libido and potency were maintained in all subjects. An additional 3 subjects received testosterone enanthate alone in identical dosage for 20 weeks. No change in sperm density was observed. In contrast to treatment with LHRHA alone combination treatment produces reversible oligozoospermia without attendant change in potency. (authors)

51 citations


Journal ArticleDOI
TL;DR: The inability of (+)gossypol to decrease male fertility is demonstrated and it is suggested that the activity of racemic gOSSypol may be due primarily to the presence of the (-) optical isomer.
Abstract: ( + )-Gossypol was isolated from the bark of Thespesia populnea and tested for its ability to inhibit the fertility of male hamsters. Male hamsters of proven fertility were treated orally for 54 days with 40 mg/kg of ( + )-gossypol, 40 mg/kg of racemic gossypol, or 5% gum acacia (vehicle control) and were mated with estrous female hamsters during the fourth and seventh weeks of treatment. Both the control and the (+ )-gossypol-treated animals exhibited normal fertility throughout the experiment. The racemic gossypol-treated animals were infertile when evaluated during both the fourth and seventh weeks of treatment. Morphologic examination of the testicular tissue could not explain the loss of fertility. These data demonstrate the inability of (+)gossypol to decrease male fertility and suggest that the activity of racemic gossypol may be due primarily to the presence of the (-) optical isomer.

46 citations


Journal ArticleDOI
TL;DR: It was concluded that phospholipase activity in BSA may contribute to capacitation and the acrosome reaction of guinea pig sperm in vitro by mimicking the action of native sperm phospholIPase.
Abstract: Phospholipase activity associated with guinea pig spermatozoa during capacitation and the acrosome reaction (AR) was determined by measuring the production of [14C] glycerylphosphorylcholine (GPC) from [14C] phosphatidylcholine (PTC) over a 2-hour period at the beginning and at the end of a 6-hour incubation. Spermatozoa converted 13 X more [14C] PTC to [14C] GPC during the last 2 hours of incubation than during the first 2 hours which corresponded to an increase in AR from 7% at 2 hours to 55% at 6 hours. In vitro studies also were performed to assess the role of phospholipase activity in bovine serum albumin (BSA) in inducing the AR of guinea pig spermatozoa. Spermatozoa, obtained by backflushing the cauda epididymidis, were incubated for up to 6 hours in Biggers, Whitten, and Whittinghams' (BWW) medium alone or containing various additives. These additives were: 0.1% BSA, phospholipase A2, p-bromophenacyl bromide, DMSO, 0.1% BSA plus p-bromophenacyl bromide, or 0.1% BSA plus DMSO. Each hour of incubation, samples were assessed for the percentage of AR sperm and sperm motility. The percentage of AR sperm after 6 hours in BWW supplemented with phospholipase A2, at the level of PTC hydrolyzing ability detected in BWW with 0.1% BSA, was similar to that of BWW and BSA. However, the percentage of AR sperm in BWW with BSA and the phospholipase inhibitor p-bromophenacyl bromide was significantly lower than that in BWW with BSA or phospholipase A2 and similar to that in BWW without BSA. Sperm motility was significantly less in incubations containing phospholipase A2, but lacking BSA. It was concluded that phospholipase activity in BSA may contribute to capacitation and the AR of guinea pig sperm in vitro by mimicking the action of native sperm phospholipase.

