bluuebunny/crossref_metadata_2025 huggingface dataset:
Pituitary and Testicular Function in Spontaneously Hypertensive Rats
Authors: AMADOR A. , STEGER R. W. , BARTKE A. , JOHNS A. , HAYASHI R. H. , STALLINGS M. H.
Published: December 2015

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.
Improvement in the Sperm Penetration (Hamster Ova) Assay (SPA) Results After Doxycycline Treatment of Infertile Men
Authors: BERGER RICHARD E. , SMITH W. DIANNE , CRITCHLOW CATHY W. , STENCHEVER MORTON A. , MOORE DONALD E. , SPADONI LEON R. , HOLMES KING K.
Published: December 2015

Thirty‐two men with abnormal (< 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 (>15% egg penetration) after treatment. None of 30 untreated infertile men with abnormal SPAs had a normal SPA on repeat testing (P < 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.
The Effect of Exogenous Follicle Stimulating Hormone Treatment on Anterior Pituitary and Testicular Function in the Prepubertal Bull1
Authors: MEYERS STUART A. , SWANSON LLOYD V.
Published: December 2015

The effect of exogenous follicle stimulating hormone (FSH) on testicular function was studied in twenty 4‐month‐old Holstein bulls during two seasons (ten in late winter and ten in summer). Five mg FSH or saline were given subcutaneously at 12‐hour intervals (10 mg/day) for ten days, after which time the testes were removed, weighed, and incubated in vitro with 0,15, or 150 ng luteinizing hormone (LH) for 3 hours. FSH treatment resulted in significantly heavier testes (38%) in the summer (39.3 ± 1.6 vs 28.5 ± 1.5 g) but not in late winter. Epididymal weight was not affected. Across seasons, serum LH concentrations were higher (P = 0.06) in FSH‐treated bulls (2.39 ± 0.13 vs 1.80 ± 0.13 ng/ml); the largest increase occurred in the summer. FSH treatment did not change serum testosterone levels in late winter, whereas in the summer serum testosterone concentrations were threefold higher (0.90 ± 0.06 vs 0.29 ± 0.02 ng/ml; P < 0.05) in the FSH‐treated bulls. In addition, serum testosterone concentrations increased with time of FSH treatment. FSH treatment stimulated a 2.4‐fold increase (P < 0.05) in testosterone content of the testes across seasons and a threefold increase (P < 0.05) in testosterone synthesis in vitro in the summer. There was no dose effect of LH on in vitro testosterone synthesis. We conclude that administration of exogenous FSH to intact prepubertal bulls is capable of altering testicular function, but that seasonal influences can modify this function.
Human Semen Contains Thyrotropin Releasing Hormone (TRH), a TRH‐homologous Peptide, and TRH‐binding Substances
Authors: PEKARY A. EUGENE , HERSHMAN JEROME M. , FRIEDMAN STANLEY
Published: December 2015

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.
Testicular Blood Flow and Fluid Dynamics in Monkeys with Surgically Induced Varicoceles
Authors: HARRISON RICHARD M. , LEWIS RONALD W. , ROBERTS JAMES A.
Published: December 2015

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 mat, 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 × 100 g−1 × min−1; control, left—10.42 ml × 100 g−1 × min−1; varicocele, right—7.73 ml × 100 g−1 × min−1; varicocele, left—5.04 ml × 100 g−1 × 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.
Data on Virilization and Erotosexual Behavior in Male Hypogonadotropic Hypopituitarism During Gonadotropin and Androgen Treatment
Authors: CLOPPER RICHARD R. , MAZUR TOM , MacGILUVRAY MARGARET H. , PETERSON RALPH E. , VOORHESS MARY L.
Published: December 2015

The goal of this investigation was to assess whether or not gonadotropin therapy enhanced the degree of virilization and psychosexual behavior of men with hypogonadotropic hypopituitarism. Pre‐ and post‐gonadotropin assessments of virilization in four men indicated that gonadotropin therapy was associated with dramatic improvements in the degree of virilization that each man previously obtained on androgen only. Retrospective interview data on erotosexual behavior indicated improved erotosexual function on gonadotropin as compared to the prior androgen treatment. These data suggest that complete virilization in these men was partially gonadotropin dependent. Whether or not the behavioral benefits reported by these men represented a direct or synergistic effect of gonadotropin in the expression of erotosexual behavior or an indirect effect of improved virilization can not be answered by these data.
Pulsatile Secretion of Gonadotropins and Prolactin in Male Marathon Runners Relation to the Endogenous Opiate System
Authors: ROGOL ALAN D. , VELDHUIS JOHANNES D. , WILLIAMS FREDERICK A. , JOHNSON MICHAEL L.
Published: December 2015

