bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence for Age‐Dependent Changes in Sertoli Cell Androgen Receptor Concentration
Authors: BUZEK S.W. , CASTON L.A. , SANBORN B.M.
Published: December 2015

Cytosol and nuclear receptor concentrations in Sertoli cells isolated from the testes of 15‐, 25‐, and 35‐day‐old rats were measured using hydroxylapatite separation procedures. In these cells the mean Kd of the cytosol receptor for methyltrienolone (3H‐R1881) ranged between 2.3 and 2.9 nM, and the concentration of cytosol androgen receptor per mg Sertoli cell DNA increased over the 15‐ to 35‐day age interval. However, when the data were expressed per mg cytosol protein, no increase was observed. The increase in receptor concentration per mg DNA paralleled the increase in cytosol protein/DNA ratio. The concentration of androgen receptor per mg DNA in nuclear extracts also increased with age. Consequently, total Sertoli cell androgen receptor increases over the time interval in which meiosis is first completed in the testis.
Reversible Harmless Interruption of Testicular Blood Supply in the Ram
Authors: VAN VLIET JOSKA , de RUITER‐BOOTSMA ANKE L. , OEI YOE H. , HOEKSTRA ANNE , de ROOIJ DIRK G. , WENSING CEES J.G.
Published: December 2015

An effective method of interrupting testicular blood flow temporarily and repeatedly in the ram has been developed. Blockade of flow has been achieved mechanically by an inflatable occluder placed around the testicular artery at the level of the spermatic cord. The effect of the blockade on total testicular blood supply was investigated using Doppler flowmetry and a percutaneous Xenon‐133 injection method. With both approaches, the blood flow changes after inflation or deflation of the occluders could be estimated satisfactorily. A substantial decrease of testicular blood flow was achieved in eight of the 10 testes with inflated occluders. However, there were indications that in the remaining two testes blockade of the arterial flow was not complete. After deflation of the occluders, blood flow was restored rapidly and completely in all testes. Macro‐ and microscopic examinations revealed no long‐term damage to the testis after blood flow interruptions lasting 30 or 60 minutes.
Suppression of Steroid Receptor‐Chromatin Interaction by Inhibitors Present in Ammonium Sulfate‐Fractionated Rat Testicular Androgen Receptor Preparations
Authors: TSAI YU‐HUI
Published: December 2015

Tritiated androgen receptor complex was partially purified by ammonium sulfate precipitation at 15 to 37% saturation from rat testicular or prostatic cytosols. A greater amount of prostatic tritiated androgen receptor complex bound to rat Sertoli cell chromatin than tritiated testicular androgen receptor complex. However, the testicular androgen receptor complex was more potent in competing for binding of either tritiated androgen receptor complex preparation to the Sertoli cell chromatin. Combining equal amounts of tritiated testicular and prostatic androgen receptor complexes resulted in lower binding than that observed with tritiated prostatic androgen receptor complex alone, while doubling the concentration of tritiated prostatic androgen receptor complex led to a 2‐fold increase in binding. It is conceivable that the partially purified testicular androgen receptor complex preparations contain factor(s) that inhibit the binding interaction of tritiated androgen receptor complex with chromatin, while there is no evidence for the presence of such inhibitors in tritiated prostatic androgen receptor complex preparations. Fractionation of testicular cytosol on DEAE‐cellulose yielded a flow‐through fraction with inhibitory activity and a bound fraction containing tritiated androgen receptor complex that was eluted with 0.3 M KCl and also showed inhibitory activity. These factors also inhibited the binding of tritiated estradiol receptor complex to chromatin. They were non‐dialyzable and their inhibitory effect was abolished after heating at 60 C for 30 minutes. It is concluded that ammonium sulfate‐fractionated rat testicular androgen receptor complex preparations contain inhibitors of steroid receptor‐chromatin interaction. The inhibitory factors can be fractionated into two distinct fractions by DEAE‐cellulose column chromatography. They are non‐dialyzable and heat‐labile. The precise chemical nature of the postulated inhibitor(s) remains to be determined.
Inhibition of Ornithine Decarboxylase Activity by Follicle Stimulating Hormone in Primary Culture of Rat Sertoli Cells
Authors: MADHUBALA RENTALA , SHUBHADA SANKARARAMAN , STEINBERGER ANNA , TSAI YU‐HUI
Published: December 2015

The effect of follicle stimulating hormone on the activity of ornithine decarboxylase (ODC) was determined in primary culture of rat Sertoli cells. Three different FSH preparations (NIH oFSH‐S‐15, S‐16, and eFSH) inhibited ODC activity in rat Sertoli cells under different media conditions. The inhibition was both time‐ and dose‐dependent. The mechanism of the FSH inhibitory effect was studied using dibutyryl cyclic adenosine monophosphate (dbcAMP), 1‐methyl‐3‐isobutylxanthine (MIX), forskolin, and isoproterenol. All of these agents, known to elevate cellular cAMP levels, inhibited ODC activity in cultured rat Sertoli cells. The combined effect of each of these substances plus FSH was either greater than, or equal to, that of FSH alone, and was not additive. Dibutyryl cyclic guanosine monophosphate had no effect on the ODC activity. These findings suggest that FSH inhibition of ODC activity in the rat Sertoli cell may be mediated by cAMP.
Characterization of β‐Adrenergic Receptors in Dispersed Rat Testicular Interstitial Cells
Authors: POYET P. , LABRIE F.
Published: December 2015

