bluuebunny/crossref_metadata_2025 huggingface dataset:
Mechanism of Action of Gonadotropin‐releasing Hormone‐stimulated Leydig Cell Steroidogenesis.
Authors: LIN TU
Published: December 2015

To investigate mechanisms responsible for gonadotropin‐releasing hormone (GnRH)‐stimulated Leydig cell steroidogenesis, the effects of GnRH agonist [des‐Gly10, (D‐Ala6) GnRH] on phospholipid turnover were studied. GnRH agonist in concentrations of 10−9 to 10−7 M increased phosphatidic acid labeling 292 ± 16% (mean ± SE), and phosphatidylinositol labeling 258 ± 13.2%. GnRH agonist‐stimulated phospholipid labeling was detectable as early as 2 minutes. GnRH antagonist completely blocked GnRH agonist‐induced testosterone formation and phosphatidic acid and phosphatidylinosital labeling. Nifedipine in concentrations of 1 and 10 μg/ml inhibited GnRH agonist‐stimulated testosterone formation but had no effect on 32P incorporation into phospholipids. Our results suggest that GnRH agonist‐stimulated Leydig cell steroidogenesis is calcium dependent and correlated with increased phospholipid turnover.
Intranasal Luteinizing Hormone Releasing Hormone (LHRH) Therapy in Cryptorchid Children
Authors: BERNASCONI SERGIO , VANELLI MAURIZIO , ROSSI STEFANO , GHINELLI CESARE , ZIVERI MARIANGELA , VIRDIS RAFFAELE
Published: December 2015

The clinical and hormonal response to synthetic intranasal LHRH therapy (3 × 400 μg daily for 28 days) was evaluated in 23 unilateral and nine bilateral cryptorchid children. Complete descent was obtained in 14 cryptorchid testes (34%), whereas in seven testis there was only an improvement in the original position. GnRH test, performed before and after therapy, did not show any significant difference between responder and nonresponder subjects. FSH peak levels, however, decreased in all groups of patients after treatment. LH and FSH levels were monitored before, during (days 7 and 14), and after (day 28) therapy. FSH and LH values increased during therapy both in responders and nonre‐sponders, returning to baseline at the end of the treatment. Testosterone remained constant throughout the period of investigation. The results obtained and the absence of side effects indicate that this therapeutic approach may be seen as the treatment of choice for cryptorchidism.
Varicocele and Infertility
Authors: FORESTA CARLO , RUZZA GRAZIA , RIZZOTTI ALBERTO , LEMBO ANTONIO , VALENTE MARIA LUISA , MASTROGIACOMO ISMAELE
Published: December 2015

In order to develop a procedure permitting a more accurate preoperative prognosis of varicocelectomy, we studied 59 subjects (20 to 40 years of age) who had undergone ligation of the spermatic vein for varicocele and infertility at least two years previously. The following preoperative parameters were evaluated: age, seminal fluid, plasma levels of LH and FSH and their maximal increase after LHRH stimulation, plasma levels of testosteone and its maximal increase after hCG (5000 U) administration, and testicular histology. Patients who failed to impregnate their wives after surgery were older, had higher plasma levels of FSH and a higher maximal increase in FSH and LH after LHRH stimulation, as well as more compromised testicular histology. Plasma levels of testosterone and seminal characteristics did not appear to constitute valid prognostic factors.
Induced Hypoprolactinemia and Testicular Steroidogenesis in Man
Authors: SUESCUN MARÍA O. , SCORTICATI CARLOS , CHIAUZZI VIOLETA A. , CHEMES HÉCTOR E. , RIVAROLA MARCO A. , CALANDRA RICARDO S.
Published: December 2015

The effects of short‐term hypoprolactinemia on the pituitary‐gonadal axis were evaluated in a group of patients with untreated prostatic carcinoma. Each patient was studied prior to and during 7‐day oral administrations of bromocriptine (2.5 mg q.i.d.). Serum LH, prolactin (PRL), androst‐4‐ene‐3,17 dione (androstenedione), testosterone, and 5α‐androstane‐3α,17β‐diol (5α‐Diol) levels, as well as intra‐testicular testosterone, dihydrotestosterone (DHT), 5α‐Diol and zinc (Zn) concentrations, were determined. Daily administration of bromocriptine caused a marked supression of serum PRL (mean ± SEM, 23.8 ± 2.5 vs. 6.4 ± 1.0 ng/ml) without concomitant changes in serum LH levels (mean ± SEM, 8.3 ± 1.6 vs. 8.9 ± 2.1 ng/ml). Hypoprolactinemia induced a significant decrease (P < 0.05) in the mean peripheral testosterone levels; but 5α‐Diol and androstenedione remained unchanged. However, in testicular tissues, bromocriptine treatment resulted in significant increases in mean concentrations of total androgens (P < 0.001), testosterone (P < 0.001) and DHT (P < 0.02). Testicular levels of 5α‐Diol were not significantly altered. There was no change in Zn levels in basal conditions and during bromocriptine administration. These results indicate that short‐term suppression of serum PRL levels in man affects basal testicular function without altering serum LH. However, a direct action of bromocriptine on the human gonad cannot be excluded.
Prolonged Suppression of Plasma LH Levels in Male Rats after a Single Injection of an LH‐RH Agonist in Poly(DL‐Lactide‐Co‐Glycolide) Microcapsules
Authors: ASCH RICARDO H. , ROJAS FRANCISCO J. , BARTKE ANDRZEJ , SCHALLY ANDREW V. , TICE THOMAS R. , KLEMCKE HAROLD G. , SILER‐KHODR THERESA M. , BRAY ROWENA E. , HOGAN MARY P.
Published: December 2015