We tested the hypothesis that sustained, strenuous physical training alters the neuroendocrine regulation of pulsatile gonadotropin and/or prolactin secretion in men. Blood was sampled at 20‐minute intervals over 8 flours in five endurance‐trained men after a 10–15 mile run in the middle of the active training season, and in 11 nonendurance trained normal controls. In these two groups, basal patterns of physiologically pulsatile secretion of LH, FSH, and prolactin (PRL) were not significantly different in relation to the following parameters: mean serum concentration of each of the three tiormones (N = 25 samples); areas under the hormone concentration vs. time curves; fractional, incremental, ind absolute pulse amplitudes; and pulse frequency, or periodicity. To test for enhanced suppressive effects of endogenous opiates in trained male marathon runners, subjects were administered the potent opiate‐receptor antagonist, naltrexone (1 mg/kg). This antagonist significantly stimulated pulsatile LH secretion by increasing mean serum LH values from 10.94 to 13.58 mIU/ml (P = 0.007); area under the LH concentration vs. time curve increased from 5370 to 6510 mIU/ml ± 3 hours (P = 0.05) and, pulse frequency rose from 2.8 to 4.9 pulses/8 hours (P = 0.006). Naltrexone also enhanced pulse frequency of FSH secretion from 3.4 to 5.4 pulses/8 hours (P = 0.009), but did not alter serum prolactin concentrations. None of these responses differed significantly from those in normal sedentary controls. We concluded that 1) detailed parameters of physiologically pulsatile LH and FSH secretion, even in men pf endurance training, are not distinguishable from those of normal sedentary controls; 2) secretion of PRL basally and in response to opiate‐receptor blockade is not significantly perturbed; and 3) LH release in male athletes appears to be under tonic inhibitory control by endogenous opiate systems, but the inhibitory influence of these opiates is not markedly exerted.
Periodic Exposure to a Brief Light Signal Stimulates Neuroendocrine‐Gonadal Activity in Golden Hamsters
Authors: EARNEST DAVID J. , TUREK FRED W.
Published: December 2015

The photoperiodic effects of a periodic light pulse on neuroendocrine‐gonadal activity in the male golden hamster was examined using a night interruption paradigm. Adult male hamsters that had been housed 3 to 5 per cage on LD 14:10 were placed either in individual cages, each equipped with a running wheel, or maintained in communal housing conditions, without access to a running wheel. Animals were then transferred to either LD 6:18 or to a LD 6:18 light cycle with a periodic 10‐second night interruption (8 hours after lights off) occurring once every two, four, or seven days. As expected, exposure to LD 6:18 for 11 weeks induced complete regression of the testes and seminal vesicles, accompanied by low serum levels of LH and FSH, in both individually and communally‐housed animals. However, in individually‐housed animals receiving a 10‐second night interruption every other day on LD 6:18, paired testis and seminal vesicle weights, as well as serum gonadotropin levels, were maintained at values comparable to those normally observed in hamsters exposed to photostimulatory long days. Furthermore, the presentation of a periodic 10‐second night interruption once every four or seven days to individually‐housed animals with access to a running wheel was sufficient to partially or totally block the inhibitory effects of short days on neuroendocrine‐gonadal activity. Communally‐housed hamsters receiving a 10‐second light pulse once every two, four, or seven days also exhibited paired testis and seminal vesicle weights, as well as serum gonadotropin levels, that were consistently higher than the values obtained for animals exposed only to LD 6:18. However, the efficacy of each night interruption paradigm in maintaining testicular and seminal vesicle weight in communally‐housed animals was considerably less than that observed for animals with access to a running wheel.

These results demonstrate that the periodic presentation of a brief photic stimulus once every two, four, or seven days during the dark phase of an inhibitory light cycle can influence the neuroendocrine events underlying the photoperiodic regulation of gonadal function. This experimental approach, whereby a discrete stimulus is periodically administered at a specific arcadian time to induce a photoperiodic response, should provide a valuable model for examining the physiologic mechanisms by which light regulates the neuroendocrine‐gonadal axis in mammals.
The Effect of Prepubertal Spermatic Cord Torsion on Subsequent Fertility in Rats
Authors: COSENTINO M. JAMES , RABINOWTTZ RONALD , VALVO JOHN R. , COCKETT ABRAHAM T. K.
Published: December 2015

This study was undertaken to determine the effects of various durations of testicular torsion in prepubertal rats on their subsequent fertility, and to determine whether these effects could be altered by removal of the torsioned testis. Sixty rats (35 days old) were subjected to 720° unilateral spermatic cord torsion for 0,1, 3,5,9, or 12 hours. The torsioned testis was then either detorsioned or removed. At 65 days of age each male was housed with two females for three weeks. Rats undergoing detorsion of the spermatic cord demonstrated a linear decrease in fertility with respect to the duration of torsion (r = −0.904). However, all of the animals undergoing unilateral torsion with subsequent orchiectomy were fertile, regardless of the duration of torsion. In addition, the percentage of females impregnated, the number of embryos produced, and the mean embryo size decreased with increasing intervals of torsion (r = −0.834 to r = −0.979); the sharpest decline occurred between 5 and 9 hours of torsion. All of these parameters were significantly lower (P < 0.001 to P < 0.05) in the detorsioned group as compared to the orchiectomized group. There was a decrease in seminiferous tubule diameter in the contralateral testis with respect to the duration of torsion (P < 0.01). These data indicate that unilateral spermatic cord torsion in young rats significantly reduced their subsequent fertility with respect to duration of the torsion, and that this detrimental effect may be minimized if the damaged testis is removed rather than untwisted and replaced into the scrotum.
Bromocriptine in the Management of Infertile Men after Surgery of Prolactin Secreting Adenomas
Authors: MANCINI ANTONIO , GUITELMAN ABRAHAM , LEVALLE OSCAR , APARICIO NESTOR , ASZENMIL GUSTAVO
Published: December 2015

