bluuebunny/crossref_metadata_2025 huggingface dataset:
The Neglected Laboratory Test The Semen Analysis
Authors: CHONG AUGUSTO P. , WALTERS CLIFFORD A. , WEINRIEB SHELLEY A.
Published: December 2015

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.
Human Sperm Penetration in Bovine Cervical Mucus Clinical Studies I
Authors: BORGHI MARIO R. , ASCH RICARDO H.
Published: December 2015

Penetration of spermatozoa in bovine cervical mucus (BCM) from two groups of men was investigated and correlated with the different parameters of spermograms from these same subjects. Sperm penetration and motility were statistically greater in men of proven fertility than in men from infertile couples in which a female cause of infertility had been ruled out. The percentage of cases with sperm penetration ≥15 mm was significantly greater in volunteers of proven fertility than in the group of men from infertile couples (100% and 33%, respectively). A marked correlation was observed between sperm penetration and sperm count (p < 0.0001) and between sperm penetration and motility (p < 0.0001). In a follow‐up of the group of men from infertile couples, it was found that all those who impregnated their wives within 18 months after the study presented sperm penetration ≥15 mm. It is suggested that BCM is an appropriate medium to determine sperm penetration in the investigation of infertile couples.
Organ Culture of Rat Caput Epididymal Tubules in a Perifusion Chamber
Authors: KLINEFELTER GARY R. , HAMILTON DAVID W.
Published: December 2015

We have been able to culture caput epididymal tubules from rats by modifying an organ culture system (Orgebin‐Crist et al, 1980) which was used to culture rabbit corpus epididymal tubule segments. The morphology of the epithelium was consistently good throughout seven days in culture, although sloughed epithelium was commonly seen during the first 24 hours. Evidence of this sloughing was much less frequent thereafter. Throughout seven days of culture, autoradiography of SDS‐PAGE of luminal fluid obtained from tubules cultured in medium containing 14C‐L‐leucine showed incorporation into bands identical to those stained by Coomassie Blue. Rat epididymal α‐lactalbumin was consistently localized on the luminal surface of the epithelium and the middle piece of the spermatozoa. Spermatozoa appeared to have normal morphology throughout the first three days in culture; thereafter, decapitated spermatozoa were frequently seen. Caput spermatozoa only quiver in place prior to culture, but after three days in culture, 53% of the spermatozoa from distal caput tubules are progressively motile upon dilution in a balanced salt solution. Since the transit time for spermatozoa in the caput epididymidis of the rat is approximately three days, it should be possible with this culture system to study maturational events involving interactions between spermatozoa and the epididymal epithelium.
A Prospective Study of the Relationship Between Semen Quality and Fertility in Cases of Unexplained Infertility
Authors: AITKEN R. JOHN , BEST FIONA S. M. , WARNER PAMELA , TEMPLETON ALAN
Published: December 2015

The male partners of 68 couples exhibiting 5.1 ± 0.3 (SEM) years of unexplained infertility were assessed using the conventional criteria of semen quality, the movement characteristics of the spermatozoa and the outcome of the zona‐free hamster egg penetration test. After a follow‐up period of 2.3 ± 0.06 (SEM) years, 25 (37%) of these patients were found to have initiated a pregnancy, thereby permitting an analysis of those aspects of semen quality which most accurately predicted their subsequent fertility. A multivariate discriminant analysis revealed that the conventional semen profile, per se, was not of significant value in discriminating the incidence of pregnancy. However, significant discrimination (P = 0.0173) was obtained when the postcapacitation movement characteristics of the spermatozoa were incorporated into the analysis. The accuracy of this prognosis was further increased if either the duration of infertility or the outcome of the zona‐free hamster egg penetration test was taken into consideration. Overall classifications of fertility were then 76.3% and 76.5% accurate, respectively. These results suggest that in vitro assessments of human sperm function are of significant value in evaluating male fertility.
Monolateral Tubulovasostomy: Report of Four Cases
Authors: BELGRANO E. , PUPPO P. , TROMBETTA C. , PITTALUGA P.
Published: December 2015

After conventional vasoepididymostony, the epididymal lumen may become occluded or obstructed, resulting in recurring azoospermia. Recently, two‐layer direct anastomosis has been advocated on the basis of experimental models of tubulovasostomy performed in rats. Monolateral tubulovasostomy was performed in four cases under 16–40 times magnification. Nylon or 11/0 Vycril® was used as suture material for the internal layer of the anastomosis, and 10/0 Ethilon® was used for the external layer. After three and six months, semen samples were evaluated for sperm count and motility. The presence of spermatozoa in all the samples confirmed 100% patency of the duct. Three patients had a number of spermatozoa compatible with fertility, and one pregnancy occurred.
Bovine Sperm Adenylate Cyclase: Inhibition by Adenosine and Adenosine Analogs
Authors: BROWN MEREDITH A. , CASILLAS EDMUND R.
Published: December 2015

Since both adenosine and cAMP affect sperm motility and metabolism, the authors have studied the mode of interaction of adenosine with the adenylate cyclase activity in either the presence or the absence of 15 mM caffeine. In general, structural analogs of adenosine containing alterations of the purine structure produce little or no inhibition, whereas analogs containing alterations of the ribose structure are effective inhibitors. Cyclic AMP and caffeine also inhibit the enzyme activity. The kinetics of the adenosine inhibition most closely resemble the linear noncompetitive type. Adenosine is a more effective inhibitor of adenylate cyclase activity in the presence of Mn2+ rather than Mg2+, and is more effective at higher metal ion concentrations. These observations are consistent with a P‐site interaction of adenosine with adenylate cyclase.
Secretion of Androgen Binding Protein by Sertoli Cells Is Influenced by Contact with Germ Cells
Authors: GALDIERI M. , MONACO L. , STEFANINI M.
Published: December 2015