Recent studies have shown that β‐adrenergic agents stimulate steroidogenesis and cyclic AMP formation in mouse Leydig cells in culture. To obtain information about the possible presence and the characteristics of a β‐adrenergic receptor in rat testicular interstitial cells, the potent β‐adrenergic antagonist [125I]cyanopindolol (CYP) was used as ligand. Interstitial cells prepared by collagenase dispersion from rat testis were incubated with the ligand for 2 h at room temperature. [125I]cyanopindolol binds to a single class of high affinity sites at an apparent KD value of 15 pM. A number of sites of 6,600 sites/cell is measured when 0.1 μM (‐) propranolol is used to determine non‐specific binding. The order of potency of a series of agonists competing for [125I]cyanopindolol binding is consistent with the interaction of a β2‐subtype receptor: zinterol > (‐) isoproterenol > (‐) epinephrine = salbutamol >> (‐) norepinephrine. In addition, it was observed that the potency of a large series of specific β1 and β2 synthetic compounds for displacing [125I]cyanopindolol in rat interstitial cells is similar to the potency observed for these compounds in a typical β2‐adrenergic tissue, the rat lung. For example, the potency of zinterol, a specific β2‐adrenergic agonist, is 10 times higher in interstitial cells and lung than in rat heart, a typical β1‐adrenergic tissue. Inversely, practolol, a typical β1‐antagonist, is about 50 times more potent in rat heart than in interstitial cells and lung. The binding of [125I]cyanopindolol to interstitial cells shows a marked stereoselectivity, the agonist (‐) isoproterenol and the antagonist (‐) propranolol being 500 and 300 times more potent than their respective (+) enantiomers. The high selectivity and density of β2‐adrenergic receptors in rat testicular interstitial cells suggest a role of circulating and/or locally secreted physiologic epinephrine in the control of testicular function.
Improvement in the Quality and Fertilization Potential of a Human Sperm Population Using the Rise Technique
Authors: RUSSELL LONNIE D. , ROGERS B. JANE
Published: December 2015

Semen from 63 individuals participating in an in vitro fertilization program was processed using a modified rise technique. Overall normal morphology was significantly improved in the rise (79.2%) as compared with the unprocessed sample (57.8%), and six of seven specific morphologic abnormalities were significantly reduced. Motility was significantly enhanced from 51.8% in the unprocessed samples to 89.1% in the rise samples. Spermatozoa recovered in the rise portion of the sample represented 5.9% of the total sample. Ultrastructural morphometry revealed that the rise was relatively free of abnormal sperm forms, acellular debris and non‐sperm cellular elements as compared with the non‐rise portion of the sample or a typical unprocessed sample. Volume density measurements demonstrated that only 18.1% of the volume of the non‐rise sample was composed of normal spermatozoa compared with 83.4% of the volume of the rise. In a separate set of experiments utilizing 21 samples, the penetration of hamster eggs was significantly enhanced from 37.9% to 67.2% using spermatozoa from the initial washed sample and those from the rise, respectively. These data demonstrate the qualitative and quantitative improvements, as well as the increase in fertilizing potential, of the rise portion of the semen sample.
Nitrocellulose and Polyvinyl Coatings Prevent Sperm Adhesion to Glass Without Affecting the Motility of Intact and Demembranated Human Spermatozoa
Authors: CHAPEAU CHRISTINE , GAGNON Dr.CLAUDE
Published: December 2015

The effectiveness of several glass coating agents in preventing sperm adherence to glass surfaces for both intact and demembranated human spermatozoa was investigated. These agents included bovine serum albumin (BSA), Sigmacote, poly glulys, Collodion and Formvar. The presence of at least 5% of seminal plasma in sperm suspensions prevented sperm adhesion to glass surfaces. On the other hand, 56% of washed spermatozoa resuspended in an isoosmotic buffer containing 1 mg/ml of BSA attached to glass. Collodion, Formvar and Sigmacote reduced sperm attachment to glass to 2, 5 and 7%, respectively. BSA was partially effective, with 20% sperm adherence to glass, and poly glu‐lys was totally ineffective. Whereas Sigmacote and BSA coatings lacked transparency, Collodion always achieved the best light transmission. Demembranated reactivated spermatozoa were all attached to glass within 90 seconds of contact. This adhesion was prevented by Collodion and Formvar. Other agents were less effective or interfered with motility. In contrast to intact spermatozoa, demembranated spermatozoa have a very low progressiveness ratio (vector speed/track speed), a wide beating amplitude and because of their whiplash flagellar movement, their motility resembles that of hamster capacitated spermatozoa.
Cryosurvival of Human Spermatozoa Frozen in Eight Different Buffer Systems
Authors: WEIDEL LOUISE , PRINS GAIL S.
Published: December 2015

The present study was conducted to ascertain optimal cryoconditions for human spermatozoa by 1) comparing the relative cryoprotective efficiency of eight buffer systems and 2) assessing various cryovials and thaw rates under two freeze rates. Spermatozoa that were cryopreserved in one of four zwitterion buffers (TES‐Triscitrate‐egg yolk‐glycerol; TES‐Tris‐citrate‐I) maintained higher progressive motility at 0, 1, 2, and 4 hours post‐thaw as compared to cells frozen in glycerol only, citrate‐egg yolk‐glycerol and TES‐Tris‐citrate‐egg yolk without glycerol (TES‐Tris‐citrate‐III; P < 0.01). Freezing in TES‐Tris‐citrate‐I also resulted in spermatozoa that penetrated the furthest distance through cervical mucus and possessed the highest percent live spermatozoa when compared to other cryoprotective media. Spermatozoa were analyzed for their ability to penetrate zona‐free hamster ova and no difference was found between buffers when the assay was corrected for progressive motility. After removal of seminal plasma/buffers and incubation for 2 hours in BWW, TES‐Tris‐citrate‐II and TES‐Tris‐citrate‐milk showed the greatest sperm longevity (P < 0.05). Pooled semen was extended in TES‐Tris‐citrate‐I and frozen in straws or ampoules in static N2 vapor or in pellets on dry ice. Thaw bath temperatures ranged from 0 to 37 C. Post‐thaw progressive motility and cervical mucus penetration were similar in all treatment groups. In conclusion, the present results indicate the use of TES‐Tris‐citrate‐I for cryopreservation of human spermatozoa. With this optimal cryoprotective buffer, the containers and thaw rates used have little effect on human sperm cryosurvival.
Correlation Between In Vitro Fertilization and Human Sperm Density and Motility
Authors: GERRIS JAN , KHAN IQBAL
Published: December 2015