This study evaluated bromocriptine treatment in nine patients with prolactin secreting adenomas who continued to have elevated circulating levels of prolactin after surgery, and who were interested in improving their sperm counts. These patients were brought into the study 3.2 ± 1.8 years (mean ± SD) after surgery. All of them presented with high circulating levels of prolactin, and eight of the patients had oligozoospermia (range 0–10 × 106 spermatozoa/ml). LH and testosterone levels were low in seven patients, and eight patients had low FSH values. All patients were treated for 90 days with 7.5 mg/day of bromocriptine. After treatment, prolactin levels decreased significantly in all patients, while sperm counts increased significantly in five of them. Testosterone levels increased in four subjects. Bromocriptine therefore seems useful in the management of this type of patient because of the observed decline in prolactin levels and the increase in sperm counts. Possible mechanisms involved in this action are discussed.
The Effects of Ethylene Glycol Monomethyl Ether on Testicular Histology in F344 Rats
Authors: CHAPIN ROBERT E. , DUTTON SANDRA L. , ROSS MONICA D. , SUMRELL BILLIE M. , LAMB JAMES C.
Published: December 2015

Ethylene glycol monomethyl ether (EGME) has been found to produce testicular atrophy in experimental rodents. The studies that follow were designed to determine the testicular cell type(s) most susceptible to EGME administration. For histologic studies, F344 rats were gavaged with 150 mg/kg/day of EGME 5 days per week, and serially sacrificed. In sections from perfusion‐fixed tissue, necrotic changes were observed in some meiotic and premeiotic spermatocytes 24 hours after a single dose. Also, nuclear condensation was seen in occasional early pachytene spermatocytes. These effects were magnified after two doses; there were more necrotic pachytene and meiotic spermatocytes than necrotic stage I pachytene spermatocytes. By day 4, testes from all treated animals were affected; there was a pronounced maturation‐depletion effect, seen as the absence of round spermatids from tubules in stages I to III. These effects continued to develop at days 7 and 10, leaving only Sertoli cells, spermatogonia, and late stage spermatids populating the epithelium. Other animals were treated similarly, but subject to efferent duct ligation 16 hours prior to sacrifice. Fluid production, as judged by weight gain in the testes after efferent duct ligation, was unaffected by EGME treatment. Analysis of the fluid collected at the rete testis indicated that there was no treatment‐related change in the relative amounts of androgen binding protein. The data indicate that the spermatocyte is the primary target cell for the histologic effects of EGME in the testis of F344 rats.
Physiopathologic Aspects of Leydig Cell Function in Varicocele Patients
Authors: ANDÒ S. , GIACCHETTO C. , COLPI G. , BERALDI E. , PANNO M. L. , LOMBARDI A. , SPOSATO AND G.
Published: December 2015

Leydig cell function was studied in 108 varicocele (V) patients with a mean age of 30.9 years, and a control group (C) of 46 men with a mean age of 30 years. Plasma gonadotropin levels were determined before and after GNRH stimulation. Testosterone (T), 17‐OH‐progesterone (17‐OH‐P), dihydrotestosterone (DHT) and estradiol (E2) were also assayed.

Mean plasma T levels were significantly decreased in varicocele patients (V = 416 ± 12.9, n = 106; C = 487 ± 19.9, n = 40; P < 0.01), while the basal 17‐OH‐P/T ratio was significantly increased (V = 0.38 ± 0.02, n = 56; C = 0.28 ± 0.02, n = 40; 0.02 > P > 0.01) and remained higher after hCG stimulation (P < 0.01). No significant differences in mean sex steroid levels were observed when comparing varicocele patients with normal sperm counts (VN) and those who had oligozoospermia (VO). There was a significant negative linear correlation between age and 17‐OH‐P (n = 56; r = −0.47; P < 0.01) and T values (n = 106; r = 0.27; P < 0.01) in varicocele patients, which contrasted with the absence of any significant correlation with age in the controls. These data suggest that the duration of idiopathic varicocele influences testicular hormone secretion.
Human Chorionic Gonadotropin Treatment of Male Sexual Inadequacy in Patients Affected by Chronic Renal Failure
Authors: CANALE DOMENICO , BARSANTINI STEFANO , MINERVINI RICCARDO , FIORENTINI LUCIO , BARSOTTI GIULIANO , MENCHINI‐FABRIS G. FABRIZIO
Published: December 2015

Twenty patients with sexual inadequacies secondary to different degrees of chronic renal failure were investigated. The patients were divided into two groups: group A—ten patients treated with a conservative diet therapy; group B—ten patients on hemodialysis. All the patients under investigation were administered 2000 IU of hCG every other day for 120 days after a 30‐day period without treatment. Andrologic, biometric, hematologic, and sexual data were evaluated before and after hormone treatment. Sexual parameters were collected through self‐evaluation by the patients and their partners.