Sertoli cells were cultured alone or with germ cells to evaluate the effect of the association with germ cells on the secretory activity of Sertoli cells. Secretion of androgen‐binding protein, which is specifically secreted by Sertoli cells, was measured under several experimental conditions. The following experimental models were utilized: 1) cultures of explants of seminiferous epithelium from prepubertal animals in which germ cells adherent to Sertoli cells are present (Sertoli cell enriched cultures); 2) monolayers formed only by Sertoli cells, obtained by removing germ cells from Sertoli cell enriched cultures, and 3) cocultures of Sertoli cell only cultures and germ cell populations at defined stages of differentiation. The results obtained indicated that FSH‐induced ABP secretion was greatly reduced in Sertoli cell only cultures as compared to enriched Sertoli cell cultures, and that this difference was stable throughout the first eight days of culture. In addition, cocultures of Sertoli cell only cultures with germ cells induced an increase of ABP when cocultured germ cells were at differentiation stages, such as pachytene spermatocytes, which are able to recognize and firmly adhere to the Sertoli cell monolayers. Cocultures with round spermatids, which do not adhere to Sertoli cells, did not increase the amount of FSH‐induced ABP production. The addition of nongerminal cells such as lymphocytes and fibroblasts were also not effective in stimulating ABP secretion. Surface interaction between Sertoli cells and cocultured germ cells seemed to be necessary for this FSH‐induced ABP production. When pachytene spermatocyte adhesion to Sertoli cells was prevented by the presence of an agar layer stratified above the monolayer, no increase in the FSH‐induced ABP secretion was observed. These data suggest that the FSH responsiveness is influenced by surface interactions with germ cells at defined stages of their differentiation. The possible mechanisms involved in the observed phenomenon are discussed.
Specific Inhibition of the Testicular Mitochondrial Respiratory Chain In Vitro by Gossypol
Authors: KIM INCHULL , WALLER DONALD P.
Published: December 2015

Optically inactive gossypol is an effective male antifertility agent in several mammalian species, while optically active (+)‐gossypol has no antifertility effect in the rat and hamster. Recently, it was suggested that the mitochondria of spermatogenic cells may be a subcellular target of gossypol. We are reporting the effects of optically inactive gossypol and (+)‐gossypol on the respiratory chain of mitochondria isolated from the testes and liver of rats and hamsters. The mitochondria were incubated with the test compounds and difference spectra were recorded. Complete inhibition of the testicular mitochondrial respiratory chain was observed at a concentration of approximately 75 μM. In contrast, no inhibition of the liver mitochondrial respiratory chain was observed with the test compounds at concentrations as high as 300 μM. These results demonstrate selective inhibition of the testicular mitochondrial respiratory chain by gossypol isomers.
Mechanism of Action of Gonadotropin‐releasing Hormone‐stimulated Leydig Cell Steroidogenesis.
Authors: LIN TU
Published: December 2015

The present study was designed to elucidate mechanisms responsible for gonadotropin‐releasing hormone (GnRH)‐stimulated testosterone formation. Purified Leydig cells from adult Sprague—Dawley rats were incubated with varying concentrations of GnRH agonist (des‐Gly10, (D‐Ala6) GnRH N‐ethylamide), hCG, 8‐bromo cAMP or pregnenolone; testosterone, cAMP, cyclic GMP (cGMP) and cAMP‐dependent protein kinase activity were measured after various time periods. Basal testosterone levels were 2.54 ± 0.13 ng/105 cells, increasing to 3.18 ± 0.14, 4.32 ± 0.08, and 4.63 ± 0.12 ng within 1 hour after the addition of 10−9, 10−8, and 10−7 M GnRH agonist, respectively. After a 3‐hour incubation a 10−7 M dose of GnRH agonist increased testosterone production four‐fold above control. GnRH agonist potentiated hCG‐stimulated testosterone formation, but had no significant effects on cGMP levels and cAMP‐dependent protein kinase activity. Cyclic AMP levels in the incubation medium increased slightly. GnRH agonist also enhanced 8‐bromo‐cAMP and pregnenolone‐induced testosterone formation. Furthermore, GnRH agonist increased testosterone formation both in the absence and presence of phosphodiesterase inhibitor. These results suggest that the major effect of GnRH agonist is probably beyond the cAMP step. When purified Leydig cells were incubated in a calcium‐free medium, the stimulatory effects of GnRH agonist on testosterone formation were completely abolished, but could be restored by the addition of calcium to the incubation medium. GnRH agonist‐induced testosterone formation was also blocked by the addition of nifedipine (a calcium channel blocking agent, 0.1 to 10 μg/ml). Finally, GnRH antagonist in a concentration of 10 μg/ml completely inhibited GnRH agonist‐stimulated testosterone formation. In conclusion: GnRH agonist stimulated Leydig cell testosterone formation in short‐term incubations. The stimulatory effect is calcium dependent and not mediated by cyclic nucleotides.
Gonadotropic Therapy in Men with Hypogonadotropic Hypogonadism
Authors: GATTUCCIO FRANCESCO , D'ALIA ONOFRIO , BARTOLO GIUSEPPE LO , ORLANDO GAETANO , JANNÌ ALBERTO , IZZO PIERLUIGI , CANALE DOMENICO , MENCHINI‐FABRIS G. FABRIZIO
Published: December 2015