The conventional sperm characteristics of density (millions per milliliter) and motility, scored in a semi‐subjective way, were correlated with results of an on‐going in vitro fertilization and embryo transfer program. No male infertility patients were included in this study. Individual characteristics of the “successful” ejaculates are described. Sperm densities in the original ejaculate of more than 10 × 106 spermatozoa/ml did not significantly improve outcome (P < 0.01). In contrast, sperm motility seemed to play the most important role, since most pregnancies (12/14) occurred using sperm samples with ≥ 60% total motility (P < 0.001). The incidence of multipronuclear fertilization is also described and discussed. These data, which were collected during 1984 in the in vitro fertilization unit of Professor R. Schoysman and coworkers (Vilvoorde, Brussels), may help to make fertilization in vitro and embryo transfer a viable method in cases of mild male subfertility, and to provide guidance in preparing some couples for the combined use of husband and donor semen if a sufficient number of oocytes are obtained.
Automatic Analysis of Human Sperm Motion
Authors: KATZ D. F. , DAVIS R. O.
Published: December 2015

A new computerized methodology is described in which sperm movement characteristics are analyzed automatically. Video fields containing images of spermatozoa are electronically digitized, and the centroid position of each sperm head is determined. Over time, strings of centroids identify the swimming paths of individual spermatozoa. Details of path acquisition are described for human spermatozoa in semen, and include a discussion of how individual cells are identified and distinguished from each other. A diversified set of movement characteristics is computed for each spermatozoon, including two new measures of path shape based on the instantaneous turning angle. The traditional and the new measures of vigor and swimming pattern are evaluated and compared for consistency and redundancy. Analysis of data from human semen indicates that the new angular measures may be particularly useful in discriminating between spermatozoa exhibiting widely different patterns of motion.
Variation in the Effect of a Nongonadotropic Leydig Cell Stimulating Factor in Testicular Interstitial Fluid After Exposure of the Testis to a Single Episode of Heat Treatment
Authors: ISHIDA HAJIME , RISBRIDGER GAIL P. , DE KRETSER DAVID M.
Published: December 2015

The activity of a nongonadotropic factor present in rat testicular interstitial fluid and capable of stimulating Leydig cell testosterone production was measured at intervals for 8 weeks after exposure of the testis to a single episode of heat treatment (43 C for 15 minutes). The activity of this factor was determined using a Leydig cell bioassay and the testicular albumin space (a measure of the interstitial fluid volume) was determined after the injection of 125I‐labeled bovine serum albumin. Additionally, the response of interstitial cells to hCG stimulation was measured at each time point after heat treatment. The response of interstitial cells to hCG and the activity of the interstitial fluid were both increased transiently 2 or 3 weeks after heat treatment coincident with the disruption of spermatogenesis. These data provide further evidence for the local interaction of the tubules and interstitium and imply a role for the testicular interstitial fluid factor in these local regulatory mechanisms.
Quantification of Bovine Sperm Separation by a Swim‐up Method Relationship to Sperm Motility, Integrity of Acrosomes, Sperm Migration in Polyacrylamide Gel and Fertility
Authors: PARRISH JOHN J. , FOOTE ROBERT H.
Published: December 2015

The number of bovine spermatozoa separated in a swim‐up procedure was quantified using an electronic cell counter. In an initial test of the swim‐up procedure, non‐frozen sperm samples with different ratios of live to dead cells were prepared and tested for the number of spermatozoa counted by the swim‐up procedure. In ejaculates from six bulls, the number of spermatozoa swimming up was related to the number of live cells present (R2=0.97). Next, sperm quality of frozen‐thawed semen immediately after thawing was measured at 37 C by swim‐up sperm count, sperm motility, spermatozoa with an intact acrosome and migration in Polyacrylamide gel and then compared with the fertility of the semen used for artificial insemination. Twenty‐nine ejaculates of frozen‐thawed semen from 11 bulls were evaluated. Correlations with fertility were highest on an ejaculate basis for motility (r = 0.41, P = 0.05) and for swim‐up sperm count (r = 0.35, P = 0.06). On a bull basis, swim‐up sperm count had the highest correlation with fertility (r = 0.59, P = 0.06). In a multiple regression model to predict male fertility that included all described measures of semen quality, a R2 value of 0.69 was obtained. This is the first report showing that the ability of spermatozoa to swim out of a more dense medium (whole milk‐glycerol extender) into culture media is quantitatively related to in vivo fertility.
Serum Testosterone Response to Desialylated Human Choriogonadotropin in Man
Authors: LIU LINDA , WEHMANN ROBERT E. , ROSA CESAR , BIRKEN STEVEN , NISULA BRUCE C.
Published: December 2015

