bluuebunny/crossref_metadata_2025 huggingface dataset:
Acute Effects and Long‐Term Sequelae of 1,3‐Dinitrobenzene on Male Reproduction in the Rat II. Quantitative and Qualitative Histopathology of the Testis
Authors: HESS REX A. , LINDER RALPH E. , STRADER LILLIAN F. , PERREAULT SALLY D.
Published: December 2015

This study determined the quantitative and qualitative histopathologic effects of a single oral dose of 1,3‐dinitrobenzene (48 mg/kg) on the rat testis from 1 to 175 days postexposure. The testis was damaged severely by hour 24, as evidenced by increased numbers of regressive seminiferous tubules that exhibited degenerating pachytene spermatocytes, chromatin margination in spermatids, giant cells, deformed spermatid heads, retained spermatids, and reduced numbers of meiotic figures. The major effects during the first 48 hours posttreatment were degeneration or exfoliation of pachytene spermatocytes and round spermatids and the retention of step 19 spermatids. These regressive effects continued until 24 days, after which the tubules either recovered or became atrophic. At the end of the study (175 days), three males were normal, one had regressed testicles, and three males had atrophic tubules (15 to 45%). Several cellular abnormalities were common throughout the period. In addition, the frequency of the stages of spermatogenesis was altered, an indication of a disturbance in the kinetics of spermatogenesis. 1,3‐Dinitrobenzene produced profound and specific lesions in the seminiferous tubules, and recovery was slow and incomplete. Atrophic tubules seemed to form if the normal cellular associations were not reestablished within 24 days, possibly due to the inability of Sertoli cells to reorganize the synchrony of germ cell development.
Radioautographic Localization of Atrial Natriuretic Factor Receptors in the Rat Testis
Authors: PELLETIER GEORGES
Published: December 2015

Recent data have suggested that atrial natriuretic factors (ANFs) can modulate Leydig cell functions. An in vitro radioautographic procedure on slide‐mounted frozen testicular sections was used to localize ANF receptors in rat testis. The radioligand was rat [125I]ANF‐28. It was demonstrated that ANF specific binding sites were present only in interstitial cells, and that the other testicular compartments were not significantly labeled. These results suggest that ANF could have some physiologic role in testicular functions.
The Pattern of LH Release in the Adult Ram Influences the Testicular Steroidogenic Response to Individual LH Pulses and is Regulated by Testosterone Negative Feedback
Authors: SANFORD LEE M. , PONZILIUS KARL‐HEINZ
Published: December 2015

Two experiments were conducted with adult intact rams (∼ 58 kg in body weight) in the nonbreeding season to investigate interrelationships between LH and testosterone secretion. In Experiment 1, LH pulse frequency was increased from ∼ two to six peaks per 8 hours (for 56 hours) by injecting (iv) 10 μg NIH‐LH‐S18 every 80 minutes. Induction of a breeding season peak frequency produced a progressive 3‐fold increase (P < 0.01) in mean serum testosterone levels to values during the last 8 hours of treatment (12.6 ± 1.2 ng/ml) that were 50% of those in the fall. In response to LH pulsing, testosterone peak amplitude increased (P < 0.05) from 3.5 ± 0.8 ng/ml to 6.7 ± 0.7 ng/ml. In Experiment 2, the mean testosterone level was increased to breeding season values (for 96 hours) by injecting (im) 5 mg testosterone every 4 hours. Mean LH levels and LH peak frequency were decreased (∼ 70%, P < 0.01) following 36 hours of treatment, and the LH response to exogenous GnRH was decreased (∼ 45%, P < 0.01) by the final 4 hours Results indicate that for rams in the nonbreeding season, the testicular steroidogenic response to individual LH pulses is enhanced when pulse frequency is increased. When blood testosterone is elevated to breeding season levels, LH pulse frequency is severely impaired, while pituitary responsiveness to GnRH is diminished, as in the fall.
Two‐Dimensional Gel Electrophoretic Analysis of Rat Sperm Membrane Interaction with Cauda Epididymal Fluid
Authors: HALL JOSEPH C. , KILLIAN GARY J.
Published: December 2015

Two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE) was used to analyze the polypeptide composition of rat cauda epididymal fluid, blood serum and membrane‐enriched fractions of caput, corpus, and cauda epididymal spermatozoa. Several polypeptides were found in both cauda fluid and blood serum, and in both cauda fluid and epididymal spermatozoa. Prominent cauda epididymal fluid polypeptides that were associated with caput, corpus, and cauda sperm membranes were 32 and 33 kDa. Passage of spermatozoa from the caput to the cauda epididymidis was characterized by the loss of three glycopolypeptides of 32, 30 and 29 kDa, and by the addition of a 37‐kDa glycopolypeptide. Incubation of intact caput, corpus and cauda spermatozoa with cauda epididymal fluid revealed major changes in the polypeptide maps of the incubation fluid and the membrane‐enriched fractions of caput and corpus, but not cauda spermatozoa. The incubation of cauda fluid with caput and corpus sperm cells was characterized by a loss of several polypeptides and the addition of a 24‐kDa glycopolypeptide. The most striking change in spermatozoa incubated with cauda epididymal fluid was the addition of two glycopolypeptides of 32 and 33 kDa to the polypeptide maps of caput sperm cells. These data demonstrate that rat spermatozoa undergo surface modifications during epididymal maturation and that these modifications can be influenced by epididymal fluid.
Leydig Cells Do Not Have Fc Receptors
Authors: HUTSON JAMES C.
Published: December 2015