The authors have examined the effects of a subcutaneous injection of the LH‐RH agonist D‐Trp6‐LH‐RH formulated in biodegradable poly(DL‐lactide‐co‐glycolide) microcapsules on plasma levels of D‐Trp6LH‐RH, LH, and PRL in adult, gonadectomized male rats. Immunoreactive D‐Trp6‐LH‐RH was detectable in the plasma of these animals at 1, 2, 3, and 4 weeks after injection. LH concentrations were greatly reduced 1 week after administering the D‐Trp6‐LH‐RH microcapsule, continued to decrease during the following week, and remained suppressed until the end of the study, 6 weeks after the injection. Plasma PRL levels appeared elevated 1 to 2 weeks after the injection and suppressed thereafter, but these effects were significant only in animals rendered hyperprolactinemic by transplantation of an isologous pituitary under the renal capsule. These results demonstrate that an LH‐RH agonist formulated in biodegradable microcapsules and administered as a subcutaneous injection can exert marked biologic effects in rats for at least 6 weeks. These findings also suggest that prolonged exposure to an LH‐RH agonist may first produce stimulation, followed by an inhibition of PRL release from both in situ and ectopic pituitaries.
Prostaglandin Dehydrogenase Activity of Rat and Rabbit Testicular Tissues and Accessory Glands before and after Castration
Authors: NEMETALLAH BAHIG R. , ELLIS LEGRANDE C.
Published: December 2015

Rat reproductive tissues contained both 15‐hydroxyprostaglandin dehydrogenase (PGDH) and Δ13‐reductase activities, while rabbit tissues exhibited only PGDH activity. In the rat, Δ13‐reductase activity was absent in those tissues that had a high specific activity of PGDH and was present in those tissues with a low specific activity. Total specific activity of PGDH was greatest in the testicular capsule, whereas total activity was greatest in the testicular parenchyma of both species. Total PGDH activity was highest in rat seminal vesicles, where it was second only to the testicular parenchyma. Castration significantly increased PGDH activity of the epididymis, prostate, and seminal vesicles of rats, while Δ13‐reductase activity disappeared from those tissues. The specific activity of PGDH was greater in the rat than in the rabbit, except for the testicular parenchyma of the rabbit. The greater PGDH activity in rat testicular capsules, compared to those of the rabbit, correlated well with diminished contractility of the rat capsule. PGDH activity of the interstitial cells correlated well with the role of prostaglandins in androgen synthesis.
Stagnation of Blood in the Microvasculature of the Affected and Contralateral Testes of Men with Short‐term Torsion of the Spermatic Cord
Authors: CHAKRABORTY JYOTSNA , HIKIM AMIYA P. SINHA , JHUNJHUNWALA JAGADISH S.
Published: December 2015

Bilateral testicular biopsies from four men with a short duration (3 hours 10 minutes to 4 hours 30 minutes) of unilateral spermatic cord torsion and testicular biopsies from six men with irreversible brain death were used for the present investigation. Extensive light and electron microscopic studies and quantitative analyses of all biopsy materials were performed. The torsioned testes revealed variable degrees of damage to the seminiferous tubules, including germ cell disorganization and sloughing of immature germ cells. Ninety‐five percent of the blood vessels from the biopsied tissue specimens were clogged with blood cells. The seminiferous tubules of the contralateral testis had normal germ cell arrangements and counts. However, 88% of the microvessels from the tissue biopsied from the contralateral testes were packed with blood cells, whereas only 10% of the blood vessels in the control biopsy specimen were clogged with blood cells. At the electron microscopic level, fewer tight junctions and enlarged pores were found between the endothelial cells of the affected vessels, and microvilli were completely absent from these endothelial cells. The clogging caused by blood cells in the affected vessels was so severe that no space was found between the membrane of the endothelial cell and the membrane of the blood cells. It has been suggested that local clogging by blood is responsible for the initiation of degenerative changes in the testes of men with unilateral torsion of the spermatic cord.
Growth and Characterization of Polarized Monolayers of Epididymal Epithelial Cells and Sertoli Cells in Dual Environment Culture Chambers
Authors: BYERS STEPHEN W. , HADLEY MARK A. , DJAKIEW DANIEL , DYM MARTIN
Published: December 2015

Epididymal epithelial cells isolated from mature rats and Sertoli cells isolated from 10‐day‐old rats were cultured in serum‐free defined media on extracellular matrix impregnated filters maintained in dual environment culture chambers. Epididymal epithelial cells had a polarized appearance only when plated at high density (> 1 × 106 cells/cm2). Confluent monolayers of these cells formed a permeability barrier to inulin. Sertoli cells were columnar and highly polarized when grown on extracellular matrix‐impregnated filters, cuboidal when grown on filters alone, and squamous when grown on plastic. Confluent polarized monolayers of these cells excluded the electron‐dense tracer lanthanum nitrate by way of basal‐tight junctions. Therefore, polarized monolayers of epididymal epithelial cells and Sertoli cells can be obtained by growing the cells at high density on extracellular matrix‐impregnated permeable supports. By maintaining the monolayers in specially constructed culture chambers, the cells can develop a permeability barrier, and are able to achieve the separation of apical from basal compartments so important for their function in vivo.
American Society of Andrology Its Beginnings
Authors: ROSEMBERG EUGENIA
Published: December 2015

The term “andrology” was introduced in 1951 by Harald Siebke, a Professor of Gynecology in Bonn, Germany. However, many years passed before the term gained acceptance. It was not until 1969 when, due to the efforts of Dr. Carl Schirren of Hamburg, Germany that the first scientific journal dedicated to the subject titled Andrologie initiated its publication in West Germany (Schirren, 1985). Andrology now is recognized as an area of science and medicine which fosters a multidisciplinary and multifaceted approach to the study of male reproduction. Andrology encompasses both basic and clinical sciences. It includes research in biochemistry, genetics, histology, immunology, molecular biology, pathology, pharmacology, physiology, and endocrinology. It also includes urology, microsurgery, gynecology, internal medicine, pediatrics, psychology, and animal husbandry.