To assess the in vivo steroidogenic activity of desialylated human choriogonadotropin (hCG) in man, highly purified desialylated hCG was administered as a constant intravenous infusion over 6 hours to four normal men at a rate sufficient to maintain substantial levels of desialylated hCG in the serum. The mean percent change of serum testosterone from baseline during the first 6 hours in men given an infusion of desialylated hCG was compared to that in saline‐infused controls and that in men given highly purified intact hCG. The mean change of serum testosterone at 6 hours in the group infused with desialylated hCG (129% of baseline) was significantly greater than that of the controls (69% of baseline). Furthermore, the response to desialylated hCG could not be distinguished from that of hCG (125% of baseline). It was concluded that desialylated hCG, when given as a constant intravenous infusion, can elicit a substantial serum testosterone response comparable to that seen with purified hCG, and thus, that desialylated hCG behaves as an agonist of the LH/CG receptors on human Leydig cells.
The Sperm Chromatin Structure Assay Relationship with Alternate Tests of Semen Quality and Heterospermic Performance of Bulls
Authors: BALLACHEY BRENDA E. , EVENSON DONALD P. , SAACKE RICHARD G.
Published: December 2015

Data obtained by the sperm chromatin structure assay (SCSA) on spermatozoa from nine bulls were correlated with fertility, measured by heterospermic performance (–0.94, P < 0.01) and by alternate tests of sperm quality, including motility, acrosome integrity, Sephadex filtration and morphology of spermatozoa (all significant at P < 0.05 to P < 0.01). The SCSA uses flow cytometry to determine the susceptibility of nuclear DNA to low pH‐induced denaturation in situ as measured by the ratio of acridine orange binding to double‐ or single‐stranded DNA. The error associated with multiple SCSA measurements was relatively low. The primary finding is that the assay of chromatin structure stability performed on killed spermatozoa was as highly correlated with the heterospermic performance of semen as the best of the classical tests for semen quality. The SCSA may therefore be a highly useful technique for evaluation of sperm quality.
HCG Treatment Increases Intratesticular Pressure in the Abdominal Testis of Unilaterally Cryptorchid Rats
Authors: HJERTKVIST MATS , BERGH ANDERS , DAMBER JAN‐ERIK
Published: December 2015

Adult, unilaterally cryptorchid rats were given a single subcutaneous injection of hCG. HCG treatment of 100 I.U. (but not 10 I.U.) resulted in a marked increase in intratesticular pressure (approximately 40 mm Hg) in the abdominal testis that was maximal 24 hours after treatment. This increase in pressure is caused by increased vascular permeability coupled with insufficient lymph drainage. In the scrotal testis, hCG treatment resulted in increased vascular permeability and lymph flow, but this did not result in a marked increase in testicular pressure. No morphologic signs of hCG‐induced damage were observed in either the abdominal or scrotal testis 10 days after hCG treatment. Testicular microcirculation, as studied by laser doppler flowmetry, was abnormal in the abdominal testis, but hCG treatment inhibited vasomotion in both the abdominal and scrotal testis.
Effect of Repeated Washing on Sperm‐Bound Immunoglobulin G
Authors: HAAS GILBERT G. , D'CRUZ OSMOND J. , DENUM BARRETT M.
Published: December 2015

The effect of sperm washing on the stability of sperm‐bound immunoglobulin G (IgG) antibody derived from plasma from four patients and also IgG bound in vivo on the spermatozoa of four other men was quantitatively evaluated. In the first series of experiments, human spermatozoa were incubated with an IgG antibody‐containing plasma and subjected to 18 cycles of sperm washing. In the second set of experiments, spermatozoa from men positive for sperm‐bound IgG were subjected to four cycles of sperm washing. The amount of residual antibody bound to a constant number of spermatozoa was quantitated by a radiolabeled antiglobulin assay during and following the washing procedures. There was no significant loss of sperm‐bound antibody due to the washing procedures. The results of these studies undermine the utility of sperm washing as an effective treatment of antibody‐mediated infertility in men.
Pituitary‐Testicular Reserve in Men with Low Serum Testosterone and Normal Serum Luteinizing Hormone
Authors: GLASS ALLAN R.
Published: December 2015

Men with low serum testosterone levels who do not have elevated serum LH levels are generally thought to have hypothalamic‐pituitary dysfunction. To evaluate this concept, seven men with a combination of low serum testosterone and normal serum LH underwent standard tests of hypothalamic‐pituitary‐testicular reserve. Pituitary reserve, tested with LHRH, showed exaggerated responses in two subjects, low‐normal responses in one subject, and normal responses in the remaining four. Testing of hypothalamic‐pituitary reserve with clomiphene showed normal gonadotropin responses in six subjects and blunted response in one (the same subject with the low LHRH response). Direct stimulation with hCG showed normal percentage increases in testosterone but low absolute levels, comparable to responses in patients with Klinefelter's syndrome. However, 17‐OH‐progesterone responses to hCG were lower in these subjects than in either controls or subjects with Klinefelter's syndrome. During follow‐up, one subject developed frank primary testicular failure. It was concluded that men with low serum testosterone but normal serum LH are a heterogeneous group, and this pattern occasionally reflects early primary testicular failure rather than hypothalamic‐pituitary dysfunction. Standard tests of pituitary‐testicular reserve are generally not useful in defining abnormal hormonal output, although measurement of the 17‐OH‐progesterone response to hCG may improve their diagnostic utility.
Localization of the Human Prostatic Secretory Protein PSP94 and its mRNA in the Epithelial Cells of the Prostate
Authors: BRAR ANOOP , MBIKAY MAJAMBU , SIROIS FRANCINE , FOURNIER SYLVIE , SEIDAH NABIL G. , CHRÉTIEN MICHEL
Published: December 2015