It has been reported that Leydig cells have Fc receptors, which traditionally have been considered markers specific for macrophages and polymorphonuclear leukocytes. The purpose of this investigation was to study further this phenomenon and also to determine if Leydig cells and macrophages could be separated from each other either by density gradient centrifugation using Percoll or by differential detachment with trypsin treatment of cultures of crude interstitial preparations. Interstitial cells were obtained by collagenase digestion of rat testis and established in culture. These cultures were reacted for 3β‐steroid dehydrogenase and Fc receptor and viewed with phase contrast microscopy. No individual cells were positive for both steroid dehydrogenase activity and Fc receptors. The order in which the cells were stained for these two markers did not influence the results. Trypsin treatment of these crude interstitial cultures removed over 90% of the Leydig cells and approximately 20% of the macrophages. Macrophages were located in the same portion of Percoll gradients as the less dense (Population I) Leydig cells, while Leydig cells found in the dense area of the gradient (Population II) were not contaminated with macrophages. These studies indicate that Leydig cells do not have Fc receptors and that a subpopulation of Leydig cells can be isolated free of macrophages using density gradient centrifugation.
Testosterone Dose‐Dependency of Sexual and Nonsexual Behaviors in The Gonadotropin‐Releasing Hormone Antagonist‐Treated Male Rat
Authors: FIELDER THOMAS J. , PEACOCK NADINE R. , GIVERN ROBERT F. MC , SWERDLOFF RONALD S. , BHASIN SHALENDER
Published: December 2015

The testosterone dose‐dependency of several mating and nonmating behaviors was examined in the male rat, chemically castrated with a GnRH antagonist analog. Graded doses of testosterone enanthate (TE) were given to male rats to reinstate behaviors abolished by GnRH antagonist treatment. GnRH antagonist treatment alone markedly lowered serum LH, FSH and T concentrations and ventral prostate and testis weights. Open field behaviors were not significantly affected by GnRH antagonist treatment or castration. Scent‐marking behavior was markedly suppressed by both castration and GnRH antagonist and restored by the lowest dose of TE (0.05 mg). All measures of male sexual behavior were impaired by GnRH antagonist treatment and castration and restored by the lowest dose of TE (0.05 mg). The doses of TE required to restore normal ventral prostate weights and testis weights were higher than those required to maintain scent marking and mating behaviors. No direct behavioral effects of the GnRH antagonist, other than those that can be explained by GnRH antagonist‐induced suppression of testosterone were observed. The finding that sexual and nonsexual behaviors in the male rat have different testosterone requirements from those maintaining spermatogenesis and fertility may have significant implications for contraception.
Incorporation of Gossypol and Formation of Its Protein Conjugates in Mouse Transformed Sertoli (TM4) Cells
Authors: TANPHAICHITR NONGNUJ , FITZGERALD LISA M.
Published: December 2015

Mitochondria of both mouse transformed Sertoli (TM4) cells and primary‐cultured rat and mouse Sertoli cells were shown to be preferentially affected by gossypol (Tanphaichitr et al, 1984). To investigate whether this selective effect was due to a greater ability of TM4 cells to accumulate gossypol, TM4 and other somatic cell lines, including dog (MDCK) and kangaroo (PtK2) kidney epithelial cells, rat embryo fibroblasts (Rat‐1) and mouse BALB/c 3T3 fibroblasts, were incubated with [14C]gossypol and the incorporated specific activity of the drug was assessed. The results indicate that TM4 cells accumulated [14C]gossypol at the highest level. Incorporated [14C]gossypol appeared to bind to TM4 cell macromolecules and remained in the dialysis tubing after extensive dialysis. Characterization of these gossypol‐conjugated proteins by SDS‐polyacrylamide gel electrophoresis followed by fluorography revealed that these proteins had apparent Mr's of 92,500, 70,000, 63,200, 60,000, 58,100, 54,000, 52,000, 50,000, 47,500, 40,000, 37,000, 35,000, 30,000, 20,000, and 14,500 daltons. Conjugation of these proteins with gossypol may result in macromolecular dysfunction and abnormal structure.
Spermatogenetic Expression of RNA‐Binding Motif Protein 7, a Protein That Interacts With Splicing Factors
Authors: Guo Taylor B. , Boros Laszlo G. , Chan Kam C. , Hikim Amiya P. Sinha , Hudson Alan P. , Swerdloff Ronald S. , Mitchell Aaron P. , Salameh Wael A.
Published: December 2015

ABSTRACT:We have previously shown that a ubiquitously expressed RNA splicing factor, RNA‐binding motif 7 (RBM7), cloned from a testis complementary DNA library, enhances messenger RNA (mRNA) splicing in vitro and is expressed in a cell‐restricted fashion. Herein, we detail its mRNA and protein expression in the rodent testis. RNA in situ hybridization shows that Rbm7 expression in rat germ cells closely parallels the entry and progression of meiosis. The expression commences in type B spermatogonia, it rises during the preleptotene stage, peaks in leptotene spermatocytes, and declines afterward, but increases again in stage‐associated pachytene spermatocytes. An affinity‐purified polyclonal antibody raised against a peptide corresponding to amino acids 202–224 of the mouse RBM7 recognized the predicted 35 kd protein both in testicular lysates and in in vitro translation reactions. Consistent with the in situ hybridization results, RBM7 immunoreactivity was also detected in type B spermatogonia, spanned the entire period of spermatocyte development, and extended to round and early elongated spermatids. Moreover, RBM7 appeared nuclear up to the mid pachytene stage and became cytoplasmic thereafter. Consistent with its role in RNA splicing, yeast 2‐hybrid and glutathione S‐transferase pull‐down assays show that RBM7 interacts with splicing factor 3b subunit 2 (SAP145), and with the splicing regulator, SRp20. These interactions and the nuclear localization of RBM7 provide insights into its function in pre‐mRNA processing in developing spermatocytes during entry into meiosis and progression through the meiotic prophase.
Molecular Analysis of the Androgen Receptor in Ten Prostate Cancer Specimens Obtained Before and After Androgen Ablation
Authors: Lamb Dolores J. , Puxeddu Efisio , Malik Nusrat , Stenoien David L. , Nigam Rajni , Saleh George Y. , Mancini Michael , Weigel Nancy L. , Marcelli Marco
Published: December 2015