Many therapeutic regimens, differing in dose and duration, have been employed in the treatment of hypogonadotropic hypogonadism. This study presents the results from a regimen of hCG plus hMG plus testosterone combined treatment in 15 patients affected by selective hypogonadotropic hypogonadism. The patients were homogeneous in age (16–23 years), testicular size, aspermia, absence of prior hormonal treatment, and normal karyotype. They were administered 5000 IU of hCG plus 75 IU of hMG twice a week for two months and a single dose of depo‐testosterone (250 mg) in the third month. This therapeutic plan was carried out continuously for eight three‐month cycles (24 months). Sexual maturation and seminal parameters were checked every six months. All the patients improved in testicular size (from 4.6 ± 1.55 ml to 13 ± 3.5 ml) and secondary sexual characteristics (penis, scrotum, pubic hair), but facial and axillary hair in most patients remained abnormal. After 24 months, all of them had ejaculations, but only two were normo‐spermic, and seven were still azoospermic. In patients with hypogonadotropic hypogonadism treated with gonadotropins there were better results in achieving sexual maturation than in improving seminal parameters. We believe that this differential response is due to multiple factors rather than a single factor.
Testicular LH Receptors During Aging in Fisher 344 Rats
Authors: AMADOR ARMANDO , STEGER RICHARD W. , BARTKE ANDRZEJ , JOHNS ANTHONY , SILER‐KHODR THERESA M. , PARKER C. RICHARD , SHEPHERD ALEXANDER M.
Published: December 2015

Levels of serum LH, prolactin, testosterone, progesterone and 17‐OH progesterone and the testicular concentration and total content of LH receptors were measured in 4‐, 11‐, 18‐, and 27‐month‐old Fisher 344 rats. All 27‐month‐old rats had Leydig cell tumors. At first, testicular LH receptor levels decreased with age, but with the appearance of the testicular tumors, these levels increased dramatically. Serum prolactin levels fluctuated with age, but were significantly decreased in 27‐month‐old rats, as were serum LH levels. Serum testosterone levels decreased steadily with age, while the testosterone‐LH receptor ratio remained constant until the appearance of the testicular tumors, after which the ratio decreased precipitously. Serum progesterone levels remained constant throughout the life of Fisher 344 rats until the appearance of testicular tumors, when they increased dramatically. Serum 17‐OH progesterone levels were increased significantly at 11 and 27 months as compared to four months of age, but levels at 18 months were similar to those seen in the 4‐month‐old animals. Therefore, in aged Fisher 344 rats with spontaneous Leydig cell tumors, there is an alteration in the testicular testosterone synthesizing pathway at a step after progesterone.
Changes in Serum Levels of LH, FSH, Prolactin, Testosterone, and Cortisol Associated with Season and Mating in Male Pygmy Goats
Authors: HOWLAND BORDEN E. , SANFORD LEE M. , PALMER W. MARTYN
Published: December 2015

Four male pygmy goats were used in a study designed to determine the effects of season on serum hormone (luteinizing hormone, follicle stimulating hormone, prolactin, testosterone, and cortisol) levels, testis size and libido, and the effects of mating on serum hormone profiles. Seasonal peaks were observed for prolactin in July, luteinizing hormone and follicle stimulating hormone in September, and testosterone in October. Luteinizing hormone peak frequency was greatest in September and was increased by mating activity in the months immediately preceding the breeding season. Scrotal circumference did not vary with season and libido showed no consistent seasonal pattern. Mating appeared to raise all hormone levels except during the months when these hormones were seasonally elevated. When episodic releases of luteinizing hormone occurred, they were associated with subsequent rises in serum testosterone levels. On some mating days, when episodic releases of luteinizing hormone were absent, changes in testosterone levels were highly correlated with changes in cortisol levels. It was concluded that both season and mating influence reproductive hormone levels in male pygmy goats.
Receptor‐mediated Endocytosis of Alpha2‐macroglobulin by Principal Cells in the Proximal Caput Epididymidis In Vivo
Authors: DJAKIEW DANIEL , BYERS STEPHEN W. , LEWIS DOROTHY M. , DYM MARTIN
Published: December 2015

Micropuncture techniques were used to study receptor‐mediated endocytosis of α2‐macroglobulin bound to colloidal gold (α2M‐gold) by principal cells in the proximal caput epididymidis of control and efferent duct‐ligated rats. The pathway of receptor‐mediated endocytosis of α2‐macroglobulin‐gold in vivo was similar to that which occurs in vitro. Alpha2‐macroglobulin‐gold was taken up and internalized in coated pits and coated vesicles and was localized sequentially in uncoated vesicles (endosomes), tubular‐vesicular structures, multivesicular bodies, and lysosomes. However, a 100‐fold excess of α2‐macroglobulin did not displace the uptake of α2‐macroglobulin‐gold in principal cells from control rats. In contrast, uptake of α2‐macroglobulin‐gold by principal cells from efferent duct‐ligated rats was six‐fold greater than in control rats, and could be displaced to control levels by a 100‐fold excess of α2‐macroglobulin. It is suggested that the inability of a 100‐fold excess of α2‐macroglobulin to displace uptake of α2‐macroglobulin‐gold in control rats was due to the normal saturation of apparent α2‐macroglobulin receptors on principal cells. The effect of efferent duct ligation was to remove the high levels of endogenous α2‐macroglobulin, which depleted the receptors of α2‐macroglobulin, thereby allowing a higher uptake of α2‐macroglobulin‐gold in the efferent duct‐ligated rats.
Localization of Cellular Retinol‐binding Protein and Cellular Retinoic Acid‐binding Protein in the Rat Testis and Epididymis
Authors: PORTER STEVEN B. , ONG DAVID E. , CHYTIL FRANK , ORGEBIN‐CRIST MARIE‐CLAIRE
Published: December 2015