Our results agree with Van Kammen et al (1975) and show an improvement of the andrologic and biometric data in patients from group A. Furthermore, eight patients form group A and three patients from group B reported a remarkable improvement in their sexual responses.
Evaluation of Penile Circulation with the Doppler Technique
Authors: ANNONI FEDERICO , LANIA CATERINA , GRASSO MARCO , RONCHI FRANCO , PRICOLO VICTOR E.
Published: December 2015

The Doppler ultrasound technique was used to examine 72 patients complaining of nonsustained erection. In 27 subjects an evident hemodynamic alteration in penile circulation was found. In 12 of these patients, associated alterations of the hypogastric artery were present, while in the remaining 15, pure arterial lesions, due to trauma or to La Peyronie's disease, were detected.
Alloplastic Bridge Spermatocele
Authors: PUPPO PAOLO , BELGRANO EMANUELE , GABOARDI FRANCO , TROMBETTA CARLO
Published: December 2015

A new type of artificial spermatocele is proposed. A Gore‐tex cylinder is sutured to the cauda of the epididymis, making a bridge between the two testes. The advantages of this type of spermatocele are: 1) use of a single puncture; 2) easy execution of the puncture because the bridge stabilizes and holds the testicles in place, 3) greater quantity of aspirate and therefore better conservation of spermatozoa; 4) better quality of aspirate; and 5) relative ease of the surgical procedure.
Linear and Nonlinear Mouse Sperm Motility Patterns: A Quantitative Classification
Authors: TESSLER STEVEN , OLDS‐CLARKE PATRICIA
Published: December 2015

A quantitative method is described for distinguishing two types of mouse sperm motility using a new parameter, the Linear Index. One hundred spermatozoa from C57BL/6‐+/+ and 100 spermatozoa from C57BL/6‐tw32/+ mice were used to evaluate this parameter. Spermatozoa were tracked by videomicrography, and the following calculated: net displacement velocity, Vn; curvilinear velocity of the sperm head, Vc; the “average” velocity, Va (a 5‐point moving average of the track); the progressiveness ratio, (Vn/Vc); the curvilinear progressiveness ratio, (Va/Vc); and a linear index, (Vn/Va). Frequency histograms of linear index values were bimodal for both groups and each had a clear antimode at 0.5, separating two discrete subpopulations of spermatozoa. Within each group, “linear” (>0.5) and “nonlinear” (<0.5) tracks differed in all characteristics; between groups, linear spermatozoa were similar and nonlinear spermatozoa differed only in the curvilinear velocity of the sperm head.
Effects of White Blood Cells on the In Vitro Penetration of Zona‐free Hamster Eggs by Human Spermatozoa
Authors: MARUYAMA DONALD K. , HALE RALPH W. , ROGERS B. JANE
Published: December 2015

The presence of white blood cells in semen has been associated with male infertility. Previous studies indicate that pyospermia occurs in conjunction with decreases in sperm motility, number of normal sperm forms, and penetration rates in the zona‐free hamster egg sperm penetration assay. We have evaluated the relationship of seminal white blood cells and sperm function, as reflected in the zona‐free hamster egg penetration assay, and have investigated the possible mode of action of the white cells. Egg penetration rates decreased when white blood cells from fertile or potentially fertile donors were added to their sperm suspensions prior to preincubation and at insemination in the in vitro assay. Zona‐free hamster egg penetration assay results were also inhibited when the supernatant from white blood cells incubated in Biggers, Whitten, and Whittingham (BWW) medium overnight were introduced to sperm‐oocyte suspensions at insemination. Conversely, egg penetration rates were enhanced in samples from hypofertile individuals when white blood cell concentrations in the semen or WBC/sperm ratios were reduced, either by physical removal or as a result of antibiotic therapy. The physical presence of leukocytes, and possibly, the extracellular release of lysosomal enzymes may be responsible for the inhibitory effects in vitro. Although the mechanism(s) by which white blood cells interfere with the fertilizing capacity of spermatozoa are not clear, it is quite obvious that their presence in the in vitro environment is undesirable and can mask an individual's actual fertilizing potential.
Alterations of Testicular Function Induced by Hyperprolactinemia in the Rat
Authors: KATOVICH MICHAEL J. , CAMERON DON F. , MURRAY FREDERICK T. , GUNSALUS G. L.
Published: December 2015

The effects of hyperprolactinemia on testicular structure and pituitary‐gonadal function were investigated in male rats. Hyperprolactinemia was induced in the Wistar‐Furth rat by implantation of tissue fragments of a prolactin‐secreting MtTW15 pituitary adenoma. The MtTW15 tissue was maintained in one animal group for 27 days (group A) and in another group for 37 days (group B). Appropriate age‐matched controls were utilized in each study. Serum prolactin was significantly elevated (P < 0.001) in both groups of MtTW15‐bearing rats compared with their controls. The degree of hyperprolactinemia was more severe in rats of group B (2842 ± 546 ng/ml) than in rats of group A (367 ± 38 ng/ml). Accessory sex organ weights in group B rats were significantly lower than in controls, but were apparenty unaffected in group A rats. Hyperprolactinemia induced definite but variable testicular alterations in both animal groups that presented as seminiferous epithelial disorganization, germ cell exfoliation, increased tubule wall thickness, and abnormal Leydig cell lipid content. Electron microscopy revealed structural disruption of Sertoli‐germ cell junctional complexes and apical Sertoli cell cytoplasmic degeneration. The hyperprolactinemic rat exhibited significant reductions in serum luteinizing hormone (LH), testosterone (T), and androgen binding protein (ABP) when compared with controls. Eighty‐six days following surgical removal of the MtTW15 tissue in a subgroup of group B rats, serum levels of prolactin and LH returned to normal, as did weights of accessory sex organs and testicular morphology. These results indicate that exposure to the MtTW15 adenoma and its later removal in the rat provides a workable model for studying the effects of hyperprolactinemia on testicular structure and function, and for identifying events involved in the subsequent recovery of spermatogenic disruption.
Decreased Fertility and Motility of Spermatozoa from Rats Immunized with a Prealbumin Epididymal‐specific Glycoprotein
Authors: FOURNIER‐DELPECH S. , COUROT M. , DUBOIS M. P.
Published: December 2015