ABSTRACT: Hormonal or androgen‐ablation (AA) therapy is the predominant form of systemic treatment for metastatic prostate cancer. Although an initial response to AA is observed in 70%–80% of patients with advanced disease, most tumors eventually progress to androgen‐independent growth, and only a minority of affected individuals are alive 5 years following initiation of treatment. Because AA induces a dramatic change in the hormonal milieu of the patient and because these tumors maintain the ability to proliferate, it is possible that this treatment selects a population of cells with mutated androgen receptors (ARs) that sustain growth despite the absence of circulating androgen. To test this hypothesis we investigated the molecular structure of the AR in 10 prostate cancer specimens obtained before and after AA. Tumors (coded A through L) were microdissected to uniquely enrich genomic DNA from cancer cells. Exons 1–8 of the AR were screened by polymerase chain reaction, single‐stranded conformational polymorphism, and sequence analysis. A mutation consisting of an expansion of the polyglutamine (poly‐Q) repeat from 20 (found in 100% of the sequences of specimens obtained before AA) to 26 (found in 70% of the sequences of specimens obtained after AA) was detected in patient F. The 26 glutamine (Q26) AR readily translocated to the nucleus upon addition of androgen, and did not show significant differences in its ability to bind 3[H]‐dihydrotestosterone compared to its wild‐type counterpart. Nevertheless, analysis of transcriptional activity showed that the Q66 AR was transcriptionally 30%–50% less active than the wild‐type molecule. Because clones of AR with an expanded poly‐Q tract were detected only in the specimen from patient F obtained after AA, we conclude that in specific circumstances, AA treatments can select variant forms of the AR in the prostate of patients affected by prostate cancer. Further experiments are needed to conclusively determine whether the Q26 clone was responsible for sustaining survival of prostate cancer cells in the androgen‐depleted milieu of this patient.
Effect of Azorellanone, a Diterpene FromAzorella yaretaHauman, on Human Sperm Physiology
Authors: Morales Patricio , Kong Milene , Pizarro Eduardo , Pasten Consuelo , Morales Glauco , Borquez Jorge , Loyola Luis A.
Published: December 2015

ABSTRACT:Previous studies have shown that cyclic terpenes extracted from plants decrease sperm motility and concentration in rats. In this work, we studied the effect 13‐α‐hydroxy‐7‐oxoazorellano (azorellanone), a cyclic diterpene extracted from Azorella yareta Hauman, on in vitro human sperm physiology. Sperm aliquots, capacitated for 4.5 or 20 hours, were incubated for 15 minutes with different concentrations of azorellanone. Then, the effects of azorellanone on sperm motility, viability, binding to the human zona pellucida, progesterone‐induced acrosome reactions and increase in intracellular Ca2+concentration, and trypsin and chymotrypsin‐like protease activities were evaluated. Sperm motility was evaluated according to World Health Organization (WHO) guidelines; sperm viability with the supravital dye Hoescht 33258; sperm—zona binding with the hemizona assay; progesterone‐induced acrosome reaction with fluorescent lectin; intracellular Ca2+level with fura 2; and protease activity with the synthetic substrates N‐t‐Boc‐Gln‐Ala‐Arg‐Amido‐4‐methylcoumaryn and Succinyl‐Leu‐Leu‐Val‐Tyr‐Amido‐4‐methylcoumaryn. The results obtained indicate that azorellanone inhibited sperm motility in a concentration‐dependent manner at 0.15, 1.5, and 3 mM, while sperm viability was only inhibited at 3 mM. Treatment with azorellanone significantly inhibited sperm—zona binding, progesterone‐induced acrosome reactions, and intracellular Ca2+concentration. Treatment of free‐swimming sperm with azorellanone did not affect protease activity; however, the incubation of sperm extracts with azorellanone significantly inhibited both trypsin‐like and chymotrypsin‐like protease activities. In conclusion, azorellanone has a significant effect on the different parameters that characterize human sperm physiology.
Status of Spermatogenesis and Sperm Parameters in Langur Monkeys Following Long‐term Vas Occlusion With Styrene Maleic Anhydride
Authors: Mishra Pradyumna K. , Manivannan B. , Pathak Neelam , Sriram S. , Bhande Satish S. , Panneerdoss S. , Lohiya Nirmal K.
Published: December 2015

ABSTRACT: Vas occlusion by styrene maleic anhydride (SMA), trade name RISUG (one of the promising male contraceptive procedures currently in phase III clinical trials), at 60 mg/vas deferens dissolved in 120 μL dimethyl sulphoxide (DMSO) at up to a 540‐day study period caused severe oligospermia in the first 2 to 3 ejaculations and uniform azoospermia in the subsequent ejaculations without toxicity in langur monkeys. The ejaculated spermatozoa were necroasthenoteratozoospermic, suggesting instant sterility. Routine hematology and clinical chemistry parameters and the serum testosterone and sperm antibody titers remained unchanged from their pretreatment values until 540 days vas occlusion. Histology of testes revealed continued spermatogenesis throughout the study period. The stages of spermatogenesis appeared normal until 300 days of vas occlusion. At 360 days of vas occlusion, germ cells appeared in the lumen. Degeneration of seminiferous epithelium was evident in some of the tubules. Following 420 days of vas occlusion, the central portion of the testis showed regressed seminiferous tubules depicting various shapes and devoid of germ cells, which continued until 540 days of vas occlusion. Ultrastructure of the testes after 540 days of vas occlusion revealed vacuolization in the cytoplasm of Sertoli cells and degenerative features in the membranes of the spermatocytes and spermatids in the affected seminiferous tubules. The sub‐cellular features of the normal tubules were similar to those of controls. The results suggest focal degeneration of seminiferous epithelium in the central portion of the testis following long‐term vas occlusion with SMA.
Role of Tissue Inhibitor of Metalloproteases‐1 in Junction Dynamics in the Testis
Authors: Mruk Dolores D. , Siu Michelle K.Y. , Conway Anne M. , Lee Nikki P.Y. , Lau Ann S.N. , Cheng C. Yan
Published: December 2015