42 citations


Journal ArticleDOI
Zhuang Lz1, David M. Phillips1, Glen L. Gunsalus1, C W Bardin1, J P Mather1 
TL;DR: Of the somatic cell types in the testis, the Sertoli cells are most sensitive to gossypol, and there was a significant decrease in androgen binding protein (ABP) secretion by SERToli cells in the presence of gossYPol.
Abstract: In order to evaluate the direct effect of gossypol on testicular cells, we used primary cultures of rat Leydig and Sertoli cells. No alteration in Leydig cell survival, morphology, or testosterone production was seen during three days of culture with up to 3 microgram/ml gossypol. With higher concentrations (3 to 7 microgram/ml) of gossypol, there was a reduction in cell survival but no change in androgen secretion. In contrast, there was a marked change in Sertoli cell morphology after five days of gossypol treatment. Large vacuoles and electron dense granules appeared in the cytoplasm, but these effects were reversed within six days of removing gossypol from the medium. There was a significant decrease in androgen binding protein (ABP) secretion by Sertoli cells in the presence of gossypol. We also tested the effect of gossypol on the growth of three established cell lines. Two Sertoli-derived cell lines, TM4 and TR-ST, were more sensitive than a Leydig-derived cell line (TM3). These results suggest that, of the somatic cell types in the testis, the Sertoli cells are most sensitive to gossypol.

41 citations


Journal ArticleDOI
TL;DR: Data suggest a new theory, that increased pressure on the venous side of testicular capillary beds in subjects with varicocele may decrease blood flow and cause testicular damage.
Abstract: The blood flow rates and ability to remove an added fluid load were studied in the testes of monkeys with surgically induced varicoceles and in sham-operated control monkeys. These studies were conducted to verify a proposed mechanism by which varicocele may cause testicular and spermatogenic damage. Analyses of the data indicated that, at four months after induction, the testes of monkeys with varicocele were less able to remove a perfused fluid load and that, at five months, the blood flow was significantly less in the testes of these animals. The effects were most pronounced in the left testes of the monkeys with varicocele. Blood flow values calculated were: control, right--9.66 ml X 100 g-1 X min-1; control, left--10.42 ml X 100 g-1 X min-1; varicocele, right--7.73 ml X 100 g-1 X min-1; varicocele, left--5.04 ml X 100 g-1 X min-1. These data suggest a new theory, that increased pressure on the venous side of testicular capillary beds in subjects with varicocele may decrease blood flow and cause testicular damage.

Journal ArticleDOI
TL;DR: Purification of extracts of human semen by TRH affinity chromatography or gradient, reversed phase high pressure liquid chromatography revealed two major TRH immunoreactive peptides, one corresponding to the TRH-homologous peptide previously reported in rat and human prostate and one cochromatographing with synthetic TRH.
Abstract: High levels of thyrotropin releasing hormone (TRH) and TRH-homologous peptide, with the general structure pGlu-X-Pro-NH2 where X is either Histidine (His) or a neutral amino acid residue, have been identified in rat and human prostate (Pekary et al, 1980, 1981a, 1982c). Because of the secretory nature of prostatic tissue, it was considered likely that these peptides would be measurable in human semen. The mean (± SD) TRH immunoreactivity (TRH IR) in semen from 26 normal donors, aged 26 to 41 years, was found to be 12.2 ± 5.2 ng/ml. TRH and TRH-homologous peptide, which constitute most of the TRH IR of human semen, were undetectable in unconcentrated human serum. TRH IR levels in semen from azoospermic donors were in the normal range. The level of TRH IR in semen from vasectomized donors was significantly less (P < 0.01) than in semen from normal subjects. Evidence for at least two TRH-binding substances, which coextract and bind to added synthetic TRH, was obtained by exclusion and cation exchange chromatography. Purification of extracts of human semen by TRH affinity chromatography or gradient, reversed phase high pressure liquid chromatography (HPLC) revealed two major TRH immunoreactive peptides, one corresponding to the TRH-homologous peptide previously reported in rat and human prostate and one cochromatographing with synthetic TRH.