The distribution of cellular retinol‐binding protein (CRBP) and cellular retinoic acid‐binding protein (CRABP) in rat testis and epididymis was examined by the peroxidase‐antiperoxidase immunolocalization technique. In the testis, cellular retinol‐binding protein was localized exclusively in the Sertoli cells. Staining varied with the stages of the seminiferous epithelium cycle and was maximal prior to the maturation divisions. Cellular retinoic acid‐binding protein was localized exclusively in the germinal cells in the adluminal compartment. The results suggest that retinoic acid may be the retinoid form used by the germinal cells, and that Sertoli cells may use the cellular retinol‐binding protein to transfer retinol from the basal to the adluminal compartment. In the epididymis, cellular retinol‐binding protein was localized in the cytoplasm and stereocilia of the principal cells in the proximal caput epididymidis, while cellular retinoic acid‐binding protein was localized in the spermatozoa and the stereocilia of the principal cells throughout the epididymis and in the epithelial cells of the distal vas deferens. Sperm staining intensity decreased from the initial segment to the cauda. The presence of high levels of cellular retinol‐binding protein in the epithelial cells and high levels of cellular retinoic acid‐binding protein in the spermatozoa of the caput epididymidis, known to be involved in the synthesis and secretion of factors necessary for sperm maturation, suggests that vitamin A may have a role in this process.
The Origin of Some Acid Hydrolases of the Fluid of the Rat Cauda Epididymidis
Authors: MAYORGA LUIS S. , BERTINI FRANCISCO
Published: December 2015

The activity of N‐acetyl‐β‐D‐glucosaminidase (NAG, 60.1 units/mg protein) and of acid phosphatase (57.7 units/mg protein) in fluid from the cauda epididymidis formed without any contribution from the testis (fluid obtained from a perfused and isolated cauda epididymidis or from an epididymis whose corresponding efferent ducts had been ligated for 40 days) was significantly higher than the activity of these enzymes in normal fluid (39.6 and 41.2 units/mg protein, respectively). Arylsulphatase activity of the locally formed fluid (11.2 units/mg protein) was lower than that of normal fluid (74.1 units/mg protein). The rete testis fluid was relatively rich in arylsulphatase since the ratio of arylsulphatase to acid phosphatase activity was 17 times higher in this fluid than in locally formed fluid. It is concluded that the activities of NAG and acid phosphatase in normal fluid from the epididymis originate in the epididymal tissue, while most of the arylsulphatase activity comes from the testis.
A Comparative Analysis of cAMP‐dependent Protein Kinase Regulatory Subunits in Sea Urchin and Rat Spermatozoa
Authors: KHATOON S. , HALEY B.E. , ATHERTON R.W.
Published: December 2015

8‐Azido cAMP photoaffinity labeling of cAMP‐dependent protein kinase regulatory subunits (R1 = 49 K; R2 = 55 K) was done on spermatozoa from species lacking, and species containing an epididymis. Spermatozoa from sea urchin and trout contained only R1, while rat caudaepididymal spermatozoa contained both R1 and R2 subunits. This was established by the Mr value of the 8‐azido cAMP photolabeled moieties, and a biochemical analysis based on the known differences of protein‐nucleotide interactions of Type I and II cAMP‐dependent protein kinases. Sea urchin and trout sperm Ri subunits were similar to mammalian sperm Ri subunits in co‐migration on SDS‐polyacrylamide gels and in both saturation and specificity of nucleotide binding. Calcium enhanced photoprobe binding to rat R1 and R2 subunits and to sea urchin Ri subunit without revealing a sea urchin R2 subunit. Likewise, phosphodiesterase incubation of sea urchin and trout spermatozoa prior to photolabeling did not reveal R2 subunits. These data suggest that the cAMP regulation of sperm physiology may require Ri subunit in species both with and without an epididymis. Further taxonomic study is necessary to determine whether evolutionary acquisition of the epididymis and internal fertilization may have created unique environments favoring the addition of sperm R2 regulatory subunits of cAMP‐dependent protein kinase.
Androgen Receptor Levels and Androgen Contents in the Prostate Lobes of Intact and Testosterone‐treated Noble Rats
Authors: HO SHUK‐MEI , DAMASSA DAVID , KWAN PAUL W.L. , SETO HENRY S.K. , LEAV IRWIN
Published: December 2015

Plasma testosterone (T) levels were correlated with androgen receptors, tissue content of T, and 5α‐dihydrotestosterone (DHT) in the three anatomically‐discrete prostate lobes of intact and castrated Noble (Nb) rats bearing T‐filled silastic capsules. Differences in androgen receptor content and tissue androgen levels were observed among the three prostatic lobes of intact Nb rats. Total (cytosolic and nuclear) androgen receptor levels were highest in the ventral prostate followed by the dorsolateral and anterior prostate lobes. In the ventral and anterior prostate, androgen receptors were found to be equally distributed between cytosols and nuclear extracts, whereas in the dorsolateral prostate, androgen receptors were predominantly nuclear (cytosolic: nuclear = 1:5). The ventral prostate had the highest total androgen content and DHT was the major tissue androgen in all three lobes. The ratio of tissue DHT:T varied among the lobes; the highest value was observed in the dorsolateral prostate. The higher proportions of nuclear androgen receptor, as well as the elevated tissue DHT:T found in the dorsolateral prostate compared to other lobes, suggest that differences in the androgen activation process may exist between the dorsolateral prostate and other prostatic lobes.