ABSTRACT:Using multiple high‐performance liquid chromatography steps, we have identified and purified a polypeptide to apparent homogeneity from primary Sertoli cell conditioned culture medium that consisted of 2 molecular variants of 31 and 29 kDa when electrophoresed on a sodium dodecyl sulfate—polyacrylamide gel run under reducing conditions. Partial N‐terminal amino acid sequence analysis of these 2 proteins revealed a sequence of NH2‐IKMAKMLKGFDAVGNATG, which is homologous to tissue inhibitor of metalloproteases‐1 (TIMP‐1). Studies by semiquantitative reverse transcription‐polymerase chain reaction using a primer pair specific to rat TIMP‐1 demonstrated that both Sertoli and germ cells express TIMP‐1. During maturation, the steady‐state TIMP‐1 mRNA level in the testis increased significantly from 40 to 60 days of age, which suggests its role in the restructuring of the epithelium during spermiation. This increase in testicular TIMP‐1 expression was apparently not due to the increase in germ cell number, because TIMP‐1 expression decreased approximately fivefold in germ cells isolated from testes of aging rats. Using Sertoli cells cultured at low (0.05 × 106cells/cm2) and high (0.5 × 106cells/cm2) densities, it was found that TIMP‐1 expression increased transiently but significantly during junction assembly. A similar induction of TIMP‐1 mRNA was also detected in Sertoli—germ cell cocultures during germ cell adhesion onto Sertoli cells. More important, the inclusion of either α2‐macroglobulin (a protease inhibitor produced by Sertoli cells) or aprotinin (a serine protease inhibitor) into an in vitro germ cell adhesion assay facilitated the attachment of fluorescently labeled germ cells onto the Sertoli cell epithelium when compared to control, which suggests that the assembly of adherens junctions may involve protease inhibitors.
Reduced Human Germ Cell‐Less (HGCL) Expression in Azoospermic Men With Severe Germinal Cell Impairment
Authors: Kleiman Sandra E. , Yogev Leah , Gal‐Yam Einav Nili , Hauser Ron , Gamzu Ronni , Botchan Amnon , Paz Gedalia , Yavetz Haim , Maymon Batia Bar‐Shira , Schreiber Letizia , Barzilai Shlomit , Amariglio Ninette , Rechavi Gideon , Simon Amos J.
Published: December 2015

ABSTRACT: Germ cell‐less (GCL) protein is a nuclear envelope protein highly conserved between the mammalian and Drosophila orthologues. In Drosophila, maternal GCL protein is required to establish the germ lineage during embryonic development. In mammals, it is suggested that the GCL function is mainly in spermatogenesis and that it might be related to the ability of mouse GCL to repress transcription. Using reverse transcriptase‐polymerase chain reaction analyses, we investigated the role of human GCL (HGCL) in spermatogenesis by studying its expression in the testicular tissue of 67 azoospermic men with normal karyotype and no Y‐chromosome microdeletion. Their testicular biopsy specimens underwent meticulous histological and cytological analysis as well as molecular analysis with various markers of spermatogenesis (RBM1, DAZ, and CDY1). The rate of X‐Y and 18 chromosome bivalent formation during meiosis was additionally assessed in 22 of these biopsy specimens and correlated to HGCL expression. Expression of HGCL was affected in parallel with the severity of testicular impairment found. Defective sperm motility was associated with the absence of HGCL. Nevertheless, the absence of HGCL expression did not influence the normal process of chromosome bivalent formation in meiosis. Our results suggest that HGCL is not essential for the chromosomal events of meiosis but might be involved in later aspects of spermatogenesis.
Oral Testosterone‐Triglyceride Conjugate in Rabbits: Single‐Dose Pharmacokinetics and Comparison With Oral Testosterone Undecanoate
Authors: Amory J. K. , Scriba G. K. E. , Amory D. W. , Bremner W. J.
Published: December 2015

ABSTRACT: Development of a safe and effective oral form of testosterone has been inhibited by the rapid hepatic metabolism of nonalkylated androgens. Since triglycerides are absorbed via lymphatics and bypass the liver, we hypothesized that a testosterone‐triglyceride conjugate (TTC) might allow for safe and effective oral testosterone therapy. Therefore, we studied the single‐dose pharmacokinetics of oral administration of TTC in rabbits. Female New Zealand rabbits were administered 2, 4, or 8 mg/kg of TTC in sesame oil by gastric lavage. Testosterone undecanoate (TU) by gastric lavage was used as a positive control. Blood was sampled from a catheter in the auricular artery at 0, 15, 30, 60, 90, 120, 180, 240, 360, 480, and 600 minutes after drug administration. Samples were assayed for testosterone by a fluoroimmunoassay. Mean serum testosterone, area under the curve (AUC), and terminal half‐life were calculated. Oral TTC administration resulted in rapid and marked increases in serum testosterone. Oral TTC resulted in higher maximum serum testosterone concentrations than oral TU at 8 mg/kg (TTC: 28.6 ± 7.9 nmol/L vs TU: 11.9 ± 2.1 nmol/L; P < .001) and 4 mg/kg (TTC: 11.5 ± 4.2 nmol/L vs TU: 3.6 ± 1.0 nmol/L; P < .001). In addition, the AUC was 1.8 to 2.6 times greater for TTC than TU at both doses (P < .05). The terminal half‐life for both TU and TTC was between 3 and 5 hours and was not significantly different. We conclude that oral TTC is rapidly absorbed from the rabbit intestine and results in elevated concentrations of serum testosterone. The absorption of TTC appears to be superior to that of TU; however, the in vivo persistence of the 2 compounds is similar. TTC may offer an alternative to the use of TU for oral testosterone therapy. Further testing of this compound is warranted.
Expression and Localization of Total Akt1 and Phosphorylated Akt1 in the Rat Seminiferous Epithelium
Authors: Hixon Mary L. , Boekelheide Kim
Published: December 2015