This study investigated the effects on the progressive motility, zona‐binding capacity, and fertility of spermatozoa from the cauda epididymidis of adult male rats that were actively immunized against an acidic glycoprotein secreted by the epididymis. The percentage of motile spermatozoa was less than 5% in nine of ten rats that received the epididymal antigen, and 40 to 50% in eight of the 10 control rats. In animals immunized against the antigen, there was a dramatic decrease, but not a complete suppression, in the capacity of epididymal spermatozoa to bind the zona pellucida as compared with the non‐immunized controls. Fertility was decreased two weeks after the end of the treatment, but partial restoration of fertility was observed 6 months later.
Culture of Ciliated and Nonciliated Cells from Rat Ductuli Efferentes
Authors: BYERS STEPHEN W. , MUSTO NEAL A. , DYM MARTIN
Published: December 2015

The isolation and culture of ciliated and nonciliated cells from rat ductuli efferentes is described. Fragments of epithelium obtained after two collagenase digestions attached to plastic and to extracellular matrix and could be maintained in culture for at least 2 weeks. Ciliary beating in cells grown on epididymal extracellular matrix‐coated plastic could be observed for up to 7 days in culture. Although cells maintained on this substrate retained organelles characteristic of cells in vivo, they assumed a flattened, squamous appearance. In contrast, cells growing on the surface of permeable supports impregnated with extracellular matrix were polarized and exhibited a cuboidal/columnar appearance. Androgen binding protein conjugated to colloidal gold was taken up by these cells via coated pits and was found sequentially in uncoated endosomes, multivesicular bodies and lysosomes.
Inhibition of LDH‐X by Gossypol Optical Isomers
Authors: KIM IN CHULL , WALLER DONALD P. , FONG HARRY H. S.
Published: December 2015

Racemic gossypol is an effective male antifertility agent in several mammalian species. However, (+)‐gossypol is not an effective male antifertility agent in the rat or the hamster. Previous studies have demonstrated the ability of racemic gossypol to inhibit the testis‐spedfic LDH‐X enzyme derived from various mammalian species and have suggested LDH‐X as the potential site of gossypol antifertility action. In the present study, the effects of racemic gossypol and the (+) and (—) optical isomers of gossypol on LDH‐X derived from rat and hamster testicular cytosol are compared to determine if there is any correlation between the in vitro inhibition of the LDH‐X enzyme and in vivo antifertility effects. Both optical isomers of gossypol as well as racemic gossypol inhibit rat and hamster testicular cytosolic LDH‐X activity. Inhibition of hamster testicular cytosolic LDH‐X activity by (–)‐gossypol was less than by either racemic gossypol or (+)‐gossypol. Based on the previous reports of racemic gossypol inhibition of LDH‐X, therefore, it cannot be simply concluded that LDH‐X is the specific site of antifertility action of gossypol since, in the present study, (+)‐gossypol, which is not an effective male antifertility agent, also inhibited rat and hamster testicular cytosolic LDH‐X.
Epididymal Fluid Macromolecules Do Not Act as Auto‐ or Alloantigens
Authors: ESPONDA P. , BEDFORD J.M.
Published: December 2015

Because epididymal secretory glycoproteins form important functional associations with maturing spermatozoa, the possibility has been explored that epididymal antigens might be useful in contraceptive vaccine development. Male and female rabbits, hamsters, and rats were immunized for several weeks with epididymal fluid, initially either with complete Freund's adjuvant, or conjugated with glutaraldehyde. Some female rabbits were also immunized using albumin‐free rabbit epididymal fluid. The sera of all the immunized animals were then examined for autogenous, allogeneic, or [in guinea pigs also] xenogeneic immune responses, using immunodiffusion and ELISA techniques. In each instance, xenogeneic immunization produced a marked antibody response. However, no animals developed any detectable antibody reactive with epididymal fluids following auto‐ or alloimmunization (ie with fluid from the same species), and they remained fertile even when immunized males were mated with immunized females. Furthermore, when female rabbits were immunized with ejaculated rabbit spermatozoa, they produced antibodies reactive only with the sperm homogenate, but not with any epididymal fluid component. These results indicate that macromolecules secreted by the epididymis, including those that associate with spermatozoa, do not act as auto‐ or alloantigens and, at present, would seem to have no immediate promise for contraceptive vaccine development in males or females.
Possible Significance of Transferrin Levels in Seminal Plasma of Fertile and Infertile Men
Authors: FORESTA CARLO , MANONI FABIO , BUSINARO VITTORIO , DONADEL CARLA , INDINO MARIANGELA , SCANDELLARI CESARE
Published: December 2015