Despite lower plasma and tissue T levels, the DHT content, weight and cytodifferentiation in all lobes of T‐treated castrated rats closely approximated the situation found in intact animals. Total androgen receptor levels were, however, elevated in all prostatic lobes of T‐treated, castrated rats as compared to intact controls. These increases were primarily attributed to the augmented levels of androgen receptor in the nuclear extracts of the three prostate lobes. Exposure of the prostate to a constant level of T, produced by silastic implantation, might be responsible for the higher total androgen receptor levels and enhanced nuclear androgen receptor retention found in the prostates of T‐treated, castrated rats.
Flow Cytometric DNA Analysis of Defined Stages of Rat Seminiferous Epithelial Cycle During In Vitro Differentiation
Authors: TOPPARI JORMA , EEROLA ERKKI , PARVINEN MARTTI
Published: December 2015

The stages of the rat seminiferous epithelial cycle have been isolated for flow cytometric analysis of DNA and for culture, using transillumination‐assisted microdissection. Precise stages have been identified by phase contrast microscopy of live cell squashes from adjacent segments. Each stage of the cycle showed a characteristic flow cytometric pattern with haploid (1C), diploid (2C) and tetraploid (4C) peaks. Stages I to VIII of the cycle showed an additional hypofluorescent (0.25–0.70C) peak due to a reduced dye‐binding capacity of maturation phase‐spermatids at steps 15 through 19. The appearance of the hypofluorescent haploid peak coincided with the second nucleoprotein transition at step 15 of spermiogenesis and the homogeneous condensation of the chromatin seen in electron microscopy. As a concomitant of the formation of disulphide bonds during epididymal maturation, the fluorescence intensity decreased further to reach a relative value of 0.07C in the cauda epididymidis. The constant 1C peak was raised by round and elongating spermatids (steps 1–14), 2C by spermatogonia, secondary spermatocytes and Sertoli cells, and the 4C peak by primary spermatocytes and spermatogonia at G2 or M phase of the mitotic cycle. The proportion of each peak accurately reflected the relative proportion of cells in most stages of the cycle when compared with morphometric measurements of histologic preparations. DNA flow cytometry is a suitable method for quantitative evaluation of cultured seminiferous tubule segment DNA. Although the relative yield of the meiotic reductive divisions in vitro is comparable with that observed in vivo, steps 9 and 15 of spermiogenesis involving nucleoprotein transitions and spermiation itself did not occur under the present culture conditions.
Differential Release of Polyamines by Cultured Rat Sertoli Cells
Authors: TSAI YU‐HUI , LIN SHEN‐NAN
Published: December 2015

Cellular and media concentrations of polyamines in Sertoli cell cultures were determined by fluorescent spectroscopy of dansylated compounds after separation by high‐performance liquid chromatography. In spite of low cellular levels of putrescine, the Sertoli cells released relatively large amounts of putrescine and spermidine even after several media changes. The inclusion in the culture media of cortisol, insulin, and thyroxine significantly elevated cellular polyamine levels, altered the spermidine to spermine ratio, and enhanced putrescine release by 3‐ to 4‐fold. No spermine, however, was detected in the media under any of the conditions studied. The polyamine concentrations in cultured Sertoli cells from 13‐day‐old rats and the pattern of polyamine release by these cells differed significantly from those in the Sertoli cells from 46‐day‐old rats. These data demonstrate the differential release of polyamines by cultured rat Sertoli cells. The profiles of polyamine secretion appear to be age‐dependent, and the significance of this phenomenon is discussed.
The Effect of Δ9‐tetrahydrocannabinol In Utero Exposure on Rat Offspring Fertility and Ventral Prostate Gland Morphology
Authors: AHLUWALIA BALWANT S. , RAJGURU SHAKUNTALA U. , NOLAN GEORGE H.
Published: December 2015

Male rats exposed in utero to Δ9‐tetrahydrocannabinol (Δ9‐THC) had lower levels of testosterone (T) and luteinizing hormone (LH) prior to puberty (P < 0.01). At puberty, the levels returned to within the normal range. Ultrastructural examination of the ventral prostate gland at puberty revealed alterations suggestive of degenerative changes. A drastic reduction in secretory granules and acini reflected depressed androgen production and function during the developmental period. The fertility of the F1 and F2 male offspring was decreased by 30 to 40%. It is concluded that THC exposure in utero caused a permanent reduction in fertility and altered ventral prostate gland morphology.
Polarized Sertoli Cell Functions in a New Two‐compartment Culture System
Authors: JANECKI ANDRZEJ , STEINBERGER ANNA
Published: December 2015