ABSTRACT: Akt1 is a survival factor that is believed to play a role in the transcriptional modulation of a subset of genes associated with cell growth, proliferation, and apoptosis. We have explored in detail the expression of total Akt1 and phosphorylated Akt1 in the developing and adult rat testis. Throughout testis postnatal development, the expression of total Akt1 protein exhibited a mainly cytoplasmic localization within both the germ cells and the supporting Sertoli cells. In contrast, phosphorylated Akt1 staining demonstrated a mainly nuclear localization within germ cells. In the developmental sequence of germ cells, phosphorylated Akt1 stained the nuclei of spermatogonia, spermatocytes, and round spermatids. During spermiogenesis, phosphorylated Akt1 staining decreased in the nucleus and became localized to a bright spot at the base of the nucleus in elongate spermatids. Of interest, total Akt1 was found to localize to the perinuclear region of germ cells and the supranuclear region of Sertoli cells, depending on fixation. Further analysis demonstrated this staining to be associated with the Golgi complex in both germ and Sertoli cells.
Abundance of Repetitive Sequence Elements in the Mouse Testis‐Specific Lactate Dehydrogenase‐C Gene1
Authors: Olsson Per G. , Tsujioka Hiroshi , Narisawa Sonoko , Goldberg Erwin , Millán José Luis
Published: December 2015

ABSTRACT: We have cloned and sequenced the entire mouse ldhc gene and mapped it physically in relation to the somatic ldha gene. The 2 genes were found to be oriented in head‐to‐tail fashion with about a 6‐kilobase (kb) distance between the 3′ end of ldha and the 5′ end of ldhc. The ldhc gene is composed of 43% repetitive elements compared to only 16% in the ldha gene. Despite the close physical distance of mouse ldha and ldhc, the 2 genes have a very different content of repetitive elements, and this most likely reflects different levels of selective pressure.
Vasectomy Influences Expression of HE1 but not HE2 and HE5 Genes in Human Epididymis
Authors: Légaré Christine , Verville Nancy , Sullivan Robert
Published: December 2015

ABSTRACT: Epididymal roles include protection and transport, maturation, and storage of the sperm cells. It is known that these functions are altered under vasectomy, but the consequences of excurrent duct obstruction on the pattern of gene expression along the human epididymis are poorly documented. In order to understand how excurrent duct occlusion affects different epididymal regions, the expression pattern of genes known to be expressed in specific epididymal segments was investigated along the epididymides of vasectomized men. Selected human epididymal complementary DNAs (cDNAs) identified by differential library screening were studied because of their unique messenger RNA (mRNA) distribution along the different epididymal segments. In situ hybridization as well as immunohistologic studies were undertaken to investigate the effect of vasectomy on a gene expressed all along the epididymis (HE1) or more selectively in the proximal (HE2) or distal (HE5) segment. The HE1 transcript was affected by the obstruction of the epididymis with little or no mRNA detectable along the epididymis. The HE1‐related antigen was shown by immunohistochemical methods to be reduced within the epithelium of the epididymis of vasectomized men. By contrast, HE5 mRNA and protein, expressed in epithelial cells of the distal epididymis, were not affected by the obstruction of the vas deferens. Similarly, HE2 transcriptional and translational products normally expressed in the caput epididymidis were not affected by vasectomy. These results show that excurrent duct obstruction differentially affects the expression pattern of some specific transcripts and their encoded proteins, probably impairing their fundamental roles in the physiology of the epididymis.
Functional Analysis of the Cooled Rat Testis
Authors: Zhang Z. , Short R.V. , Meehan T. , de Kretser D.M. , Renfree M.B. , Loveland K.L.
Published: December 2015

ABSTRACT:Direct cooling of the testis results in the depletion of most germ cells in vivo. Germ cell‐depleted testes are now commonly used to investigate spermatogenic regeneration and can serve as recipients for germ cell transplantation. The present study explored the effects of cooling rat testes on the depletion of endogenous germ cells, spermatogenic regeneration, and Sertoli cell function. Adult rat testes were cooled with iced Ringer's solution for 60 minutes, which results in the initiation of apoptotic germ cell loss within 8 hours. Pachytene spermatocytes at stages XII‐I were the cells most sensitive to cooling. In 46%‐67% of seminiferous tubule cross‐sections, only Sertoli cells remained in the cooled testes 3–10 weeks after treatment. Germ cell loss was accompanied by a significant decrease in circulating inhibin B and an increase in follicle‐stimulating hormone concentrations, which indicated a change in Sertoli cell function. Quantitative analysis of mRNA expression associated with apoptotic signals showed no significant uniform changes among the cooled testes, although some individuals had a distinct up‐regulation of FAS mRNA at 24 hours. Attempts to use the cooled testes as recipient testes for mouse‐to‐rat germ cell transplantation were undertaken, but none of the mouse germ cells transplanted into the testes 15–34 days after cooling appeared to have undergone spermatogenesis 64–92 days after transplantation. These data suggest that modifications to Sertoli cell function resulting from testicular cooling create an environment that is unable to support spermatogenesis by donor germ cells.
Selection of Individual Testicular Tubules From Biopsied Testicular Tissue With a Stereomicroscope Improves Sperm Retrieval Rate1
Authors: Kamal Ahmed , Fahmy Ibrahim , Mansour Ragaa T. , Abou‐Setta Ahmed M. , Serour Gamal I. , Aboulghar Mohamed A.
Published: December 2015