Journal ArticleDOI
TL;DR: In this article, the sperm Penetration Assay (SPA) results were treated with doxycycline and reevaluated by SPA and seminal fluid analysis and the improvement in SPA results was associated with a decrease in the number of leukocytes/ml in seminal fluid.
Abstract: Thirty-two men with abnormal (less than or equal to 11% penetration) Sperm Penetration (hamster ova) Assay (SPA) results were treated with doxycycline and reevaluated by SPA and seminal fluid analysis. Eighteen of the 32 infertile men with initially abnormal SPAs had normal SPAs (greater than or equal to 15% egg penetration) after treatment. None of 30 untreated infertile men with abnormal SPAs had a normal SPA on repeat testing (P less than 0.001). Improvement in SPA results was associated with a decrease in the number of leukocytes/ml in seminal fluid. Seven of the wives of the treated men conceived, while none of the wives of the untreated infertile men conceived during comparable intervals of time.


Journal ArticleDOI
TL;DR: The results indicate that the epididymal epithelium synthesizes and secretes glycoproteins, and that 1 to 2 hours are required for terminal glycosylation, intracellular transport, and release of the secretory product.
Abstract: The secretory activity of the middle segment of the mouse caput epididymidis was studied using 3H-fucose as a precursor to glycoprotein. Young adult male mice were injected with a concentrated solution of 3H-fucose interstitially, ie beneath the connective tissue capsule of the epididymis. Two animals were killed and prepared for light microscopic radioautography at each of six intervals between 10 minutes and 24 hours after injection. Silver grains were concentrated over the supranuclear Golgi region at 10 minutes and over the apical ends of the cells 30 minutes and 1 hour after injection. Quantitative analysis showed that luminal radioactivity increased greatly beginning with the 2-hour samples. The results indicate that the epididymal epithelium synthesizes and secretes glycoproteins, and that 1 to 2 hours are required for terminal glycosylation, intracellular transport, and release of the secretory product.

Journal ArticleDOI
TL;DR: Data show that isolated epithelial cells retain some differentiated structural features that characterize the intact cell and that enriched cultures of epithelial Cells can be maintained under conditions where fibroblast proliferation is inhibited.
Abstract: Epithelial cells isolated from the caput epididymidis of adult rats were placed in primary culture and examined daily for ten days for changes in external anatomy, reorganization of cytoskeletal components, maintenance of characteristic cytoplasmic features, and response to media formulated to minimize nonepithelial cell proliferation. Significant cell attachment to the substrate began after the first 24 hours of culture. After attachment, the cells underwent a progressive flattening and became closely applied to the substrate. This was accompanied by a redistribution of microvilli on the cell surface and a reorganization of cytoskeletal elements within the cell. After flattening, the cultured cells displayed an extensive array of 10-nm filaments which were associated with the desmosomes attaching adjacent cells. Immunofluorescence studies demonstrated that these were keratin-containing intermediate filaments and 2-D gel electrophoresis of intact cells and cell cytoskeletons revealed that a family of "keratin-like" polypeptides were major components of the cells. Epithelial cell attachment, morphology, and maintenance in the primary culture were unaffected by D-valine, cytosine arabinoside, or both; however, these agents, either individually or in combination, reduced significantly the number of cells incorporating 3H-thymidine. These data show that isolated epithelial cells retain some differentiated structural features that characterize the intact cell and that enriched cultures of epithelial cells can be maintained under conditions where fibroblast proliferation is inhibited.

Journal ArticleDOI
TL;DR: Leydig cells, purified on two sequential Percoll gradients to purities of 89 +/- 1%, were used to study the binding of 17-beta-hydroxy-17 alpha-methyl-estra-4,9,11-triene-3-one(3H-R1881), and the nuclear accumulation was androgen-specific.
Abstract: Leydig cells, purified on two sequential Percoll gradients to purities of 89 +/- 1%, were used to study the binding of 17-beta-hydroxy-17 alpha-methyl-estra-4,9,11-triene-3-one(3H-R1881). The accumulation of 3H-R1881 in the nuclear fraction of these cells was time- and temperature-dependent. Specific binding was saturable with an apparent Ka of 0.14 nM-1 and a single class of binding sites at a concentration of 721 fmol/mg DNA. A fraction of the bound radioactivity in the nuclear pellet could be extracted with 0.4 M KCl, and a portion of this extracted steroid was associated with macromolecular species. The nuclear accumulation was androgen-specific. These data are consistent with the presence of androgen receptors in rat Leydig cells.