Sertoli cells in vivo are highly polarized and interact with the inner (tubular) and outer (interstitial) fluids. To simulate this condition in vitro we developed a two‐compartment culture system in which confluent Sertoli cell monolayers were grown on permeable supports (Millipore filters) separating the inner and outer fluid compartments. Monolayer permeability to (3H)‐inulin decreased by 90% after 5 to 7 days of culture, presumably due to formation of tight junctions. This process was influenced by cell plating density. The cells were highly polarized morphologically, resembling their appearance in vivo, and secreted transferrin bidirectionally into both fluid compartments. The amount of transferrin secreted was 166% to 250% of that secreted by the same number of Sertoli cells cultured in plastic dishes. Testosterone (5 × 10−8 M) doubled and testosterone + FSH (0.1 μg/ml) increased transferrin secretion 3.6‐fold. These results demonstrate that under suitable culture conditions the Sertoli cells remain both morphologically and functionally polarized, reflecting a more physiologic state.
Reversibility of Azoospermia Induced by Bacillus Calmette‐Guerin
Authors: NAZ R. K. , TALWAR G. P.
Published: December 2015

A single intratesticular injection of bacillus Calmette‐Guerin induces azoospermia within 3 to 6 weeks in a variety of animals without loss of androgens. After a period of azoospermia, the return of spermatogenesis was observed in dogs and monkeys. Seven dogs that showed aspermatogenesis after a local instillation of bacillus Calmette‐Guerin (20–110 units per testis) again demonstrated spermatogenesis after a period of 153 to 325 days of azoospermia. Sperm counts and motility were restored to preimmunization levels within 6 weeks after initiation of spermatogenesis. In one dog, systemic treatment with antimycobacterial drugs helped to clear the bacillus Calmette‐Guerin from the testis, thus hastening the reinitiation of spermatogenesis. In two monkeys injected with 110 and 160 units of bacillus Calmette‐Guerin, azoospermia lasted up to 150 days. Subsequently, fertility was restored and the treated animals sired healthy offspring. A second injection of bacillus Calmette‐Guerin of a lesser dose (5–20 units per testis) in these fertile dogs and monkeys again produced azoospermia. Histologically, the monkeys having a second period of azoospermia demonstrated aspermatogenic orchitis in the testis and leukocyte infiltration in the interstitium. A majority of the tubules had intact basement membranes, with a block of spermatogenesis at the spermatocyte stage. In dogs, the second azoospermia lasted for 113 to 277 days with a subsequent restoration of sperm counts and sperm motility to preimmunization levels. There were no circulating antisperm antibodies detected in sera of these animals during the azoospermic period or during its reversal. These observations suggest that the bacillus Calmette‐Guerin‐induced azoospermia is reversible, both in terms of sperm count and fertility.
Testicular and Serum Testosterone Variations in Squirrel Monkeys During Seasonal Cyclicity
Authors: PASQUALINI TITANIA , COLILLAS ORESTE , RIVAROLA MARCO A.
Published: December 2015

The seasonal testicular maturation of squirrel monkeys (Saimiri sciureus) was used as a model of maturational hormonal regulation of the testis. Testicular testosterone and serum testosterone concentrations were determined during the circannual variations of body weight and testicular volume. These data have been correlated with changes in the germinal epithelium.

According to individual weight curves and time of the year, the monkeys were divided into five groups: group A1, maximal weight, April‐May; A2, July; A3, November; A4, minimal weight, February‐March; and A5, March‐April. Variations in testicular volume followed very closely variations in body weight. Sexual activity started at A1 and persisted in A2. A marked drop in the mean width of the germinal epithelium and the diameters of the seminiferous tubules was observed in A3, followed by a recovery during A4 and A5.

Testicular testosterone showed two annual elevations. The first peak, 3.91 ± 0.31 μg/g (mean ± SE), coincided with the serum testosterone peak when body weight and testicular volume were high and the trophic response of the germinal epithelium was complete. The second peak reached levels of 5.21 ± 1.48 μg/g and was observed before the reinitiation of spermatogenesis. This was accompanied by a moderate increase in serum testosterone.

The second peak of testicular testosterone, which has been reported to occur in the rat and in humans, might represent a local androgen need for initiation of spermatogenesis, while the first peak might represent the androgen need for full stimulation of spermatogenesis.
Age‐related Variation in Seminiferous Tubules in Men A Stereologic Evaluation
Authors: JOHNSON LARRY , PETTY CHARLES S. , NEAVES WILLIAM B.
Published: December 2015

Tubular boundary tissue and seminiferous epithelia were evaluated stereologically in testes from 28 men aged 20 to 48 years and 28 men aged 50 to 90 years. Testes obtained at autopsy within 15 hours of death were perfused with glutaraldehyde, embedded in Epon (Ladd Research Industries, Inc., Burlington, VT), sectioned at 0.5 μm, and stained with toluidine blue. Volume densities (percentage of the testicular parenchyma) of various parameters determined by point counting and diameter measurements were used to calculate total volumes, length of tubules, and number of cells. Electron microscopy was used to determine the volume density of myoid cells in the boundary tissue. Significant (P < 0.01) age‐related reductions occurred in paired testicular weights, paired parenchymal weights, total volume of seminiferous tubules and of seminiferous epithelium, and length of tubules. The volume density and thickness of boundary tissue increased (P < 0.01) with age. The volume of boundary tissue per man and the volume density of myoid cells in the boundary tissue did not vary with age. Although the number of myoid cells per man tended to be lower in the older group, the number of myoid cells per cross section of seminiferous tubule was increased (P < 0.01) in older men. The age‐related thickening of the boundary tissue was not due to an increase in boundary tissue but resulted from a reduction in the length of the seminiferous tubules.
Seminal Carnitine Content in Obstructive Azoospermia Correlation With the Anatomic Level of Obstruction
Authors: TOMAMICHEL GIAN R. , BANDHAUER KLAUS
Published: December 2015