ABSTRACT: The present study describes a new modification for testicular sperm extraction (TESE) with an intraoperative surgical loop, coupled with stereomicroscopic dissection in the laboratory, to identify sperm‐containing tubules. The study included 116 consecutive patients with nonobstructive azoospermia (NOA) undergoing TESE and intracytoplasmic sperm injection. After dissection of testicular tissue under the stereomicroscope, patients were separated into 2 groups according to tubular diameter. In the first group (n = 72), all tubules were of the same diameter. In the second group (n = 44), tubules with variable diameters could be identified. In such cases, 1–2 of the most distended and opaque seminiferous tubules were selected and minced alone, then examined for the presence of spermatozoa. The rest of the testicular tissue suspension was minced and examined separately. In 11 (25%) cases, retrieved spermatozoa were found in the isolated distended tubules only. In 21 (47.7%) cases, spermatozoa were found in both the isolated distended tubules and the rest of the specimen. In 2 cases, spermatozoa were found only in the whole sample, not in the isolated tubules. In the remaining 10 cases, spermatozoa were not found in either the whole sample or the isolated tubules. The sperm recovery rate in the isolated tubules was significantly higher than that of the rest of the specimen (72.7% vs 52%, χ2 = 3.93, P < .05), and larger numbers of spermatozoa could be easily retrieved in a shorter period of time. In conclusion, the selection and isolation of the most dilated and opaque seminiferous tubules by using the surgical loop, coupled with laboratory stereoscopic dissection, improves sperm retrieval for men with NOA. It is possible that surgical‐loops TESE coupled with stereomicroscope may offer superior sperm retrieval when compared with conventional TESE and may also offer reduced operative time when compared with microdissection TESE.
Trainee Page: Careers in Andrology
Authors: Bomgardner Daniela , Comizzoli Pierre
Published: December 2015

ABSTRACT: As you are reading this issue, you should be making plans to attend the American Society of Andrology's 29th Annual Meeting to be held in Baltimore, Md, from April 17 to April 20, 2004. This is a great event to learn as well as to mix and mingle with fellow trainees and distinguished professionals in the field. For those unsure of their future career path, it is a fantastic opportunity to socialize with several members who have taken a nontraditional approach to a conventional degree. Below, we have singled out 2 individuals who were generous enough to tell their story of alternative career paths, but there are certainly more. Take the time to search these people out as well as any one of our Society members. They are a tremendous resource to answer your relevant topics such as grant writing, searching for a post‐doc position or job, taking alternative career paths, or succeeding in the clinic or laboratory.
Trainee Page
Authors: Palladino Michael A. , Vogel Donna L.
Published: December 2015

ABSTRACT: To the graduating class of 2004, we extend our congratulations and wish you much luck in the future. Remember, the American Society of Andrology is a useful tool to be utilized in your job hunt. To aid in your quest, we thought we would offer you some insight in how to go about that search. Take a look at Dr Michael Palladino's piece on faculty positions. For all those still plugging away, this is a good piece for you to hold onto until your time. To everyone, enjoy the summer. Hit the beach with a good book or the Journal article by Dr Donna Vogel who manages to debunk that great mystery we know as grants. Whatever you do have a great summer!
Evidence for the Presence of Angiogenin in Human Testis
Authors: Koga Kaori , Osuga Yutaka , Yano Tetsu , Ikezuki Yumiko , Yoshino Osamu , Hirota Yasushi , Hirata Tetsuya , Horie Shigeo , Ayabe Takuya , Tsutsumi Osamu , Taketani Yuji
Published: December 2015

ABSTRACT: We have reported the expression and possible roles of angiogenin, a potent angiogenic factor, in human female reproductive organs. In this study, we investigated the expression of angiogenin in the human testis, a male reproductive organ. Western blot analysis showed the presence of angiogenin in the human testis, with a single band of the same size as recombinant human angiogenin. Immunohistochemical study and in situ hybridization showed that the angiogenin protein and messenger RNA (mRNA) localized in peritubular myoid cells (PTMCs) and vascular endothelial and smooth muscle cells. PTMCs are known to play various roles in the testes concerned with spermatogenesis, transport of spermatozoa, structural support to the seminiferous tubules, and mediation of Sertoli cell function. The specific localization of angiogenin in PTMCs suggests that angiogenin plays physiologic roles in the human testis.
Effects of Centrifugation Before Freezing on Boar Sperm Cryosurvival
Authors: Carvajal Gema , Cuello Cristina , Ruiz Maria , Vázquez Juan María , Martínez Emilio Arsenio , Roca Jordi
Published: December 2015