The American Society of Andrology came into being not as an isolated development but as a consequence of events promoted by individuals deeply committed to the study of male reproduction. The first of these events took place in the U.S.A. when Drs. W. O. Nelson and C. H. LeBlond suggested that a club of scientists interested in the study of male reproduction be organized. At that time, Dr. E. Steinberger became actively involved in the organization of the club, which held its first meeting in 1968. The club became known as the Warren O. Nelson Club, and held meetings for the following four years. Although it was proposed that the club become a society, the idea did not materialize.
Identification and Isolation of Epididymal Luminal Proteins of the Mouse
Authors: FLICKINGER CHARLES J. , HERR JOHN C. , ERTL KARIN E.
Published: December 2015

Proteins of mouse cauda epididymal fluid were analyzed by polyacrylamide gel electrophoresis. Fluid expressed from the cauda epididymidis and samples obtained by micropuncture of the epididymal lumen showed very similar patterns with respect to the major proteins they contained, with the exception of a small amount of serum albumin found in expressed caudal fluid. Eight prominent peptides present in both micropuncture fluid and expressed caudal epididymal fluid were selected for further study, and were designated CP 47, 42, 35, 29, 27, 25,18, and 13 according to their mobility. Six of these were never detected in serum. Periodic acid‐Schiff staining indicated that at least three were glycoproteins. The epididymal proteins were purified by preparative polyacrylamide gel electrophoresis and electroelution. Upon re‐electrophoresis, the individual purified peptides comigrated with the corresponding bands in whole epididymal fluid, and no additional bands were detected, indicating that the proteins were purified to a high degree of homogeneity. Several of the mouse epididymal peptides resemble in their mobility proteins identified previously in other species, most notably the widely studied 33 Kd and 16 to 18 Kd proteins detected in the rat.
Kinetic Characterization of the Inhibition of Purified Cynomolgus Monkey Lactate Dehydrogenase Isozymes by Gossypol
Authors: STEPHENS DONALD T. , WHALEY KEVIN J. , KLIMKOW NANETTE M. , GOH PAULINE , HOSKINS DALE D.
Published: December 2015

This report describes the results of the first step in a sequence of experiments designed to test the hypothesis that the sperm‐specific isozyme of lactate dehydrogenase (LDH‐C4), is a site of action of the potential male contraceptive agent gossypol. Cynomolgus monkey LDH‐A4, LDH‐B4, and LDH‐C4 were purified and kinetically characterized. LDH‐A4 and LDH‐B4 exhibited “linear mixed‐type” inhibition by gossypol with both lactate and pyruvate as variable substrates. LDH‐C4 also exhibited “linear mixed‐type” inhibition with lactate as substrate. However, the C4 isozyme exhibited “parabolic mixed‐type” inhibition by gossypol and substrate inhibition with pyruvate as substrate, the latter due to abortive complex formation. Of the three isozymes, LDH‐C4 exhibited the lowest apparent Km for pyruvate and the highest apparent Km for lactate. The LDH‐C4 form was found to be the most sensitive isozyme to gossypol inhibition, since it had the lowest apparent K1 values for gossypol inhibition. The effect of gossypol on coenzyme binding to LDH‐C4 was examined and gossypol binding was found to inhibit binding and release of NADH but not NAD+, an effect possibly due to its interaction with the more hydrophobic loop region of LDH‐C4.
Follicle Stimulating Hormone, Luteinizing Hormone and Testicular Leydig Cell Responses to Estradiol Immunization in Ile‐de‐France Rams
Authors: SCHANBACHER B. D. , PELLETIER J. , REVIERS M. T. HOCHEREAU‐DE
Published: December 2015

Active immunization of Ile‐de‐France rams against estradiol (E2) resulted in the production of E2‐neutralizing antibodies and an elevation in the plasma concentrations of FSH, LH, and testosterone. The presence of E2 antibodies did not affect the testosterone metabolic clearance rate, indicating that the immunization‐mediated 10‐fold increase in plasma testosterone was the result of a 10‐fold increase in testicular testosterone production. Testis weights, as well as nuclear and cytoplasmic volumes of individual peritubular and perivascular Leydig cells, were greater in E2‐immunized rams than in albumin‐immunized controls. Leydig cell numbers were not affected by treatment. The E2 antibodies were capable not only of neutralizing the inhibitory effects of endogenous E2 on gonadotropin levels in intact rams, but were able to block the effects of exogenously administered E2 on their FSH and LH secretory response to castration. It is concluded that circulating E2 in the ram is involved in pituitary‐testicular endocrine homeostasis and that E2 immunoneutralization can be employed to enhance testosterone secretion in this species.
The Effects of Unilateral, Experimental Varicocele Are Not Mediated Through the Ipsilateral Testis
Authors: HURT G. S. , HOWARDS S. S. , TURNER T. T.
Published: December 2015