Human seminal plasma contains large amounts of transferrin, which is a protein secreted mostly by Sertoli cells. It has been suggested that the concentration of transferrin may serve as a possible clinical marker of Sertoli cell function. Therefore the concentration of this protein in human seminal plasma from fertile and infertile men has been evaluated in order to find a relationship between transferrin concentrations and human semen parameters and plasma FSH levels. Findings show that seminal transferrin in subjects with oligozoospermia or azoospermia is significantly lower than in controls, and that it is strongly related to sperm count. Results also indicate that transferrin secretion can be impaired when plasma FSH levels are still normal, suggesting that seminal transferrin is an early and specific marker of Sertoli cell function. These results, however, do not clarify whether impairment of transferrin secretion by Sertoli cells is due to an organic dysfunction or to an organic secretory alteration.
Increased Plasma and Pituitary Prolactin Concentrations in Adult Male Rats with Selective Elevation of FSH Levels May Be Explained by Reduced Testosterone and Increased Estradiol Production
Authors: SHI YI‐FAN , PATTERSON AMY PATE , SHERINS RICHARD J.
Published: December 2015

The roles of testosterone and estradiol in regulating prolactin concentrations were studied in acutely castrated adult male rats receiving subcutaneous Silastic® implants of the sex steroids. Testosterone was administered in increasing doses, from subphysiologic to intact levels, both alone and in combination with a small, single dose of estradiol. The study was designed to assess whether a change in the relative rates of sex steroid production could account for an increase in PRL release in the absence of other testicular factors. At very low levels of plasma testosterone, FSH and LH levels were indistinguishable from castrate controls. As plasma testosterone concentration increased, both plasma FSH and LH levels were suppressed progressively to intact levels. When a subphysiologic dose of testosterone was coadministered with a small dose of estradiol, the combined effects produced a midcastrate level of FSH but maintained a normal level of LH similar to the selective increase in FSH concentration observed in men with germinal aplasia. Although PRL levels were indistinguishable in intact and castrate controls, testosterone replacement by capsule increased prolactin in a dose‐related manner so that, at the physiologic level of testosterone, prolactin was elevated two‐fold (P< 0.01), similar to the level achieved with estradiol replacement alone. Pituitary prolactin levels also increased with increasing doses of testosterone but values remained within the range measured in intact controls. When estradiol was coadministered with testosterone, the combination produced different effects depending on the testosterone dose. With subphysiologic testosterone replacement, when plasma FSH was selectively increased, plasma PRL was elevated four‐fold (P< 0.01) compared with intact controls and two‐fold compared with rats receiving estradiol alone (P < 0.01). A smaller increase in pituitary PRL content was noted similar to the level achieved with estradiol replacement alone. As the testosterone dose increased toward midphysiologic values, plasma and pituitary prolactin levels fell progressively, but remained elevated above intact levels (P< 0.05). It was concluded that testosterone and estradiol both modulate prolactin secretion in the absence of other testicular factors, but the greatest effect is seen when the testosterone‐estradiol ratio is reduced, producing a selective increase in FSH concentration. These results are similar to the increased prolactin release observed in men with testicular dysfunction associated with selectively elevated FSH concentrations. The precise mechanism of these changes of prolactin release remain to be elucidated.
Lack of Correlation Between Fertility and Sperm Numbers in Male Rats Treated with Histrelin, a Potent LHRH Agonist
Authors: FOLDESY ROBIN G. , VANDERHOOF MARY M. , HAHN AND DO WON
Published: December 2015

Adult male rats were treated daily for up to 8 weeks with histrelin, [(ImBzl)‐D‐ His6,Pro9‐NEt]LHRH, to study the antifertility effects of this LHRH agonist. Although serum testosterone concentrations and testicular sperm numbers were significantly decreased by weeks 2 and 4 respectively, a reduction in fertility, as judged by the mean number of fetuses per mated female, was not observed until the sixth week of treatment. Furthermore, the number of spermatozoa in the cauda epididymidis of treated rats did not decrease below initial control levels at any time during the study and full fertility returned within 4 weeks after histrelin treatment was stopped. Thus, the lack of correlation between fertility and testicular and epididymal sperm numbers suggests that the antifertility effects of LHRH agonists are not due solely to reduced sperm numbers, but also result from androgen deficiency.
The Presence of a Motility Inhibitor Within Spermatozoa May Explain the Poor Sperm Motility of Some Infertile Men
Authors: de LAMIRANDE EVE , SHERINS RICHARD J. , GAGNON CLAUDE
Published: December 2015

The presence of motility inhibitors in seminal plasma and within spermatozoa from control and infertile men with poor sperm motility was investigated using demembranated reactivated human spermatozoa. No difference was found in the inhibitory capacities in seminal plasma of patients with poor sperm motility (< 50%) when compared with that of fertile controls with motility above 50%. No correlation was observed between inhibitory capacity and sperm motility. However, when extracts of spermatozoa from these patients were tested for the presence of inhibitor, it was observed that three of nine patients had an inhibitor in their sperm extract. By contrast, all sperm extracts from fertile control subjects were devoid of inhibitor. It was concluded that the presence of a motility inhibitor in seminal plasma does not explain the poor sperm motility observed in patients. The presence of a motility inhibitor within spermatozoa, however, may represent an important factor in the etiology of the poor sperm motility observed in some patients.
Repair of Experimental Varicoceles in the Rat Long‐term Effects on Testicular Blood Flow and Temperature and Cauda Epididymidal Sperm Concentration and Motility
Authors: HURT G. SAMUEL , HOWARDS STUART S. , TURNER TERRY T.
Published: December 2015