ABSTRACT: Current protocols for boar sperm cryopreservation require the centrifugation of semen in order to separate sperm cells from the seminal plasma. This study evaluated the influence of different centrifugation regimes on both sperm recovery and yield (percentage of viable sperm with an intact acrosome relative to the initial sperm population) after centrifugation (experiment 1) as well as the influence of different centrifugation regimes on boar sperm cryosurvival (experiment 2). In both experiments, sperm‐rich fractions from 3 boars were diluted, pooled, and cooled to 17°C before centrifugation. In experiment 1, the g‐forces tested were 400, 800, 1600, and 2400 × g for 3 or 5 minutes, using the standard regime (800 × g for 10 minutes) as a reference. Sperm recovery (Bürker Chamber) and yield (triple fluorescent stain of PI/R123/FITC‐PNA [DNA‐specific fluorochrome propidium iodide/mitochondria‐specific fluorochrome rhodamine‐123/acrosome‐specific fluorochrome fluorescein isothiocyanate‐labeled peanut (Arachis hypogaea) agglutinin]) were calculated. The highest recovery and yield (P < .05) values were achieved using 2400 × g for 5 or 3 minutes and 1600 × g for 5 minutes, which showed no differences (P > .05) from the reference in terms of sperm yield. In experiment 2, cooled semen was centrifuged using 3 different regimes: C1 (2400 × g for 3 minutes), C2 (1600 × g for 5 minutes), and C3 (800 × g for 10 minutes). Pellets were diluted in lactose‐egg yolk (LEY)‐glycerol‐Equex STM (1 × 109 cells/mL) and frozen in 0.5‐mL straws. After thawing, sperm quality was assessed after 30 and 150 minutes of incubation (37°C). Centrifugation regimes C1 and C2 showed significantly (P < .05) higher postthaw sperm motility (assessed with a computer‐assisted semen analysis system), viability (evaluated as for experiment 1), and percentage of uncapacitated sperm (assessed with a chlortetracycline assay) than did C3. In addition, C1 had the highest (P < .05) oocyte penetrating ability (assessed with the homologous in vitro penetration test performed with immature oocytes). Malondialdehyde production, assessed with the thiobarbituric acid reactive species test, was unaffected (P > .05) by the centrifugation regime used. We conclude that high g‐force (2400 × g) and short centrifugation time (3 minutes) do not affect sperm recovery and yield and that, moreover, they have a positive effect on the cryosurvival of boar sperm. Therefore, we recommend the use of short‐term centrifugation with a relatively high g‐force (2400 × g for 3 minutes) in boar sperm cryopreservation protocol.
Androgen Regulation of JM‐27 Is Associated With the Diseased Prostate
Authors: Shah Uzma S. , Arlotti Julie , Dhir Rajiv , Lu Sun , Pirozzi Gregorio , Prakash Kulkarni , Getzenberg Robert H.
Published: December 2015

ABSTRACT: Despite intense research efforts, the etiology of prostatic hyperplasia associated with both benign prostatic hyperplasia (BPH) and prostate cancer remains poorly understood. Our previous studies using array technology identified JM‐27 as a transcript that is dramatically up‐regulated in the prostates of patients with symptomatic BPH and in normal, adjacent prostatic regions of patients with prostate cancer. In the present study, using an extended sample set, we show a correlation between the messenger RNA and protein expression of JM‐27. To investigate the possible functions of this gene, its expression in the rat prostate was examined by immunoblot analysis using a polyclonal antibody specific to human JM‐27. This antibody reacts with 2 rat polypeptides of 17 kd and 27 kd in size. Whereas the 27‐kd form of the JM‐27 protein found in human prostate is selectively expressed in the dorsolateral lobes of the rat prostate, the 17‐kd form is expressed only in the ventral lobe. Expression of both forms of this protein appears to be androgen‐regulated. There is a time‐dependent decrease in expression of the protein products in the ventral and dorsolateral lobes of the rat prostate after castration. Administration of exogenous testosterone in castrated animals maintains protein expression in both lobes. Androgens are believed to play a central role in prostate growth and development, and therefore, it is tempting to speculate that JM‐27, an androgen‐regulated gene, may be involved in prostatic growth regulation. Further studies are underway to evaluate such a function for JM‐27 in prostatic diseases.
Glu298Asp Endothelial Nitric Oxide Synthase Polymorphism Is a Risk Factor for Erectile Dysfunction in the Mexican Mestizo Population
Authors: Rosas‐Vargas Haydee , Coral‐Vazquez Ramon M. , Tapia Rosario , Borja Jose L. , Salas Ricardo A. , Salamanca Fabio
Published: December 2015

ABSTRACT: Penile erection depends on the balanced action between antagonist vasoactive molecules such as nitric oxide (NO) and angiotensin. Endothelial nitric oxide synthase (eNOS) and angiotensin‐converting enzyme (ACE) polymorphisms have been associated with endothelial dysfunction, which is described as a cause of erectile dysfunction (ED). Endothelial NOS and ACE are both regulators of vascular and corporal smooth muscle tone, which are connected by interaction between the NO‐cyclic guanosine monophosphate pathway and the renin‐angiotensin system. We analyzed the frequencies of 894 G/T (Glu298Asp) eNOS and ACE I/D polymorphisms in Mexican patients with ED (n = 53) and in an age‐matched control group (n = 62). The populations analyzed were in Hardy Weinberg equilibrium. We found significant differences in allelic (χ2 = 4.42; P = .03) and genotypic frequencies (χ2 = 3.96; P = .04) between patients and controls for the 894 G/T eNOS polymorphism. Presence of the 894T allele in carriers increased the risk of ED (odds ratio [TT + GT versus GG] = 2.37; 95% confidence interval, 1.08 to 5.21; P = .02). Multiple logistic regression analysis showed that the Glu298Asp polymorphism was an independent factor for ED, as was diabetes mellitus, hypertension, cardiac disease, and cigarette smoking. No association was found between ACE I/D polymorphism and ED in the population studied. Therefore, our results suggest that Glu298Asp eNOS polymorphism plays a role as a genetic susceptibility factor for ED.
Sperm Membrane Dynamics Assessed by Changes in Lectin Fluorescence Before and After Capacitation
Authors: Baker Sarah S. , Thomas Monzy , Thaler Catherine D.
Published: December 2015