Unilateral varicocele has been associated with diminished male fertility in humans and with bilateral physiologic and histologic changes in the testis of humans and laboratory animals. In particular, left varicocele in Sprague‐Dawley rats results in bilateral increases in testicular temperature and blood flow. The mechanism by which unilateral varicocele can cause testicular changes is not known. The purpose of the present study was to determine whether or not the presence of either the ipsilateral or contralateral testicle is necessary for these effects of the varicocele to occur in the opposite testis. Varicoceles were created in adult, male rats by partial constriction of the left renal vein. Bilateral testicular blood flow was measured by a radiolabelled microsphere distribution technique and testicular temperature was taken with a needle probe thermometer. Right or left orchiectomies were performed on selected animals at the time of surgery to establish the unilateral left varicocele. Animals were studied 30 days after surgery. Mean testicular blood flow was significantly increased (P < 0.01) in all animals having a left varicocele when compared with animals not having a varicocele regardless of whether a unilateral orchiectomy was performed. Likewise, the mean difference between intraabdominal temperature and intratesticular temperature (Δ T) was significantly decreased in all groups of animals having varicoceles when compared with groups without varicoceles whether or not an orchiectomy had been performed. Thus, the studied bilateral effects of left‐sided, experimental varicocele in the rat are not dependent upon the presence of a left testicle.
Appraising the Instantaneous Secretory Rates of Luteinizing Hormone and Testosterone in Response to Selective Mu Opiate Receptor Blockade in Late Pubertal Boys
Authors: MAURAS NELLY , ROGOL ALAN D. , VELDHUIS JOHANNES D.
Published: December 2015

The pulsatile properties of gonadotropin and testosterone release were examined before and after chronic mu opiate receptor blockade with naltrexone, 50 mg every other day, in four normal boys in late puberty (ages 14 8/12 to 15 1/12 years). The nature of spontaneous secretory events was appraised for immunoactive LH and testosterone in blood withdrawn every 20 minutes for 24 hours, using a novel, discrete deconvolution algorithm to estimate apparent instantaneous secretory rates. The application of this methodology revealed that the frequency of discrete LH instantaneous secretory rates increased after mu opiate receptor blockade (P = 0.011). More strikingly, all parameters of testosterone secretory events responded significantly to mu opiate receptor blockade, including increases in mean estimated secretory rate (+ 47%, P = 0.02), testosterone pulse frequency (+ 64%, P < 0.001) and amplitude (+ 20%, P = 0.027). Correspondingly, decreases in testosterone interpulse secretory intervals (‐ 35%, P = 0.001), secretory pulse duration (‐ 19%, P = 0.042) and interpulse valley duration (‐ 35%, P = 0.006) also were noted. There was a prominent diurnal rhythm in testosterone secretion with maximal values in the morning and late evening, and marked reductions in the afternoon, sometimes to prepubertal levels. This variation in the testosterone secretory profile paralleled that of LH. In response to naltrexone, the FSH concentration series showed a significant increase in the mean FSH concentration (+ 18%) P = 0.003) and mean peak amplitude (+ 15%, P = 0.002). These data provide indirect evidence of functional coupling of the opiate system with the hypothalamic GnRH pulse generator. The marked increase in LH and testosterone secretory activity (as measured by the instantaneous secretory rate) after opiate receptor blockade suggests an integral interaction between the endogenous opiate system and the hypothalamic‐pituitary gonadal axis during the later stages of puberty in males.
Chronic Sinopulmonary Disease in Chinese Patients with Obstructive Azoospermia
Authors: WANG CHRISTINA , SO S.Y. , WONG K.K. , SO WILLIAM W.K. , CHAN STEVEN Y.W.
Published: December 2015

The prevalence of chronic infections of the nasal sinuses and the lung (chronic sinopulmonary disease) was investigated in 33 Chinese patients with obstructive azoospermia who were compared with 32 patients with azoospermia due to failure of the germinal epithelium. Ten out of the group of 33 patients with obstructive azoospermia had congenital absence of the vas and/or epididymis and were excluded from that group. The patients with obstructive azoospermia had normal testis size, normal levels of serum FSH and LH, lower seminal fluid fructose, and a higher incidence of serum sperm agglutinating and immobilizing antibodies when compared with the group with damage to the germinal epithelium. The number of patients with symptoms of chronic sinopulmonary infections were similar in all groups. One patient with obstructive azoospermia had bronchiectasis. All other patients had normal chest x‐ray studies. About 40% of the patients in all three groups had abnormal sinus X‐rays. However, the nonsmoking patients with obstructive azoospermia had a statistically significant (P <0.05) lower mid‐expiratory flow rate than the nonsmoking patients with nonobstructive azoospermia. Only three additional patients with obstructive azoospermia had both abnormal sinus x‐ray and a reduced mid‐expiratory flow rate, suggestive of Young's syndrome. It was concluded that Young's syndrome (sinopulmonary infections associated with acquired obstructive azoospermia) is much less common in Chinese men (four out of 23, 17%) and their sinopulmonary problems are less severe than in Caucasians.
Alterations in the Pulsatile Properties of Gonadotropin Secretion in Alcoholic Men
Authors: IRANMANESH ALI , VELDHUIS JOHANNES D. , SAMOJLIK EUGENE , ROGOL ALAN D. , JOHNSON MICHAEL L. , LIZARRALDE GERMAN
Published: December 2015