The histologic distribution of a prostatic secretory protein of 94 amino acids (PSP94) and its mRNA were examined simultaneously by conventional immunohistochemistry and in situ hybridization histochemistry. The results show localization of immunoreactive PSP94 and its mRNA in the epithelial cells of the prostate gland, thus providing strong evidence of PSP94 synthesis in these cells. Preliminary analysis of PSP94 by radioimmunoassay and of its mRNA by Northern blot analysis indicates that PSP94 biosynthesis in pathologic prostatic tissues is variable. The value of PSP94 as a marker of prostate gland secretory activity is discussed.
Differential Effects of (+) and (−)Gossypol Enantiomers on Mitochondrial Function and Proliferation of Cultured TM4 Cells
Authors: TANPHAICHITR NONGNUJ , FITZGERALD LISA M. , MATLIN STEPHEN A.
Published: December 2015

The in vitro effects of (+) and (–)gossypol enantiomers on thde mitochondrial functions of mouse transformed Sertoli, TM4 cells were investigated by monitoring mitochondrial rhodamine 123 accumulation. When TM4 cells were cultured in medium without fetal calf serum, 5 μg/ml of both enantiomers caused similar declines in mitochondrial rhodamine 123 staining. By contrast, (–)gossypol had a greater adverse action than did (+)gossypol on the mitochondria of TM4 cells that were cultured in medium supplemented with 2% fetal calf serum. Construction of dose response curves for the effects of the two enantiomers on rhodamine 123 accumulation by TM4 cells after 5 hr of drug treatment gave an EC50 of 7.5 μg/ml for the (–)gossypol isomer compared with 18 μg/ml for the (+)gossypol isomer. Similarly, TM4 cell proliferation was also more disturbed by (–)gossypol in medium supplemented with other concentrations of fetal calf serum. The results suggest that the lower effect of (+)gossypol on TM4 cells may be attributed to its higher affinity to serum components, which may impede its entrance into TM4 cells.
Origin and Fate of Autophagosomes in Leydig Cells of Normal Adult Rats
Authors: TANG X. M. , CLERMONT Y. , HERMO L.
Published: December 2015

Autophagosomes were observed frequently in electron microscope photographs of Leydig cells from normal adult rat testis. Their formation, evolution and fate were analyzed morphologically in preparations treated to show cytidine monophosphatase (CMPase) and glucose‐6‐phosphatase (G‐6‐Pase) activities and in animals sacrificed at various time intervals ranging from 5 min to 6 hrs after a single intratesticular injection of cationic ferritin. Analysis of the morphologic data led to the following interpretation and model. Preautophagosomal structures appeared as flattened, elongated membranous profiles. These expanded, took on a C‐shape and fused at their edges to demarcate a small cytoplasmic territory containing normal‐looking smooth endoplasmic reticulum (ER) and mitochondria. Such early autophagosomes were thus delimited by two membranes separated by a narrow lumen. Following fusion of these elements with secondary lysosomes, the space between the two membranes increased in size, the inner membrane disintegrated and the enclosed organelles no longer could be identified. The late autophagosomes then reached the cell surface and appeared to exocytose their residual content.

In contrast to secondary lysosomes and trans‐Golgi elements, which were CMPase‐positive, the preautophagosomal flattened membranous elements and early autophagosomes were CMPase‐negative. The late autophagosomes on the contrary were CMPase‐positive. While ER cisternae were G‐6‐Pase‐positive, the pre‐, early and late autophagosomal structures were unreactive for this enzyme. Cationic ferritin tracer experiments showed that only late autophagosomes became labeled with cationic ferritin following their fusion with secondary lysosomes into which the tracer had accumulated following its endocytosis from the cell surface. These various data suggested that the membrane‐delimiting autophagosomes did not derive from the ER or the Golgi apparatus and that autophagocytic structures seemingly acquired their hydrolytic enzymes by fusion with secondary lysosomes of endocytic origin.
Pituitary Gonadotropin Content in Gonadectomized Rats: Immunoassay Measurements Influenced by Extraction Solvent and Testosterone Replacement
Authors: SPITZBARTH TEALA L. , HORTON TERESA H. , LIFKA JAMI , SCHWARTZ NEENA B.
Published: December 2015

Increasing interest in events regulating gonadotropin synthesis and secretion led to the reexamination, by radioimmunoassay (RIA), of the pituitary content of luteinizing hormone (LH) and follicle stimulating hormone (FSH). In a preliminary experiment, the importance of the composition of the solvent used to homogenize pituitaries on the detection of LH and FSH was demonstrated by RIA. Experiment 1 demonstrated that a drop in pituitary FSH occurred acutely following gonadectomy in males, but not in females. Experiment 2 tested whether this decline resulted from loss of testosterone or inhibin. Males were castrated and treated daily with oil or testosterone propionate, porcine follicular fluid, porcine serum, or testosterone plus follicular fluid for 2 to 14 days. Castration lowered pituitary FSH at 2, 4, and 6 days. Follicular fluid suppressed serum FSH, but not LH, and did not prevent the fall in pituitary FSH. Testosterone blocked the rise of serum LH and FSH and prevented the decline in pituitary FSH.
Diagnostic Value of Sperm Function Tests and Routine Semen Analyses in Fertile and Infertile Men
Authors: WANG CHRISTINA , CHAN STEVEN Y. W. , NG MATTHEW , SO WILLIAM W. K. , TSOI WAI‐LOONG , LO TORRY , LEUNG A.
Published: December 2015