The effects of varicocele and varicocele repair on testicular blood flow, temperature, sperm counts, and sperm motility were assessed in adult male rats. The duration of the experimental varicocele and the varicocele repair were three and two times as long, respectively, as that studied previously. Varicoceles were created by partial ligation of the left renal vein and repairs were accomplished by high ligation of the left spermatic vein. Testicular blood flow was determined by using the radiolabeled microsphere technique. Testicular temperature was taken via needle probe thermometer. Sperm samples were obtained by micropuncture of the cauda epididymidis, and were counted on a hemacytometer and observed for motility under the light microscope. Varicoceles were studied 100 days after their creation. Repairs were performed on varicoceles that had lasted 100 days and the animals were studied 60 days after repair. Mean testicular blood flow (ml/100 g tissue/min) was significantly increased (P < 0.05) in animals with varicocele (left testis (LT) = 42.2 ± 1.1, right testis (RT) = 39.1 ± 1.2) when compared with normal controls (LT = 29.3 ± 1.6, RT = 29.6 ± 1.7), animals with varicocele repair (LT = 30.7 ± 1.3, RT = 30.0 ± 1.6), or sham‐operated animals (LT = 29.7 ± 1.4, RT = 31.1 ± 1.4). Testicular vs. intra‐abdominal temperature differences were significantly less (P < 0.05) for varicocele animals (ΔLT = 2.1 ± 0.1 C, ΔRT = 2.5 ± 0.1 C) as compared with normal controls (ΔLT 4.0 ± 0.5 C, ΔRT 4.1 ± 0.4 C), animals with varicocele repair (ΔLT = 4.0 ± 0.3 C, ΔRT = 4.1 ± 0.3 C), or shams (ΔLT ± 4.2 ± 0.1 C, ΔRT 4.4 ± 0.2 C). Sperm counts and motility also were reduced significantly (P < 0.05) by experimental varicocele and approached normal values following varicocele repair.
Antimitotic Drugs (Cyclophosphamide and Vinblastine) in the Male Rat Deaths and Behavioral Abnormalities in the Offspring
Authors: AUROUX MAURICE R. , DULIOUST EMMANUEL M. , NAWAR NABIL Y. , YACOUB SAMEH G.
Published: December 2015

The spermatogenesis and the offspring of male rats treated either with cyclophosphamide alone, or with both cyclophosphamide and vinblastine were investigated. The offspring were evaluated for the mean number of pups per litter, sex ratio, the frequency of apparent external malformations and, within the first 4 months of life, growth and mortality. When they reached adulthood and were between 12 and 16 weeks of age, the offspring were also examined for spontaneous activity and learning capacity.

Treatment with cyclophosphamide or cyclophosphamide and vinblastine resulted in a decrease in the number of both primary spermatocytes and spermatids; the effect, however, lasted longer for the combined drug regimen. At birth, the animals sired by the treated males did not show any apparent malformations. However, compared with the control population 1) the mortality rate of the offspring was significantly higher within the first 40 days of life; 2) at adult age, the proportion of animals that failed in the learning ability test was significantly increased and those that did succeed showed impaired learning capacity. The difference, however, was significant only in the male offspring. Finally, the offspring's spontaneous activity was significantly decreased. No difference was found in mortality or behavior between the animals born of the cyclophosphamide or cyclophosphamide plus vinblastine‐treated males.

The behavioral disorders shown in the adult offspring confirm the existence of a long‐term risk of paternal origin. This risk, essentially functional and independent of any morphologic pathology, should be taken into account in the context of environmental genotoxicity.
Testicular Hemodynamic Changes After the Surgical Creation of a Varicocele in the Rat Intravital Microscopic Observations
Authors: NAGLER HARRIS M. , LIZZA ELI F. , HOUSE STEVEN D. , TOMASHEFSKY PHILIP , LIPOWSKY HERBERT H.
Published: December 2015

The present study investigated the effect of a surgically induced varicocele on the dynamics of testicular blood flow. The surface vasculature of the normal and the varicocele‐affected testis was examined utilizing intravital epi‐illumination microscopy. Application of this technique to the study of the varicocele is new. Blood flow characteristics in surface veins were studied as the surface temperature of the testis was varied. Periodic, reproducible stoppages in blood flow, determined by direct observation of the red blood cells, were seen in seven of eight sham animals at the lower temperatures. These stoppages were abolished and blood flow increased at higher temperatures; stoppages reappeared at lower temperatures. The periodic stoppages were present in only one of eight rats with a proven varicocele (P < 0.025) at any temperature studied. This loss of blood flow regulation may be the result of a loss of testicular arteriolar tone and may explain the increase in testicular blood flow and temperature elevation observed in association with a varicocele. These findings may provide new insights into the pathophysiology of the varicocele and highlight the need to study the microvascular sequelae of this vascular abnormality.
Spontaneous Lipid Peroxidation and Production of Hydrogen Peroxide and Superoxide in Human Spermatozoa Superoxide Dismutase as Major Enzyme Protectant Against Oxygen Toxicity
Authors: ALVAREZ JUAN G. , TOUCHSTONE JOSEPH C. , BLASCO LUIS , STOREY BAYARD T.
Published: December 2015