Journal ArticleDOI
TL;DR: The comparison of these findings with those obtained previously with pituitary-grafted rats and with tumor-bearing rats indicates that the endocrine effects of hyperprolactinemia may vary depending on the rate and magnitude of the increase in peripheral PRL levels.
Abstract: Levels of plasma gonadotropins, prolactin (PRL), and testosterone, and the testicular concentration, total content, and affinity of hCG binding sites were measured in male spontaneously hypertensive rats (SHR) and in genetically matched normotensive (WKY) rats. Hypertensive rats had higher plasma PRL and FSH levels and lower plasma testosterone levels. The affinity of testicular hCG binding sites was similar in SHR and WKY rats, but SHR had considerably lower concentration and total content of hCG-binding sites than did WKY animals. The comparison of these findings with those obtained previously with pituitary-grafted rats and with tumor-bearing rats indicates that the endocrine effects of hyperprolactinemia may vary depending on the rate and magnitude of the increase in peripheral PRL levels. Species differences in the response to hyperprolactinemia and differences between SHR and other rat strains suggest that hormonal responses to PRL elevation depend also on the genetic characteristics of the animal.

Journal ArticleDOI
TL;DR: It is suggested that acrosin is not exclusively located on the inner acrosomal membrane and, thus, could participate in physiologic events other than zona penetration.
Abstract: Immunocytochemical techniques were employed to determine, at the ultrastructural level, the location of acrosin in porcine spermatozoa. Antisera to highly purified porcine acrosin was produced in rabbits. The (Fab')2 fragments of the immunoglobulins were purified and conjugated with horseradish peroxidase (HRP). Washed, formaldehyde-fixed spermatozoa were reacted with the labeled antiacrosin immunoglobulins, utilizing a direct staining technique. Electron microscopy revealed that the peroxidase reaction product of HRP-antiporcine acrosin was distributed evenly over the outer acrosomal membrane of spermatozoa with intact acrosomes. The labeled antibody was also distributed evenly over the inner acrosomal membrane of cells when the overlying acrosomal structures were absent. In some spermatozoa, labeling was noted throughout the acrosomal matrix. No significant labeling was observed in control specimens when spermatozoa were exposed to HRP-antiporcine acrosin immunoglobulins that had been adsorbed previously with excess purified acrosin or exposed to HRP-conjugated rabbit antiporcine immunoglobulins. This pattern of labeling is consistent with the hypothesis that acrosin may function as a zona lysin. The observation that the outer acrosomal membrane and acrosomal matrix are labeled suggests that acrosin is not exclusively located on the inner acrosomal membrane and, thus, could participate in physiologic events other than zona penetration.