The results of routine semen analyses, the zona‐free hamster oocyte penetration test, the hypoosmotic swelling test, and semen adenosine triphosphate levels were studied in 66 fertile and 130 infertile men. Multivariate discriminant analysis demonstrated that routine semen parameters including semen volume, sperm count, percent sperm motility, and percent normal spermatozoa in combination could predict the fertility of these patients with 70.4% accuracy. Of the three sperm function tests evaluated, the zona‐free hamster oocyte penetration test and the hypoosmotic swelling test were selected by the multivariate discriminant analysis as variables capable of providing significant information on the fertility status of the patients. However, the addition of the results of these two tests to the routine semen analysis did not significantly improve the predictability of fertility. The overall correct prediction rate was 77.6% after incorporation of the results of these two sperm function tests. In this group of subjects, the presently available sperm function tests did not predict the fertility status of a patient with a high degree of accuracy.
Transferrin Secretion in Response to Different Modes of FSH Stimulation and Cycloheximide in Superfused Sertoli Cell Cultures
Authors: JAKUBOWIAK ANDRZEJ , JANECKI ANDRZEJ , STEINBERGER ANNA
Published: December 2015

The influence of different modes of FSH stimulation and cycloheximide on transferrin secretion by rat Sertoli cells was investigated using a superfusion culture system. Sertoli cells from 18‐day‐old rats were cultured in serum‐free medium on Matrigel‐covered slides first in static conditions for 19 hours, and then superfused at a flow rate of 2.5 ml/hour. After an equilibration period of 48 hours to establish the basal rate of transferrin secretion, the cultures were exposed to various modes of FSH stimulation. Sertoli cells stimulated intermittently (20 min/2 hours) up to 22 hours responded to each consecutive FSH pulse with a rapid increase of transferrin secretion followed by a decline toward basal values. Continuous 22‐hour exposure to FSH elicited an immediate increase followed by irregular fluctuations and a transient decline towards the baseline. With either mode of FSH stimulation, there was a secondary prolonged increase in transferrin secretion. Although cultures stimulated intermittently or continuously during the entire experimental period (22 hours) secreted similar cumulative amounts of transferrin (10.8 ± 0.5 μg and 11.1 ± 0.8 μg, respectively), there was a direct correlation between the secreted amount of transferrin and the duration of FSH exposure up to 8 hours. Addition of cycloheximide decreased both basal and FSH‐stimulated transferrin secretion. However, even when cycloheximide was added 1 hour before FSH, an early secretory peak in response to FSH was still observed. It was concluded that, regardless of the mode of FSH stimulation, transferrin secretion shows a biphasic response in which an initial increase within minutes of FSH exposure is followed by a return to basal levels, and then a delayed and prolonged secondary increase. Secondly, cumulative amounts of transferrin can vary with the mode of FSH stimulation; finally, Sertoli cells appear to secrete mainly newly synthesized transferrin, but a small storage pool may also exist for immediate release by FSH independently of new protein synthesis.
The Correlation Between the Gossypol Contents in Blood Plasma, Rete Testis Fluid, and Cauda Epididymal Fluid Following Chronic Treatment with Gossypol in Rats
Authors: WANG JUMING , QIU JIANPING , WU XIAOLAN , ZHANG ZHAORONG , SHAO YU , CAO RUQIN
Published: December 2015

Concentrations of gossypol in blood plasma, rete testis fluid, and fluid from the caudia epididymidis were measured simultaneously by high performance liquid chromatography in rats treated with gossypol (15 mg/kg daily for 3 weeks). Antispermatogenic effects were demonstrated by loss of sperm motility in the cauda epididymidis and structural changes in the testis. It was found in these treated rats that concentrations of gossypol were lower in rete testis fluid compared with blood plasma but increased significantly in fluid from the cauda epididymidis. The results indicate a restriction of the blood‐testis barrier to gossypol and its local concentration in the epididymis after fluid resorption.
Induction of Varicocele in the Dog: I. Partial Ligation of the Left Renal Vein Does Not Induce a Varicocele in the Dog
Authors: LAVEN JOOP S. E. , WENSING CEES J. G.
Published: December 2015

Induction of varicocele was attempted by partial ligation of the left renal vein in 10 male dogs. The effects on sperm count, sperm motility, and sperm morphology, as well as on hemodynamics, were assessed. Furthermore, testicular, vascular, and kidney morphology was studied. Changes in the diameter and consistency of the left spermatic cord were found to be temporary. Total sperm count, sperm motility, and the total number of oval forms were not significantly altered. Hemodynamic studies revealed a renocaval pressure gradient, but retrograde flow in the distal part of the left testicular vein could not be observed by arteriography. A collateral network was found to compensate for the restricted left renal vein. Histologic examination revealed no damage to the seminiferous epithelium. Changes were not found in the kidney and left pampiniform plexus. Although some temporary changes induced by the partial ligation of the left renal vein are suggestive of varicocele, this hemodynamic study shows that the presented dog model does not mimic varicocele as encountered in man.
Genetically Defined Mouse Models of Male Infertility
Authors: CHUBB CURTIS
Published: December 2015

The review has four objectives. The first objective is to summarize our studies of gene mutations that induce infertility in male mice. The second objective is to stress the power of mouse genetics and its application to male reproductive biology. The third objective is to provide useful references to the literature about gene mutations affecting male mouse fertility. Our goal is to cite references that can be used as a starting point for reading, not to present a comprehensive list of references. The fourth objective is to present a summary of selected data collected from mice representing 21 mouse strains.
Evaluation of Computerized Analysis of Sperm Movement Characteristics and Differential Sperm Tail Swelling Patterns in Predicting Human Sperm In Vitro Fertilizing Capacity
Authors: CHAN S.Y.W. , WANG C. , NG M. , TAM G. , LO T. , TSOI W.L. , NIE G. , LEUNG J.
Published: December 2015