The functional characteristics of the hypothalamic‐pituitary‐testicular axis were examined quantitatively in 10 chronic alcoholic men without hepatic dysfunction or clinical nutritional deficiencies. Spontaneous gonadotropin pulsatility was analyzed in blood sampled every 20 minutes over a 24‐hour period 3 to 16 days after abstinence from alcohol and again 29 to 39 days later. The numbers of LH and FSH pulses per 24 hours were normal in these alcoholic men compared with controls. However, we found increased mean 24‐hour concentrations of immunoactive LH (P = 0.012) and FSH (P = 0.018), increased peak heights for LH (P = 0.035) and FSH (P = 0.004), decreased fractional LH (P = 0.002) and FSH (P = 0.044) pulse amplitudes and increased interpulse valley mean LH (P = 0.010) and FSH (P = 0.018) concentrations. Serum levels of total testosterone, total estradiol and estrone were normal, whereas concentrations of free testosterone and free estradiol were increased. Pituitary release of LH and FSH was normal in response to low (5‐μg) and high (95‐μg) doses of GnRH given intravenously. The present observations indicate that in chronically alcoholic men, acute abstinence from ethanol is associated with elevated circulating concentrations of immunoactive gonadotropins in the presence of intact spontaneous gonadotropin pulsatility, preserved pituitary responsiveness to exogenous GnRH, and increased concentrations of free testosterone and free estradiol. Such findings are consistent with alterations in the endogenous feedback actions of sex steroid hormones in this setting.
Ethanol Accelerates Acrosomal Loss in Human Spermatozoa
Authors: ALVAREZ JUAN G. , LEE MICHAEL A. , IOZZO RENATO V. , LOPEZ ISABEL , TOUCHSTONE JOSEPH C. , STOREY BAYARD T.
Published: December 2015

The effects of ethanol on the loss of the human sperm acrosome, as determined by the chlortetracycline fluorescence assay and by indirect immunofluorescence assay, were assessed over 6 hours during incubation at 37 C in BWW medium containing 0 to 250 mM ethanol. Both assays gave the same results. At the end of 6 hours, 48 ± 6% acrosomal loss was found in samples in 250 mM ethanol compared with 4 ± 1% in the absence of ethanol. After 0.25 hour, the first time point chosen for sampling, the spermatozoa in 250 mM ethanol showed 23 ± 3% loss of acrosomes compared with < 1% in the absence of ethanol. Ultrastructural studies revealed that the ethanol‐treated spermatozoa showed complete acrosomal loss as well as loss of the equatorial segment. No examples of the vesiculation characteristic of the physiologic acrosome reaction were found in the 150 cells examined. Calcium is required for the ethanol‐mediated acrosomal loss: omission of Ca2+, addition of 2 mM EGTA, or 0.2 mM verapamil blocked the effect. Ethanol induced a dose‐dependent efflux of cholesterol from human spermatozoa, but the ethanol‐induced acceleration of acrosomal loss occurred to the same extent in the presence of cholesterol microdispersions that prevented this efflux. The loss of the equatorial segment, which is necessary for egg penetration, during ethanol‐induced acrosomal loss would explain the known effect of ethanol in inhibiting, rather than enhancing, the penetration of zona‐free hamster eggs by human spermatozoa.
Purification and Characterization of a Sperm Motility Inhibitor in Human Seminal Plasma
Authors: IWAMOTO TERUAKI , GAGNON CLAUDE
Published: December 2015

A sperm motility inhibitor from human seminal plasma was purified and characterized. The purification procedure includes dialysis, ion exchange chromatography on SP‐Sephadex C‐25 and adsorption chromatography on hydroxylapatite. With this procedure, the seminal plasma motility inhibitor was purified 290‐fold with a 24% recovery in inhibitory activity. Its molecular weight has been estimated at 18,000 to 22,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, but at 13,000 to 15,000 according to molecular sieving under native conditions. The mobility inhibitor has an isoelectric point pH 9.1. It is stable over a wide range of pH (5 to 10) and at temperatures up to 60 C. The observation that the seminal plasma factor inhibited purified bull dynein ATPase in a concentration‐dependent manner may suggest that it blocks the motility of demembranated spermatozoa by interfering with dynein arm function.
Steroid Metabolism and Morphologic Features of the Human Testis
Authors: FREDRICSSON BENGT , CARLSTRÖM KJELL , PLÖEN LEIF
Published: December 2015

Serum FSH, LH, and testosterone were studied in 114 infertile men with poor sperm production. Testicular biopsies were taken and classified morphologically. In 90 specimens, the pattern of conversion of progesterone was determined in vitro and expressed as the ratio of 20α‐hydroxy‐4‐pregnene‐3‐one to 17α‐hydroxyprogesterone. An excess of the former metabolite indicates a prepubertal type of steroid metabolism. The normal limit of this ratio is given. The collected data indicate that the prepubertal type of steroid metabolic pattern is related to the thickness of the lamina propria of the seminiferous tubules only, and not to peripheral hormone levels. In particular, the presence of hyaline deposits in the lamina propria seems to determine the metabolic pattern. It is suggested that the character of the lamina propria separating the tubular and interstitial compartments of the testis is of crucial importance for the functional interrelationship between these compartments. This supports the concept of an intratesticular regulatory mechanism of both steroid metabolism and spermatogenesis.
Microrecanalization After Vasectomy in Man
Authors: FREUND MATTHEW J. , WEIDMANN JAMES E. , GOLDSTEIN MARC , MARMAR JOEL , SANTULLI ROSEMARY , OLIVEIRA NANCY
Published: December 2015