Journal ArticleDOI
TL;DR: In this article, the effects of age and hemicastration on daily sperm production (DSP) were investigated in Sprague-Dawley rats with 6 to 10 months and 22 to 24 months.
Abstract: To investigate the effects of age and hemicastration on daily sperm production (DSP), testes from adult (6 to 10 months) and aged (22 to 24 months) Sprague-Dawley rats were evaluated before and four months after hemicastration. Testes were weighted, decapsulated, homogenized in a Waring Blender, and evaluated for the number of maturation-phase spermatids in the homogenates. DSP per testis was calculated by dividing the number of spermatids in the homogenates by the estimated life span of the spermatids (6.3 days). The same testicular traits, along with plasma LH and FSH levels, were determined in a second group of aged rats. Considering both testes in the first experiment, neither DSP per testis nor DSP per g parenchyma were significantly affected by age. However, the age testis interaction was significant (P < 0.01) in both parameters. DSP per g of the testis removed at hemicastration was slightly lower (P < 0.05) in the aged group, and DSP per g in the aged rat was enhanced (P < 0.05) in the hemicastrates. Unlike the adults, aged rats had enhanced (P < 0.01) DSP per testis in the remaining testis posthemicastration. In the second experiment, testicular weight, DSP per g, and DSP per testis were enhanced (P < 0.01) after hemicastration. Plasma FSH levels were significantly (P < 0.01) elevated within one week after hemicastration and remained elevated four months later. Plasma LH levels were not elevated within one week after hemicastration but were significantly (P < 0.01) elevated four months following hemicastration. We concluded that: 1. measured by DSP, no decline in spermatogenesis was found between 6 to 10 and 22 to 24 months of age; 2. aged rats were capable of hemicastration-induced testicular enlargement and enhancement of DSP, 3. rats in the adult group were not capable of these hemicastration-induced changes, 4. posthemicastration testicular enlargement was accompanied by an increased number of germ cells; 5. immediate and continued elevation of plasma FSH suggests its involvement in hemicastration-induced testicular enlargement of aged rats.

Journal ArticleDOI
TL;DR: The ability to fractionate the major polypeptide of the boar sperm PM into different and much smaller populations in large quantity and to characterize these proteins by electrophoresis provides a starting point for determining the functional significance and location of the major surface proteins of the boat sperm PM.
Abstract: High resolution, two-dimensional PAGE was used to characterize purified plasma membranes (PM) from boar spermatozoa. In addition to the abundance of polypeptides resolved, unusual features of PM discerned in gel patterns include a large number of basic polypeptides and the presence of at least three polypeptides with molecular weights in excess of 200 K daltons. In order to reduce these large numbers of polypeptides into smaller fractions, an important step in developing a surface map of these membranes, detergent-solubilized membranes were fractionated by the techniques of ion exchange chromatography and lectin affinity chromatography. PM polypeptides, solubilized in the nonionic detergents, were fractionated on DEAE Sephadex using phosphate ion gradients containing EDTA. Most of the major acidic polypeptides were eluted as a group at high phosphate ion concentrations. Membrane polypeptides can be separated into smaller populations by affinity chromatography on wheat germ agglutinin-agarose (WGA) or by Concanavalin A-agarose. Concanavalin A-agarose binds most PM glycoproteins, but partial purification of an important group of very basic proteins was obtained by repeated chromatography of polypeptides that failed to bind to this lectin. Since biologic activity (including immunologic potency) is often retained in the detergents used, the ability to fractionate the major polypeptide of the boar sperm PM into different and much smaller populations in large quantity and to characterize these proteins by electrophoresis provides a starting point for determining the functional significance and location of the major surface proteins of the boar sperm PM.

Journal ArticleDOI
TL;DR: Estradiol valerate had a marked impact on the elemental composition of luminal fluid only in the caput epididymidis--where sperm maturation is initiated--and a minor effect on that of cauda epididycumidal fluid--in which mature spermatozoa are stored.
Abstract: The maturation of spermatozoa in the epididymis is dependent upon the presence of androgens. This study examined the effects of androgen suppression by estradiol valerate on the elemental composition of the intraluminal fluids of the testis and epididymis. In the fluid from the caput epididymidis, the concentrations of sodium (106.1 +/- 3.4 to 182.8 +/- 16.9 mmol/1, P less than 0.01) and chloride (16.5 +/- 2.2 to 79.3 +/- 10.8 mmol/1, P less than 0.01) rose after treatment with estradiol valerate. By contrast, this treatment reduced the concentrations of phosphorus (63.7 +/- 1.6 to 47.8 +/- 3.2 mmol/1, P less than 0.01), sulfur (18.4 +/- 1.0 to 10.8 +/- 1.0 mmol/1, P less than 0.01), calcium (0.93 +/- 0.09 to 0.50 +/- 0.07 mmol/1, P less than 0.01), and magnesium (2.21 +/- 0.41 to 0.76 +/- 0.16 mmol/1, P less than 0.01). In the distal cauda epididymidis, the concentration of chloride rose after treatment with estradiol valerate (24.4 +/- 1.7 to 54.9 +/- 3.9 mmol/1, P less than 0.01), but the concentrations of the other measured elements (sodium, potassium, phosphorus, calcium, magnesium, and sulfur) were not altered by estrogen treatment. In rete testis fluid the concentration of phosphorus fell (2.00 +/- 0.30 to 0.67 +/- 0.12 mmol/1, P less than 0.01), while that of calcium rose (0.66 +/- 0.15 to 1.55 +/- 0.21 mmol/1, P less than 0.01). Estrogen treatment did not appear to affect the elemental composition of seminiferous tubular fluid or serum. Therefore, estradiol valerate had a marked impact on the elemental composition of luminal fluid only in the caput epididymidis--where sperm maturation is initiated--and a minor effect on that of cauda epididymidal fluid--in which mature spermatozoa are stored.