Multivariate discriminant analysis was used to evaluate the usefulness of computer image analysis of sperm movement characteristics and differential patterns of sperm tail swelling after hypoosmotic treatment for predicting the human sperm in vitro fertilizing capacity assessed by the zona‐free hamster egg penetration assay. Fifty‐five semen samples, mostly normospermic, from untreated infertility clinic patients were analyzed. The % normal sperm morphology, linearity of seminal sperm movement, seminal sperm head beat frequency, mean and maximum amplitudes of lateral head displacement, and hypoosmotic sperm tail swelling patterns c, d and f were selected by multivariate discriminant analysis to be capable of discriminating the samples exhibiting the presence or the absence of sperm in vitro fertilizing capacity. The % total sperm tail swelling did not give additional information about in vitro fertilizing capacity. These preliminary data suggest that computer image analysis of sperm movement characteristics and differential evaluation of hypoosmotic sperm tail swelling might be useful for the prediction of human sperm fertility. Further prospective studies are necessary to validate their predictive functions.
The Changing Profiles of L‐Ornithine Decarboxylase and S‐Adenosyl‐L‐Methionine Decarboxylase Activities in Testicular Cell Types During Sexual Maturation of Male Rats
Authors: SHUBHADA SANKARARAMAN , DAVER REKHA , TSAI YU‐HUI
Published: December 2015

To understand the involvement of polyamines in testicular maturation and spermatogenesis, the activity of two enzymes involved in the polyamine synthetic pathway, L‐ornithine decarboxylase (ODC) and S‐adenosyl‐L‐methionine decarboxylase (AdoMetDC) were determined in whole testis and isolated testicular cell types during the sexual maturation of male rats. ODC activities per mg protein in whole testis and cultured Sertoli cells from 12‐ to 14‐day‐old rats were very high but declined as the age of the animals increased. ODC activities in both germ cells and interstitial cells increased as the age of the rats increased, reached a maximum at 22 days and then declined rapidly. On the other hand, AdoMetDC activity in whole testis, Sertoli cells, germ cells and interstitial cells was highest from 27 to 35 days, 18 to 22 days, 22 to 27 or 35 days, and 27 days of age, respectively, then decreased and remained at a constant lower level after 45 days. After 35 or 45 days of age, cellular AdoMetDC activities were relatively higher than ODC activities in the whole testis and germ cells, while the activity of both enzymes was comparable in the Sertoli cells. Correlation of these data with the development of germ cells during rat testicular maturation suggests that the high ODC activity in both Sertoli and germ cells is associated mainly with cell proliferation while the increase in AdoMetDC activity is most likely associated with the maturation of Sertoli cells and the meiosis of germ cells, particularly the formation of pachytene spermatocytes and the meiotic cell division of spermatocytes into spermatids.
Synchronous Assay for Human Sperm Capacitation and the Acrosome Reaction
Authors: de JONGE C.J. , MACK S.R. , ZANEVELD L.J.D.
Published: December 2015

A synchronous acrosome reaction system was established for human spermatozoa. Seminal plasma is removed from the spermatozoa by centrifugation and the washed spermatozoa are capacitated in a modified BWW medium (without exogenous substrates) containing 35 mg/ml human serum albumin for 3 h at 37 C. Subsequently, 10 μM ionophore A23187 (final concentration) is added, the mixture incubated for 15 min at 37 C and the percent acrosome reaction determined by a modified triple stain technique (trypan blue stain omitted). Since no significant decrease in sperm motility occurs during incubation or after ionophore treatment, a vital stain does not need to be employed, allowing the use of any acrosome detection technique. The average percentage of acrosome‐rejacted spermatozoa after ionophore treatment (40 ± 10%) was about 2‐ to 3‐fold higher than that seen without ionophore treatment. Ionophore treatment for 15 min failed to stimulate the acrosome reaction in spermatozoa incubated for less than 3 h. Additionally, the presence of substrates in the BWW medium, higher sperm numbers, increased ionophore concentrations or longer incubation periods did not enhance the induction of the acrosome reaction. Ionomycin, a more specific calcium ionophore than A23187, produced essentially the same results as A23187 but tended to decrease sperm motility. This synchronous acrosome reaction system for human spermatozoa is relatively simple and can be used to study the effect of modulators on capacitation and/or the acrosome reaction.
Treatment of Men with Idiopathic Oligozoospermic Infertility Using the Aromatase Inhibitor, Testolactone Results of a Double‐Blinded, Randomized, Placebo‐Controlled Trial with Crossover
Authors: CLARK RICHARD V. , SHERINS RICHARD J.
Published: December 2015