Previously spermatozoa in the semen of vasectomized men were reported in 62 of 63 specimens from 24 men 2 to 31 years postvasectomy (Freund and Couture, 1982). A morphologic basis and term, “microrecanalization,” was proposed for this observation. Serial sections (5 μ at 200‐μ intervals) of 40 specimens removed at vasovasostomy from 20 men (2 to 14 years postvasectomy) were examined and microcanals (small epithelial‐lined channels) were demonstrated in 27 specimens from 18 men. In nine of the 27 specimens, spermatozoa or sperm heads were found within the microcanals. Microcanals occurred in smooth muscle, connective tissue and scar tissue, in each segment, testicular, central and abdominal, in the presence or absence of the vas deferens. Microcanal continuity was traced for 200 to 1140 microns by computerized image analysis. Microrecanalization is characterized by the absence of inflammation or sperm extravasation and is histologically distinct from vasitis nodes or sperm granuloma. Microrecanalization provides morphologic and physiologic bases for the protection of the testis and maintenance of spermatogenesis in man after vasectomy.
Polyamine Profiles in Rat Testis, Germ Cells and Sertoli Cells During Testicular Maturation
Authors: SHUBHADA SANKARARAMAN , LIN SHEN‐NAN , QIAN ZHI‐YU , STEINBERGER ANNA , TSAT YU‐HUI
Published: December 2015

Polyamine cellular concentrations (putrescine, spermidine and spermine) in the rat testis and testicular cell types were determined by fluorescence spectroscopy of their dansyl derivatives. A method is described to separate dansylated polyamines by high performance liquid chromatography in less than 12 minutes. In rat Sertoli cells, polyamine concentrations (per mg DNA) were greater than those in germ cells and the testis. The concentrations of all three polyamines increased with age. Concentrations of spermidine and spermine in germ cells also increased with age and leveled off after 27 to 35 days. On the other hand, higher putrescine levels were found in the testis of young rats (13 to 22 days) while the greatest spermidine and spermine contents were observed in the testis from rats of 31 to 35 days old. Of great interest, Sertoli cells from all age groups studied released a relatively large quantity of putrescine and a smaller amount of spermidine, but no spermine, into culture media. The amount of polyamine released by Sertoli cells varied with the age of the animal. Sertoli cells from 27‐day‐old rats released the greatest quantity of putrescine on a per mg DNA basis. The release of putrescine increased after hypotonic treatment that removed contaminating germ cells from the remaining Sertoli cells. It is concluded that cellular polyamine levels in the rat testis, germ cells and cultured Sertoli cells and the amount of polyamines released by Sertoli cells were age‐dependent during the first wave of spermatogenesis.
Phosphatidylcholine Synthesis in Human Spermatozoa
Authors: JONES ROBERT E. , PLYMATE STEPHEN R.
Published: December 2015

To clarify the mechanism of phospholipid synthesis in spermatozoa, fresh human spermatozoa were incubated with labeled fatty acids and 1‐acyl‐lysophosphatidyl choline (LPC) in the presence or absence of coenzyme A (CoASH). Both docosahexaenoic acid and palmitic acid were incorporated into phosphatidylcholine; however, this reaction was absolutely dependent upon the presence of CoASH in the incubation medium. The rate of incorporation of docosahexaenoic acid was 2.7‐fold higher than that of palmitic acid, but more palmitic acid was incorporated into phosphatidylcholine in the absence of LPC. These data provide direct evidence for acyl transferase activity in human spermatozoa and may furnish a mechanism for phospholipid remodeling in sperm membranes. The different incorporation rates of these fatty acids into phosphatidylcholine may be due to the kinetics of the activation step, long chain fatty acid:CoASH ligase (AMP), or the substrate specificity of the acyl transferase.
Balance of Apoptosis and Proliferation of Germ Cells Related to Spermatogenesis in Aged Men
Authors: Kimura Makoto , Itoh Naoki , Takagi Seiji , Sasao Takumi , Takahashi Atsushi , Masumori Naoya , Tsukamoto Taiji
Published: December 2015

ABSTRACT: To clarify whether germ cell apoptosis is related to a decrease of germ cells in the aged testis with impaired spermatogenesis, we investigated the apoptotic rate of each germ cell type. Testicular specimens were obtained by orchiectomy from 36 men with advanced prostate cancer and by testicular biopsy from 21 men with obstructive azoospermia, which served as controls. The terminal deoxynucleotidyl transferase‐mediated nick end labeling (TUNEL) technique was used to identify apoptosis. As a marker of cellproliferation activity, the expression of Ki‐67 was immunohistochemically evaluated. Expression of Bcl‐xl, which regulates apoptosis of germ cells, was also immunohistochemically examined. Histologically, except for spermatogonia, the ratios of primary spermatocytes, round spermatids, and elongated spermatids to Sertoli cells were significantly decreased in aged testes. The apoptotic rate in spermatogonia was significantly lower in aged men than it was in controls (0.11% ± 0.06% vs 0.34% ± 0.21%). Expression of Ki‐67 in spermatogonia was decreased in aged men (18.6% ± 6.0%) compared with that of controls (24.9% ± 3.3%), suggesting that germ cell proliferation diminished with aging. Consequently, the balance of spermatogonial proliferation and apoptosis showed no difference between the two groups. This was believed to be one of reasons why spermatogonial numbers in aged testes was similar to those of controls. The apoptotic rate of primary spermatocytes in aged men was significantly elevated compared with that of controls (0.60% ± 0.54% vs 0.22% ±0.12%), resulting in a decrease of the number of primary spermatocytes per Sertoli cell. The expression of Bcl‐xl was inversely correlated with the apoptotic rate in primary spermatocytes, suggesting that Bcl‐xl may be related to the regulation of primary spermatocyte apoptosis. Based on these findings, we conclude that accelerated apoptosis of primary spermatocytes might account for a part of the mechanism of germ cell loss in aging men.
Marked Differences in Protamine Content and P1/P2 Ratios in Sperm Cells From Percoll Fractions Between Patients and Controls
Authors: Mengual Lourdes , Ballescá José L. , Ascaso Carlos , Oliva Rafael
Published: December 2015