ABSTRACT: Sperm capacitation is correlated with acquisition of fertilizing ability, and the molecular events underlying this process are only beginning to be understood. A number of membrane changes associated with capacitation have been documented. In this study we used lectin probes to identify changes in glycoprotein localization as a result of capacitation of mouse sperm. Eight lectins (LEA, PSA, PNA, AAA, UEA‐1, WGA, STA, and TPA) stained regions of the mouse sperm head, tail, or both. No changes in tail staining patterns were detected when sperm were incubated under capacitating conditions. In contrast, 7 of 8 lectins tested showed clear shifts in staining patterns in the sperm head as a result of incubation under capacitating conditions. When staining patterns were quantified, a distinct heterogeneity within the sperm population was observed. Each lectin displayed 3 distinct staining patterns in both uncapacitated and capacitated sperm samples. The least common pattern represented the acrosome‐reacted (AR) pattern, as independently assessed by lectin staining of ionophoretreated sperm that were >95% AR as judged by Coomassie staining. However, a reciprocal shift in the two predominant staining patterns was correlated with capacitation and suggests that changes in distribution of cell surface proteins during capacitation constitute part of the molecular changes which result in changes in sperm function acquired during this process.
The Effect of Electrocoagulation on the Sinusoids in the Human Penis
Authors: Hsu Geng‐Long , Hsieh Cheng‐Hsing , Wen Hsien‐Sheng , Hsu Wen‐Long , Chen Yi‐Chang , Chen Robert M. , Chen Shyh‐Chyan , Hsieh Ju‐Ton
Published: December 2015

ABSTRACT: We give an overview of patients who have undergone removal of the deep dorsal vein for venous grafting in treating Peyronie disease with or without a Bovie effect. From June 1998 to May 2002, 23 men received grafting of the deep dorsal vein for morphologic correction. Among them, 7 men underwent electrocoagulation treatment of bleeders per surgeons' customary practice during the entire procedure and were categorized as the electrocoagulation group. Sixteen patients received simple ligation of bleeding stumps, with 6‐0 nylon sutures, and were classified as the ligation group. All were followed for satisfaction of penile morphology and assessed by the abridged 5‐item version of the international index of erectile function (IIEF‐5) scoring for erectile capability. In the electrocoagulation group, a mean preoperative IIEF‐5 score of 22.5 ± 1.6 decreased to a mean postoperative IIEF‐5 score of 17.9 ± 4.1. Among them 2 men (28.6%) had sustained postoperative infection. Follow‐up cavernosograms showing relatively poor filling are commensurate with intracavernosal fibrosis. In the ligation group, however, the mean IIEF‐5 score was 22.3 ± 1.9 preoperative and 22.9 ± 2.0 postoperative. Although there was no statistical significance between the 2 groups in preoperative IIEF scores, there was a significant difference between groups postoperatively. Application of electrocoagulation appears to be disadvantageous in preserving erectile tissues. A Bovie effect should be avoided in this erectile organ in order to preserve erectile capability and avoid infection.
The Journey of a Trainee
Authors: Paduch Darius A.
Published: December 2015

As you are reading this issue, you should all be making plans to attend the American Society of Andrology's 30th Annual Meeting, to be held in Seattle, Wash, from March 30 to April 6, 2005. This meeting always provides the perfect opportunity to mingle with fellow trainees and to network with other andrology professionals. This issue's segment showcases Darius Paduch's own unique journey as a trainee. We encourage you to approach him and any of the other authors who have shared their experiences and wisdom in Trainee Page articles. These individuals are ideal resources and are always happy to answer any questions that you may have regarding your career goals and plans for the future.
Effect of Vasectomy on Sperm Nuclear Chromatin Condensation in the Rabbit
Authors: Steger Klaus , Slavov Milen , Failing Klaus , Weidner Wolfgang , Bergmann Martin
Published: December 2015

ABSTRACT: Histone‐to‐protamine exchange in haploid spermatids is known to play a central role for male fertility. The present study investigates, for the first time, the effects of vasectomy on the expression of protamines in the rabbit. During normal spermatogenesis, protamine‐1 and protamine‐2 mRNA were expressed from step 5 round spermatids to step 11 elongated spermatids. In unilaterally vasectomized animals, control testes revealed normal spermatogenesis with normal protamine expression, while vasectomized testes exhibited both normal spermatogenesis and spermatogenic arrest. Some testes with normal spermatogenesis revealed delayed expression of both protamine‐1 and protamine‐2. Furthermore, multinucleated round spermatids were a regular finding in these testes. In both treated and untreated animals, a higher percentage of spermatozoa from the cauda epididymis had highly condensed chromatin when compared with those from the testis. The percentage of spermatozoa with highly condensed chromatin from testes and epididymides from the vasectomized side of treated animals remained unchanged from controls. As the integrity of nuclear chromatin is important for oocyte fertilization, especially in intracytoplasmic sperm injection (ICSI), where most of the natural selection mechanisms are bypassed, our data add valuable information for the treatment of infertility by ICSI, showing that vasectomy may affect nuclear chromatin integrity of testicular spermatids but not epididymal spermatozoa. Microsurgical epididymal sperm aspiration (MESA), therefore, may be superior to testicular sperm extraction (TESE) in vasectomized patients.
Religious Coping, Coping Resources, and Depressive Symptoms: Test of a Mediation Model
Authors: Davis Don E. , Ashby Jeff , McElroy Stacey , Hook Joshua
Published: December 2015

This study examined the relationships between religious coping, coping resources, and depressive symptoms. The authors tested whether coping resources explained the link between religious coping and depressive symptoms in a sample of 349 college students. Results indicated that coping resources partially mediated the relationship between negative religious coping and depressive symptoms, primarily through decreased social ease. The results offered no evidence that coping resources mediated the relationship between positive religious coping and depressive symptoms.