Spontaneous lipid peroxidation in washed human spermatozoa was induced by aerobic incubation at 32 C and measured by malonaldehyde production; loss of motility during the incubation was determined simultaneously. Malonaldehyde production at the point of complete loss of motility, defined as the lipoperoxidative lethal end‐point (LLE), was 0.10 ± 0.03 nmol/108 cells (X̄ ± SD, n = 40), and was independent of the time to complete loss of motility. Human spermatozoa produced both H2O2 and O2−. during aerobic incubation. Inhibition of superoxide dismutase in these cells with KCN showed that all the H2O2 production is due to action of the dismutase. The superoxide dismutase activity of individual human sperm samples varied between 1 and 10 U/108 cells, variations between samples from a single donor being nearly as great as those between different donors. The time to complete motility loss (tL) showed equal variation of 1 to 10 hours among samples. The rate of spontaneous lipid peroxidation, calculated as LLE/tL, for a given sperm sample and the superoxide dismutase activity of the same sample, determined prior to aerobic incubation, gave a good linear correlation (r = 0.97). Glutathione reductase, glutathione peroxidase, and glutathione were found to be present in human spermatozoa, but showed little variation among samples. These results suggest that superoxide dismutase plays the major role in protecting human spermatozoa against lipid peroxidation. In addition, the superoxide dismutase activity of a fresh sperm sample appears to be a good predictor of the lifetime (up to the complete loss of motility) of that particular sample, and so may prove useful in semen analysis.
Uptake and Release of Ascorbic Acid by Rat Leydig Cells In Vitro
Authors: MOGER WILLIAM H.
Published: December 2015

The mechanism by which ascorbic acid is transported into rat Leydig cells in vitro was investigated. Transport of [14C]ascorbic acid was temperature‐dependent and inhibited in the absence of sodium in the incubation medium. The uptake of [14C]ascorbic acid was saturable (apparent Km ∼ 33 μM, Vmax ∼ 20 pmol/106 cells/min) and steriospecific. Uptake was not affected by removing glucose from the incubation medium, indicating that no significant uptake was occurring via the glucose transport system. Transport of [14C]ascorbic acid into Leydig cells was not affected by testosterone concentrations up to 7 μM or by LH at a concentration that causes maximum androgen secretion. Leydig cells preloaded with [14C]ascorbic acid rapidly released radioactivity into the incubation medium, a process that was unaffected by the presence of LH. The results suggest that ascorbic acid uptake by Leydig cells is by an electroneutral, sodium‐dependent co‐transport mechanism as in some, but not all, other types of cells.
Alkaline Phosphatase Histochemistry Discriminates Peritubular Cells in Primary Rat Testicular Cell Culture
Authors: CHAPIN ROBERT E. , PHELPS JERRY L. , MILLER BRUCE E. , GRAY TIM J. B.
Published: December 2015

Histochemical demonstration of alkaline phosphatase activity appears to be useful in identifying rat peritubular cells in primary testicular cell culture. In both frozen sections of rat testis and Mirsky's fixed, methacrylateembedded rat testis, the reaction product localized primarily in peritubular cells, vascular endothelium and occasionally in interstitial cells, with much smaller amounts of reaction product associated with elongating spermatids in the germinal epithelium. Occasional late‐stage tubules (X‐XIV) showed weak reactivity in the epithelium, associated with spermatocytes or Sertoli cells. Ultrastructurally, Gomori‐method reaction product was localized to peritubular cells, lymphatics, and spermatogonia in stage VII; no staining was found consistently in Sertoli cells. In isolated cell preparations enriched for Sertoli and germ cells, 1 to 8% of the cells demonstrated alkaline phosphatase activity, while greater than 50% of the cells stained positive for alkaline phosphatase activity in peritubular‐enriched fractions. The histochemical demonstration of alkaline phosphatase activity can be useful for identifying peritubular cells in primary cultures of testicular cells.
Comparison Between Proteinases of Human Seminal Plasma and of Sperm Origin
Authors: HUME MICHAEL E. , SIEGEL MARK S. , POLAKOSKI KENNETH L.
Published: December 2015

A comparison of the alkaline proteinase activity of human seminal plasma, the seminal non‐gamete cellular material and spermatozoa was made by gelatin‐sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (gelatin‐SDS‐PAGE) zymography. Several major (molecular weights = > 56,000) and minor (35,000 to 44,000) bands of proteinase activity were seen in the seminal plasma samples from nonvasectomized and vasectomized, healthy donors. Similar activity profiles were observed in the nongamete cellular material of vasectomized donor ejaculates. The major proteinase activity in sperm extracts was in the 47,000 to 55,000 (proacrosin‐acrosin) and 34,000 to 37,000 (sperminogen‐spermin) molecular weight ranges. These results suggest that the proacrosin‐acrosin and sperminogen‐spermin systems are of sperm origin and that there are considerable amounts of larger molecular weight trypsin‐like enzymes in the soluble and nongamete cellular material of human seminal plasma.