ABSTRACT: The various sperm cell types present in a simple ejaculate differ in their motility and morphology. However, little is known about the nuclear maturity of these sperm cells and their relationship with morphological and motile characteristics. Protamines are considered a good marker of sperm nuclear maturity since they are added to the DNA in the last stage of spermatogenesis. We have analyzed the P1/P2 ratio and the protamine content of subpopulations of human spermatozoa at different stages of maturation, isolated by density gradient centrifugation of ejaculated spermatozoa obtained from 3 groups of patients from our Assisted Reproduction Unit: 10 men of proven fertility, 12 oligozoospermic men, and 13 asthenozoospermic men. Four different fractions (F2‐F5) were collected from the top to the bottom of the Percoll gradient. Differences in the motion and morphology were found between the fractions in each of the groups studied, with fraction F5 being the one with the best morphology and motility. However, no significant differences in the P1/P2 ratio were found between fractions within the same group of samples, indicating that the P1/P2 ratio and the amount of protamines are relatively independent of the morphology and motility of sperm cells. In contrast, statistically significant differences were found in the P1/P2 ratio and in the relative amount of protamines between the 3 groups.
Progesterone Receptor Availability in Mouse Spermatozoa During Epididymal Transit and Capacitation: Ligand Blot Detection of Progesterone‐Binding Protein
Authors: Pietrobon Elisa Olivia , de, Los Monclus María , Alberdi Ángeles Antonio José , Fornés Miguel Walter
Published: December 2015

ABSTRACT: The goals of the present study were to determine the availability of progesterone (P4) receptor (P4r) in mouse sperm during maturation and capacitation and to make the first steps toward a characterization of P4r. It has been proposed that P4 is able to induce an acrosomal reaction (AR) by using a membrane P4r. This induction was verified in sperm isolated from the cauda epididymis (fully mature) when incubated in specific conditions that capacitate sperm. First, we set up the conditions in our laboratory to induce an AR in mature and capacitated sperm triggered by P4 that was detected by a chlortetracycline (CTC) assay. Then, we examined sperm isolated from the caput epididymis (immature) incubated under conditions that support cauda sperm capacitation and found that the AR could not be detected. Moreover, P4 was unable to induce the AR when it was applied to sperm isolated from either region and incubated under conditions that did not support capacitation. These results can be explained by changes in P4r availability. A suitable marker for P4r is the gold (Au)‐P4 complex. This marker shows a binding capacity that can be visualized directly by electron microscopy (EM) and indirectly by silver‐enhanced methods with light microscopy. The Au‐P4 complex was localized in capacitated cauda sperm at the dorsal edge of the head. Using these techniques, we observed a significant decrease in both noncapacitated cauda sperm and caput sperm (whether incubated in capacitating media or not). Genomic P4r could be responsible for the signal detected, but antibodies against the P4 nuclear receptor did not recognize any sites in the sperm by immunostaining methodology. Instead, a 44‐kd protein band was detected in the sperm by a ligand blot assay. In conclusion, P4 promotes the AR in capacitated cauda sperm but is unable to do so in noncapacitated or immature sperm because the availability of P4r increases during epididymal transit and after capacitation. The P4r responsible for this behavior is different from a classical nuclear receptor—on the basis of the immunostaining results—and is probably a protein close to 44 kd—on the basis of the ligand assay results.
Penile Venous Anatomy: An Additional Description and Its Clinical Implication
Authors: Hsu Geng‐Long , Hsieh Cheng‐Hsing , Wen Hsien‐Sheng , Chen Yi‐Chang , Chen Shyh‐Chyan , Mok Martin S.
Published: December 2015

ABSTRACT: The human penile venous system has been well studied and described but the demonstration of extra venous channels in imaging films prompted us to seek refinement of our anatomical knowledge of this venous system. Cavernosography in 37 patients who had venous stripping surgery and now suffered recurrent erectile dysfunction consistently showed an independent vein, smaller than the deep dorsal vein, running almost in the same position of the deep dorsal vein even though the latter had been removed unequivocally in previous surgery. Cavernosography in 9 patients who underwent intraoperative films also demonstrated the presence of this cavernosal vein in addition to the deep dorsal vein. Meticulous dissection of the penis under the microscope was then performed in 21 male cadavers and we found a cavernosal vein coursing along each corpus cavernosum all the way distally to the glans and draining directly into the Santorini's plexus in 19 subjects. This is in contrast to the previous description that this cavernosal vein was a short vein in the penile hilum. Two sets of para‐arterial veins, which have not been reported in the literature, were found to accompany each dorsal artery in all 21 subjects. This more extensive and extra venous drainage might have important implication for venous stripping surgery in the treatment of erectile dysfunction.
Zyxin, Axin, and Wiskott‐Aldrich Syndrome Protein Are Adaptors That Link the Cadherin/Catenin Protein Complex to the Cytoskeleton at Adherens Junctions in the Seminiferous Epithelium of the Rat Testis
Authors: Lee Nikki P. Y. , Mruk Dolores D. , Conway Anne M. , Cheng C. Yan
Published: December 2015