Journal ArticleDOI
TL;DR: The similarity in hormonal and tissue responses observed between dogs effectively immunized against LHRH and men with isolated gonadotropin deficiency suggests that the L HRH-immunized dog may provide a suitable experimental model for the study of patients with isolated Gonadotropins deficiency.
Abstract: Morphologic changes in the male reproductive system of mongrel dogs immunized against LHRH were quantitated using linear measurements and morphometric techniques at the light-microscopic level. Two experimental groups (5 nonimmunized control animals and 5 actively immunized animals) were killed 12 weeks after the primary immunization. No significant differences were observed between three immunized dogs having low LHRH antibody titers (immunized-unaffected) and the five nonimmunized control dogs. The two immunized dogs (affected) with the highest antibody titers against LHRH were characterized by atrophy and dedifferentiation of the testes, prostate, and excurrent ducts. The morphologic changes in the testes of these two dogs were striking and included an apparent arrest or significant reduction in the spermatogenic process, concurrent epithelial degeneration, and apparent diminution of Leydig cell mass. Drastic reductions in the size of the prostatic acini and epithelial cells, as well as loss of secretory granules, reflected depression of function and androgen production. Similarly, in the excurrent ducts decreases in the measured parameters and loss of regional cytoplasmic specialization denoted functional decrescence. This study demonstrates the regressive effects of LHRH immunoneutralization on the morphology of the reproductive system in the male dog and further supports the feasibility of this system as an animal model for the study of isolated gonadotropin deficiency.

Journal ArticleDOI
TL;DR: The results give evidence for a clear cut androgen-dependence in the appearance of the acid phosphatase present in semen, which therefore has been identified as secretory.
Abstract: Acid phosphatase was purified from human prostatic tissue and from seminal plasma Antisera to antigens from both sources were raised in rabbits These antisera have been used for immunohistochemical localization of the respective antigens in the prostates of neonatal, infantile, prepubertal, and adult individuals Immunoreactivity of the prostatic epithelium with the seminal fluid-derived antigen developed progressively in pubertal specimens with increasing age It was not present in fetal and infantile organs Antiserum prepared from human prostatic tissue-derived acid phosphatase gave a positive immunoreaction both with stroma and epithelium of the pre- and postpubertal glands The results give evidence for a clear cut androgen-dependence in the appearance of the acid phosphatase present in semen, which therefore has been identified as secretory The second antigen is nonsecretory, tissue-bound, nonandrogen dependent, and shares antigenic determinants with leukocyte-derived acid phosphatase