Free carnitine in human semen originates predominantly in the epididymis. The role of carnitine in the evaluation of different forms of obstructive azoospermia was studied in 42 patients. In 14 of the men, a bilateral vasectomy had been performed. In the remaining 28 patients, the occlusion was located within the epididymis. In postvasectomy cases and where the occlusion was located in the cauda epididymidis, carnitine concentrations were low, with mean values of 115.57 μmol/l and 121.28 μmol/l, respectively. When the occlusion was located in the corpus epididymidis, the mean value increased to 194.72 μmol/l. In patients having obstruction of the caput epididymidis or of the rete testis, the mean value of free carnitine was 416.0 μmol/l. After vasovasostomy, a return of free carnitine concentration to the normal range was observed in 10 of 12 cases. The results indicate that there is a significant correlation in patients with obstructive azoospermia between the concentration of free carnitine and the anatomic site of the obstruction. These findings may lead to important conclusions concerning therapy and prognosis for patients presenting with this condition.
Decrease in the Number of Human Ap and Ad Spermatogonia and in the Ap/Ad Ratio with Advancing Age New Data on the Spermatogonial Stem Cell
Authors: NISTAL MANUEL , CODESAL JAVIER , PANIAGUA RICARDO , SANTAMARIA LUIS
Published: December 2015

The numbers of Ap and Ad spermatogonia per unit section of the testis were calculated in autopsy specimens from young adults and elderly men without testicular pathology. The number of Ap spermatogonia decreased from the 6th decade of life, whereas that of Ad spermatogonia began to decrease in the 8th decade. Although it has been reported that Ad spermatogonia are more sensitive to noxious agents than Ap spermatogonia, the involution of Ap spermatogonia precedes that of Ad spermatogonia. These findings provide new information on concepts relating to the spermatogonial stem cell in man.
Reproductive Responsiveness to Short Photoperiod Develops Postnatally in Male Golden Hamsters
Authors: SISK CHERYL L. , TOREK FRED W.
Published: December 2015

The development of the reproductive response to short photoperiod in golden hamsters was examined by monitoring testis size in hamsters that were either: 1) raised on 14L:10D from birth; 2) raised on 6L:18D from birth; or 3) born on 14L:10D and transferred to 6L:18D at either 1, 2, 3, 4, 5, 6, 7, 9, or 12 weeks of age. Testis growth occurred between 3 and 7 weeks of age at similar rates in all groups. Testicular regression occurred in all groups exposed to short days; the time of regression depended on the age of the animals when they were first transferred to 6L:18D. Testicular regression began at about 10 weeks of age in all groups transferred to 6L:18D between birth and 5 weeks of age. In contrast, regression began at 11.2, 13.3, 15.6, and 18.2 weeks of age in hamsters transferred to 6L:18D at 6, 7, 9, and 12 weeks of age, respectively. These results suggest that the reproductive system of male hamsters is refractory to the inhibitory effects of short days on reproductive function until 5 to 6 weeks of age. Thereafter, exposure to short days initiates testicular regression in hamsters within 5 to 7 weeks.
Reversibility of Long‐term Effects of GnRH Agonist Administration on Testicular Histology and Sperm Production in the Nonhuman Primate
Authors: WEINBAUER GERHARD F. , RESPONDEK MICHAEL , THEMANN HERMANN , NIESCHLAG EBERHARD
Published: December 2015

The present investigation evaluates the long‐term effects of GnRH agonist treatment on testicular histology, sperm production and the subsequent recovery of these parameters. Four adult rhesus monkeys (M. mulatta) were treated with the GnRH agonist nafarelin (D‐Nal(2)6‐GnRH), released from i.m.—injected poly‐D,L‐lactic‐coglycolide microspheres for 20 months. Monthly injection of the GnRH agonist preparation uniformly suppressed serum levels of bioactive LH and testosterone. The size of the testis was reduced to about 30% of pretreatment. Sperm counts were suppressed to azoospermia for a total period of 53 and 77 weeks, respectively, in two monkeys and the other two animals were extremely oligozoospermic. Evaluation of testicular biopsy material after 6, 12 and 20 months of treatment revealed decreased seminiferous tubule diameter, spermatogenic disruption at the level of spermatogonia or spermatocytes, accumulation of lipid droplets and secondary lysosomes in the Sertoli cell cytoplasm, and increased thickness of the tubular wall compared with pretreatment histology. Electron microscopic examination revealed that the increased wall thickness was due to an enlargement of the inner collagen layer. No evidence of fibrosis or calcification could be obtained. Leydig cells were atrophic. Serum hormones, testis size and sperm counts returned to pretreatment values within 5 to 8, 13 to 16, and 18 weeks, respectively, after termination of treatment. Testicular histology, assessed 8 months after cessation of treatment, was indistinguishable from pretreatment. It is concluded that GnRH agonist‐containing microspheres are a feasible modality for sustained administration of GnRH agonists and GnRH agonist‐induced suppression of pituitary and testicular function is reversible following withdrawal of treatment. Thus, GnRH agonists may have a potential for regulation of male fertility and, presumably, also for treatment of precocious puberty.
Pituitary‐Testicular Function of Prostatic Cancer Patients During Treatment with a Gonadotropin‐Releasing Hormone Agonist Analog II. Endocrinology and Histology of the Testis
Authors: HUHTANIEMI Dr.I. , NIKULA H. , PARVINEN M. , RANNIKKO S.
Published: December 2015