ABSTRACT: During spermatogenesis, the movement of germ cells across the seminiferous epithelium is associated with extensive junction restructuring. Yet the underlying mechanism (or mechanisms) that regulates these events is largely unknown. If the molecular architecture of the cell‐cell actin‐based adherens junction (AJ), such as ectoplasmic specialization (ES) and tubulobulbar complex— two testis‐specific AJ types, is known, many functional mechanistic studies can be designed. We thus undertook an investigation to study 3 adaptors in the seminiferous epithelium: zyxin, axin, and Wiskott‐Aldrich syndrome protein (WASP). All 3 adaptors were shown to be products of Sertoli and germ cells. Zyxin was shown to be a stage‐specific protein that was most prominent during stages V—VII and restricted mostly to pachytene spermatocytes, but it could also be detected at the site of basal and apical ectoplasmic specialization (ES). Zyxin, axin, and WASP were shown to be structurally linked to the N‐cadherin/β‐catenin/α‐actinin/actin complex but not to the nectin‐3/afadin or the β1‐ integrin—mediated protein complexes. Interestingly, zyxin, axin, and WASP are also structurally linked to vimentin (an intermediate filament protein) and α‐tubulin (the subunit of a microtubule), which suggests that they have a role (or roles) in the regulation of the dynamics of the desmosome‐like junction and microtubule. These results illustrate that zyxin, axin, and WASP are adaptors in both AJs and intermediate filament‐based desmosome‐like junctions. This raises the possibility that classic cadherins are also associated with vimentin‐based intermediate filaments via these adaptors in the testis. While virtually no N‐cadherin was found to associate with vimentin in the seminiferous tubules, it did associate with vimentin when testis lysates were used. Interestingly, about 5% of the E‐cadherin associated with vimentin in isolated seminiferous tubules, and about 50% of the E‐cadherin in the testis used vimentin as its attachment site. These data suggest that cadherins in the testis, unlike those in other epithelia, use different attachment sites to anchor the cadherin/catenin complex to the cytoskeleton. The levels of zyxin, axin, and WASP were also assessed during AF‐2364‐mediated AJ disruption of the testis, which illustrated a time‐dependent protein reduction that was similar to the trends observed in nectin‐3 and afadin but was the opposite of those observed for N‐cadherin and β‐catenin, which were induced. Collectively, these results illustrate that while these adaptors are structurally associated with the cadherin/catenin complex in the testis, they are regulated differently.
Interaction of PDC‐109, the Major Secretory Protein From Bull Seminal Vesicles, With Bovine Sperm Membrane Ca2+‐ATPase
Authors: Sánchez‐Luengo Silvia , Aumüller Gerhard , Albrecht Martin , Sen Parimal C. , Röhm Karlheinrich , Wilhelm Beate
Published: December 2015

ABSTRACT: PDC‐109 is the prevalent secretory protein from bovine seminal vesicles that binds to the midpiece of sperm once they pass the ampulla of the vas deferens during emission. Thereby, the protein changes biophysical membrane properties, eventually resulting in increased sperm motility. To elucidate the underlying biochemical mechanism, we have studied the ion‐pumping activity (Ca2+‐ATPase) in membrane preparations of bovine spermatozoa following in vitro incubation with the protein and analyzed whether PDC‐109 influences sperm motility. PDC‐109 was purified to homogeneity from bull seminal vesicle extracts using a newly described method. The effect of PDC‐109 on sperm motility was analyzed using the CASA‐method. These experiments clearly demonstrated that PDC‐109 significantly increases sperm motility. Calcium‐pumping mechanisms were analyzed by monitoring the effect of PDC‐109 on various parameters of enzyme activity of Ca2+‐ATPase in epididymal sperm plasma membranes and were compared with Ca2+‐ATPase activities from other organs and from epididymal sperm of different species, respectively. Specificity studies were performed using different Ca2+‐antagonists. Enzyme activities of both Mg2+‐dependent and Mg2+‐independent Ca2+‐ATPases increased in a dose‐dependent manner following the addition of the PDC‐109 (range 5–20 μg). Preincubation of PDC‐109 at temperatures above 37°C and pHs ranging from below 6.5 and above 8.5 led to the loss of the stimulatory effect. An analysis of enzyme kinetics pointed to irreversible, cooperative interaction of PDC‐109 with the enzyme. The effect was organ‐specific, that is, restricted to sperm ATPases, but it was not species‐specific, as it could be elicited also in rat sperm.
Immunofluorescence Reveals Ubiquitination of Retained Distal Cytoplasmic Droplets on Ejaculated Porcine Spermatozoa
Authors: Kuster Christopher E. , Hess Rex A. , Althouse Gary C.
Published: December 2015

ABSTRACT: The purpose of the present study was twofold: 1) to determine if antibodies raised against ubiquitin would recognize antigens associated with the porcine cytoplasmic droplet (CD), and 2) to determine if the same antibody would identify ubiquitinated substrates on the surface of morphologically abnormal boar spermatozoa. Permeabilization with the detergent Triton X‐100 (0.05%) showed virtually all CDs to be ubiquitin positive. Distal droplets (DDs) retained in situ on boar spermatozoa were readily labeled following Triton permeabilization, whereas DDs present on nonpermeabilized cells were not. Negative control preparations lacked the ubiquitin staining on the DD. The use of microtubes for fixation and incubation provided clearer images as well as better sperm cell distribution and density than an initial slide‐mounted technique. Immunoblotting indicated that larger amounts of ubiquitinated proteins were present in extracts from sperm cells from an ejaculate with an abnormally high percentage of retained DDs (52% DDs) compared to a morphologically normal sample (6% DDs). The primary antibody recognized both mono‐ubiquitin of bovine origin (8.5 kd) and human ubiquitin conjugate (35 kd), as demonstrated by Western blot. Preabsorption of the anti‐ubiquitin antibody with purified bovine ubiquitin was successful in preventing diaminobenzidine staining of sperm extract from the high DD ejaculate. The presence of antigens recognized by anti‐ubiquitin antibodies in the boar sperm CD, coupled with the possibility that superfluous ubiquitin species are detrimental to embryonic development by targeting critical paternally contributed zygotic organelles, raises concerns that retained DDs may be more detrimental to fertility than previously suspected.