Journal Article
TL;DR: The degree of inhibition of spermatogenesis was insufficient to abolish fertility in the treated male rats, and the lack of effect on accessory organ weights are consistent with the maintenance of libido in these treated rats.
Abstract: A highly potent agonist of LHRH, [6-D-(2-naphthyl)-alanine]-LHRH, was administered chronically for 12 weeks to adult male rats by repetitive implantation of pellets, and its effects upon mating, fertility, and reproductive organ weights have been evaluated. Although significant declines in testicular (P less than 0.001) and epididymidal (P less than 0.001) weights were achieved, no effects on seminal vesicles, prostate, or pituitary weights were observed. After 12 weeks of continuous treatment, three of six agonist-treated rats were still successfully impregnating females. The decline in successful impregnation appeared to be related to the observed reduction in testicular spermatogenesis and in numbers of epididymal spermatozoa. The drug effects appeared reversible, as all six of the agonist-treated rats were fertile by the fifth week after cessation of treatment. Plasma levels of testosterone were markedly elevated immediately after implantation of each pellet and consistently, but not significantly, lowered during the inter-implantation periods. These observations, and the lack of effect on accessory organ weights, are consistent with the maintenance of libido in these treated rats. This is the second demonstration of a selective inhibition of spermatogenesis in the absence of a marked decline in gonadal steroidogenesis with this agent. As in the first demonstration using twice weekly injections, the degree of inhibition of spermatogenesis was insufficient to abolish fertility in the treated male rats.

Journal ArticleDOI
TL;DR: Results indicate that bilateral cryptorchidism severely impairs the secretion of inhibin and possibly other Sertoli cell functions which may account, at least partly, for the increase in circulating FSH levels and the arrest of spermatogenesis.
Abstract: The present study explored the effects of experimental bilateral cryptorchidism (of 21-, 28-, and 35-days duration) and orchidopexy (14 and 42 days) in the adult rat on the secretion of inhibin by cultures of isolated Sertoli cells. Changes in serum levels of gonadotropins, testis weight, and spermatogenesis also were assessed to verify the effectiveness of the surgical procedures. Cryptorchidy resulted in a progressive decline in testicular weight and a loss of germ cells, associated with increasing serum levels of FSH and LH. Inhibin secretion in vitro became nondetectable by 28 days after surgery. At 42 days after orchidopexy, spermatogenesis showed qualitative recovery, with a small increase in testes weight. Levels of LH in the circulation declined, but only to twice the intact control levels. However, inhibin secretion and serum FSH levels returned to nearly normal values. These results indicate that bilateral cryptorchidism severely impairs the secretion of inhibin and possibly other Sertoli cell functions which may account, at least partly, for the increase in circulating FSH levels and the arrest of spermatogenesis. The effects of cryptorchidism on these parameters can be reversed to a large degree by orchidopexy.

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TL;DR: A 22-year-old man with normal basal serum testosterone and luteinizing hormone (LH) levels but undetectable levels of serum FSH is studied, who demonstrated cryptorchidism, hypospadias, surgically repaired omphalocele, and bilateral hearing loss.
Abstract: Previous reports concerning isolated follicle stimulating hormone (FSH) deficiency and its possible pathogenesis have been conflicting. Both "normal" and "abnormal" FSH response to luteinizing hormone releasing hormone (LHRH) infusion have been described. We studied a 22-year-old man with normal basal serum testosterone and luteinizing hormone (LH) levels but undetectable levels of serum FSH. His serum LH titers showed one secretory spike during a 40-hour sampling at 20-minute intervals, whereas his serum FSH titers remained undetectable (less than 0.4 IU/l). Infusion of LHRH, 0.2 microgram/minute for 4 hours, induced the expected rise in the serum LH levels, but serum FSH levels remained low and only at one point reached 0.9 IU/l (normal adult male basal range 0.9-10.3 IU/l). The patient received LHRH, 100 micrograms/day, for three days. A second LHRH infusion, 0.2 microgram/minute for 4 hours, induced a normal rise in both the serum LH and FSH titers. The serum sex steroid binding globulin level was 10.3 ng DHT bound/ml (normal adult male level 8.0 +/- 0.3 ng DHT bound/ml). Presence of circulating auto-antibodies to the serum FSH was excluded by determining the binding of [125I] FSH with the patient's serum and comparing it with sera obtained from two normal male adult volunteers. Pituitary function tests were otherwise intact. Presence of a pituitary tumor was excluded by computerized axial tomography and x-ray studies of the pituitary fossa and normal visual fields. Clinically, the patient demonstrated cryptorchidism, hypospadias, surgically repaired omphalocele, and bilateral hearing loss.(ABSTRACT TRUNCATED AT 250 WORDS)