Endogenous testosterone (T), LH and FSH receptors, and in vitro production of cyclic adenosine‐3′:5′‐monophosphate (cAMP), T and some of its steroid precursors were measured in testicular tissue obtained at orchiectomy from seven prostatic cancer patients treated for 6 months with a potent gonadotropin‐releasing hormone (GnRH) agonist analog (buserelin, Hoechst, 600 μg 3 times a day intranasally). In addition, histologic and morphometric studies were carried out on the testicular tissue. Testicular tissue from age‐matched prostatic cancer patients (n = 14), whose first therapy was orchiectomy, served as controls. The peptide treatment decreased intratesticular T by 95% (P < 0.01) and FSH receptors by 57% (P < 0.01), but had no effect on LH receptors. The in vitro production of T decreased by 94% (P < 0.01), but that of cAMP was unaffected. Besides T, the in vitro production of testicular 17‐hydroxyprogesterone (17‐OHP‐4), androstenedione and 5α‐dihydrotestosterone (5α‐DHT) dropped by 71 to 90% (P < 0.01 to 0.05) during buserelin treatment, but those of pregnenolone, progesterone and dehydroepian‐drosterone (DHEA) were not affected. Histologic studies revealed considerable variation in the seminiferous epithelium of the control group, but spermatogenesis was highly suppressed in nearly all of the buserelin‐treated group. The number of Sertoli cells was unaffected, but tubular diameters were reduced (P < 0.05) by buserelin treatment. Leydig cells appeared dedifferentiated in this group, although their number per testis was not altered. These data indicate that gonadotropin suppression by GnRH agonist most likely affects testicular steroidogenesis by inhibiting 3β‐hydroxysteroid dehydrogenase and a step(s) prior to pregnenolone formation. The treatment does not impair testicular LH binding or cAMP production, but clearly suppresses FSH receptors. Spermatogenesis in general is suppressed but with considerable variation.
Characteristics of Prolactin‐Modulated LH Induction of LH/hCG Receptors: Transient Inhibition of Receptor Induction Following Prolactin Exposure
Authors: HUSSEIN MOHAMED O. , ZIPF WILLIAM B.
Published: December 2015

The temporal relationship between exposure to prolactin (PRL) and luteinizing hormone (LH) induction of LH receptors was investigated in hypophysectomized adult male rats. Testicular homogenate membrane preparations were incubated with [125I]hCG for analysis of LH/hCG binding. Seven days after hypophysectomy, the rats were injected with 100 μg/day of PRL for another 7 days and then given a single 10‐μg dose of LH at 2, 4, 6, 12, 24, or 36 hours after the last PRL injection. The priming effect of PRL on LH induction of receptors was not observed if LH was administered 2 to 12 hours from the last PRL injection. However, after this inhibitory period, injections of LH to PRL‐primed rats resulted in induction of LH receptors and the effect persisted for 36 hours. This study supports previous reports demonstrating a unique dependence upon PRL for LH up‐regulation of the LH receptor and characterizes the brief refractory period following exposure to PRL.
Male Fertility and Positive Chlamydial Serology A Study of 61 Fertile and 82 Subfertile Men
Authors: AUROUX MAURICE R. , de MOUY DANNY M. , ACAR JACQUES F.
Published: December 2015

In this study, the frequency and titers of serum antibodies to Chlamydia trachomatis in fertile and subfertile subjects and possible correlations between a positive serology and the values of conventional semen parameters were investigated. The subfertile subjects were divided into those without and those with a history of genitourinary infection. No difference was noted between the proportion of seropositive patients in the fertile group (26.2%) as compared with those in the subfertile group with no history of genitourinary infection (27.7%). The percentage of positive tests was significantly increased only in the subfertile group with past genitourinary infection (53.5%). No difference was found between the antibody titers of the fertile and subfertile groups. Only the fertile group had any semen parameter, namely, the volume of the ejaculate, that was significantly different between the seropositive and seronegative patients. In conclusion, three main points should be stressed: 1) the high frequency of serum antibodies to Chlamydia trachomatis in the control subjects and the apparent absence of effects of a positive serology on their fertility; 2) the significant correlation between a past genitourinary infection and the presence of serum antibodies to Chlamydia trachomatis; and 3) the need for a test to assess Chlamydia in semen.
Temporal Appearance of Antisperm Autoantibodies in Lewis Rats Following Vasectomy
Authors: HERR JOHN C. , FLICKINGER CHARLES J. , HOWARDS STUART S. , YARBRO E. SCOTT , SPELL DONALD R. , CALORAS DANIEL , GALLIEN THOMAS N.
Published: December 2015

An indirect enzyme‐linked immunosorbent assay (ELISA) was employed to monitor antisperm autoantibodies in 16 Lewis rats for up to 36 weeks following vasectomy. This assay was capable of discriminating all prevasectomy from postvasectomy sera at a 1:16 dilution. Weekly serum samples were obtained for the first 13 weeks and bimonthly samples thereafter. Half of the animals developed a positive antisperm autoantibody response by the end of the first postoperative week. By the end of the second week, 81% of the animals had positive responses. The greatest proportion (88%) of animals having a positive response over the course of the study was found at the end of the seventh postoperative week and the highest mean absorbance value for all 16 animals was observed at this time. Only 25% of the animals had positive responses for antisperm autoantibody at the end of the 35th week of the study. These findings indicate that circulating antisperm autoantibodies arise in the Lewis rat earlier than has been generally appreciated. The time course is similar to that of antibody titers to infectious agents or arising from inoculation of rats with spermatozoa. These findings on autoantibody levels in the Lewis rat are compared with the dynamics of antisperm autoantibody formation in man.