The hypothesis that increased estradiol production may be the cause of impaired spermatogenesis in infertile men with idiopathic oligozoospermia was tested by administering the aromatase inhibitor, testolactone, and by assessing its effects on sperm output and fertility. Our study was a randomized, placebo‐controlled doubleblind crossover trial. Subjects (n = 25) with infertility due to unexplained oligozoospermia were given testolactone (2 g/day) or placebo for 8 months followed by crossover to the other treatment for an additional 8 months. Total estradiol and testosterone levels during testolactone exposure did not change from basal and placebo values. However, sex hormone‐binding globulin binding capacity consistently decreased (30%, p < 0.01) and free testosterone levels increased (36%, p < 0.01). Free estradiol values increased but not significantly. Additionally, LH and FSH serum levels increased by 15% and 20%, respectively (p < 0.05), and 17α‐hydroxyprogesterone values increased by 90% (p < 0.05) during drug administration. Sperm output and semen quality remained unchanged during either testolactone or placebo treatment, and no pregnancies occurred during the 16‐month study. These data suggest that chronic administration of testolactone at this dose fails to maintain aromatase inhibition despite depression of 17,20‐desmolase activity with elevated 17α‐hydroxyprogesterone and depressed SHBG binding capacity with elevation of free testosterone. Testolactone is not efficacious in the treatment of idiopathic oligozoospermic infertility.
Secretion of Plasminogen Activator in Response to Follicle‐Stimulating Hormone in Culture Medium of Human Testicular Cells from Biopsy Specimens
Authors: NISHIMUNE YOSHITAKE , SAWADA KEN , TAJIMA YOUICHI , WATANABE DAISUKE , MAEKAWA MAMIKO , SAKAMAKI KAZUHIRO , OKUYAMA AKIHIKO , NAKAMURA MASAHIRO , NAMIKI MIKIO , MATSUMOTO KEISHI , MATSUO OSAMU
Published: December 2015

Testicular cells from human biopsy specimens were cultured and plasminogen activator secreted into the medium was investigated. Follicle stimulating hormone stimulated plasminogen activator secretion. SDS‐polyacrylamide gel electrophoresis followed by zymography showed that the molecular weight of plasminogen activator produced by human testicular cells is the same as that of human urokinase. Antigenicity was also found to be of the urokinase type but not of the tissue type.
Possible Role of Elongated Spermatids in Control of Stage‐Dependent Changes in the Diameter of the Lumen of the Rat Seminiferous Tubule
Authors: SHARPE RICHARD M.
Published: December 2015

Adult male rats were treated with a single dose of 650 mg/kg methoxy acetic acid to deplete the seminiferous tubules specifically of pachytene and later spermatocytes. The effect of this treatment and the subsequent maturation‐depletion of later germ cell types on the diameter of the seminiferous tubule and its lumen and the area of the seminiferous epithelium were studied in relation to the stages of the spermatogenic cycle. At 21 days after methoxy acetic acid treatment, the diameter of the tubule and the area of the epithelium were reduced below control values at all stages, consistent with the reduced number of early (stage VIII) or late (all other stages) spermatids. Unexpectedly, diameter of the lumen was also reduced at all stages other than VIII, and especially at stage VII. In controls, lumen diameter at stages VII and VIII was increased by ∼ 50% compared with earlier and later stages. In rats treated 21 days previously with methoxy acetic acid no change occurred at stage VII (lacking elongated spermatids) while a normal increase did occur at stage VIII (lacking round but not elongated spermatids). At earlier times after methoxy acetic acid treatment when stage VII tubules were depleted of pachytene spermatocytes alone (3 days) or together with early spermatids (7 days), the diameter of the lumen was not significantly different from the control value. These data suggest that lumen diameter may be regulated by elongated spermatids, especially at stages VII and VIII.
Mast Cells in the Testis, Epididymis and Accessory Glands of the Rat: Effects of Neonatal Steroid Treatment
Authors: GAYTAN F. , CARRERA G. , PINILLA L. , AGUILAR R. , BELLIDO C.
Published: December 2015

Mast cells in the testis of control adult rats were found almost exclusively around subcapsular blood vessels. Discrete mast cells were distributed throughout the stroma of the epididymis and sex accessory glands. In neonatally estrogen‐treated rats, a greater number of mast cells was present in the testicular interstitium, whereas no significant increase in the number of mast cells per square millimeter of stroma was found for the epididymis and sex accessory glands, despite stromal proliferation. On the other hand, androgen‐treated rats did not have increased mast cell numbers in any organ. These results indicate that the increase in mast cell numbers was estrogen‐dependent, specifically related to the testis and did not seem to be a consequence of the increase in the connective interstitial tissue.
Circadian Variation in Testosterone, Sex Hormone‐Binding Globulin, and Calculated Non‐Sex Hormone‐Binding Globulin Bound Testosterone in Healthy Young and Elderly Men
Authors: PLYMATE STEPHEN R. , TENOVER JOYCE S. , BREMNER WILLIAM J.
Published: December 2015

The circadian pattern in levels of serum total testosterone (T) in men becomes blunted with normal aging. However, because T not bound to sex hormone‐binding globulin (non‐SHBG‐T) is felt to be a better representative of biologically available T than is total T, the possibility of a 24‐h variation in non‐SHBG‐T in young men and the possibility that aging is associated with a blunting of that rhythm were investigated. Hourly blood samples were drawn on 10 normal young men (mean age 27.3 years) and 10 normal elderly men (mean age 70.7 years) over a 24‐h period and the serum was assayed for total T, sex hormone‐binding globulin (SHBG), and total protein; non‐SHBG‐T was calculated. SHBG was determined by radioimmunoassay as well as by a steroid‐binding assay. Young men had a significantly higher (p < 0.05) mean 24‐h level of non‐SHBG‐T (1.91 ± 0.62 nm/l) than did the elderly men (0.86 ± 0.01 nM/l). Also, each young man showed a significant circadian rhythm in non‐SHBG‐T, with a group mean daily variation of 1.42 ± 0.38 nM/l. In contrast, only 60% of the elderly men demonstrated a significant circadian rhythm in non‐SHBG‐T, and the group mean rhythm was blunted (maximum excursion 0.38 ± 0.07 nM/l) compared with that of the young men. SHBG and total protein levels demonstrated similar 24‐h variations in the two age groups. It was concluded that non‐SHBG‐T serum levels, similar to serum total T levels, demonstrate a circadian pattern in young men and this circadian rhythmicity becomes blunted with normal aging.