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TL;DR: In this paper, the authors used a test for nonprogressive motility based on the inability of a spermatazoon to leave a cube 200 micron on each side in less than 4 seconds.
Abstract: We have previously reported that epididymal spermatozoa from mice carrying a tw32 haplotype have a lower net velocity than do spermatozoa from congenic +/+ males. Using a test for nonprogressive motility based on the inability of a spermatazoon to leave a cube 200 micron on each side in less than 4 seconds, the frequency of nonprogressively motile sperm in these populations now has been determined to be above 90% after 2 to 5 hours of incubation in vitro. The nonprogressive motility never appears in sperm populations incubated in media with less than 0.1 mM Ca2+, although normal motility is well-maintained. Nonprogressive motility was also observed at low frequencies in uterine sperm populations, and at higher frequencies among uterotubal sperm from both +/+ and tw32/+ males. These observations suggest that nonprogressive motility may be a component of normal sperm function, either during sperm transport within the oviduct, or during fertilization.

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TL;DR: There were higher proportions of spermatozoa with abnormal morphology, total number of anomalies, and multiple anomalies in infertile men, both with and without varicocele, than in fertile men.
Abstract: This study was carried out to evaluate the morphology of spermatozoa in infertile men with and without varicocele. A series of 285 ejaculates were fully evaluated for seminal volume, sperm count, motility, and morphology, and classified into fertile (165 subjects), infertile without varicocele (93 subjects) and infertile with varicocele (27 subjects). Sperm morphology was classified by multiple entry criteria and recorded as normal, abnormal with head, midpiece, or tail single anomaly or abnormal with simultaneous multiple abnormalities. Semen volume was almost identical in the three groups. Among the infertile men, sperm count was lower in those having a varicocele, but conversely those with varicocele had a higher percentage of motile spermatozoa, higher percentage of spermatozoa with forward movement and higher sperm velocity. There were higher proportions of spermatozoa with abnormal morphology, total number of anomalies, and multiple anomalies in infertile men, both with and without varicocele, than in fertile men. The percentage of abnormal spermatozoa was higher in infertile men with varicocele than in those without varicocele. The pattern of sperm morphology differed between the infertile and the fertile group, and with the presence or absence of varicocele. In the presence of varicocele, only the incidence of elongated (tapered) forms was significantly increased.

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TL;DR: Results suggest that reduced protein kinase activity might contribute to the decreased testosterone synthesis of aged rats.
Abstract: The effects of aging on cyclic AMP (cAMP) dependent protein kinase activity of purified Leydig cells were evaluated. Purified Leydig cells from 20-day-, 40-day-, 60-day- and 30-month-old rats were incubated with or without hCG for 1 hour. At the end of incubation, tubes were centrifuged and supernatants were saved for cAMP and testosterone determinations, and the pellets were prepared for protein kinase assay. Basal cAMP levels of Leydig cells from 20-day- and 40-day-old rats were lower than those of 60-day- and 30-month-old rats. In response to hCG stimulation, Leydig cells of 40-day-old rats produced the highest amounts of cAMP, while 30-month-old rats had the lowest levels. Testosterone responses of 30-month-old rats were also significantly lower than those of 60-day-old mature rats. When cAMP-dependent protein kinase activities were measured, 30-month-old rats had the lowest total kinase activity. These results suggest that reduced protein kinase activity might contribute to the decreased testosterone synthesis of aged rats.