bluuebunny/crossref_metadata_2025 huggingface dataset:
Seizures in Patients Receiving Concomitant Antimuscarinics and Acetylcholinesterase Inhibitor
Authors: Piecoro Lance T. , Wermeling Daniel P. , Schmitt Frederick A. , Ashford J. Wesson
Published: October 2023

Seizures occurred in two patients with probable Alzheimer's disease who were receiving long‐term treatment with metrifonate, an irreversible acetylcholinesterase inhibitor. In both patients seizures were associated with discontinuation of short‐term agents with high antimuscarinic properties. Hence, abrupt discontinuation of antimuscarinics or anticholinergics with high antimuscarinic properties in patients receiving long‐term acetylcholinesterase inhibition therapy may be associated with a reduction of seizure threshold. With increasing administration of acetylcholinesterase inhibitors for patients with Alzheimer's disease, practitioners should be aware of the potential for drug‐drug interactions and other complications. In general, it is good medical practice to avoid concomitant administration with centrally acting anticholinergic agents.
Another Report of Adverse Reactions to Immediate‐Release Nifedipine
Authors: Fami Mohammad J. , Ho Nancy T. , Mason Carol M.
Published: October 2023

Numerous case reports in the literature describe adverse drug events associated with immediate‐release nifedipine (IRN). In addition, several publications alerted health care professionals regarding the agent. However, it is still administered, and adverse reactions are still reported. Our patient suffered IRN‐induced hypotension, myocardial ischemia, and mental status changes. We recommend that the use of IRN in treatment of hypertension should be either totally prohibited or severely restricted. The product should not be readily available for indiscriminate administration. Also, continuing education and training are required to alert all health care professionals to the serious dangers associated with this drug.
Semen Donor Selection by In Vitro Sperm Mobility Increases Fertility and Semen Storage in the Turkey Hen
Authors: DONOGHUE ANN M. , HOLSBERGER DENISE R. , EVENSON DONALD P. , FROMAN DAVID P.
Published: October 2023

ABSTRACT: Commercial turkey production relies on the artificial insemination (AI) of pooled semen. However, semen quality ultimately depends on the quality of individual ejaculates. The purpose of this study was to evaluate semen from individual toms by means of an objective sperm‐mobility assay. Semen was then pooled by mobility phenotype and inseminated into hens, and the percentages of fertile and hatched eggs were determined after egg incubation. To indirectly evaluate hens' sperm storage, we determined the number of sperm holes in the perivitelline layer (PL) of freshly laid eggs. Semen from individual ejaculates (two trials, total of 169 toms) was evaluated by use of the sperm‐mobility test (SMT). Semen was diluted to 1 × 109 sperm/ml, in prewarmed N‐tris‐[hydroxymethyl] methyl‐2‐amino‐ethanesulfonic acid (TES)‐buffered saline, and was placed over 3 ml of a 2% (w/v) Accudenz solution at 41° C. After a 5‐minute incubation period, the cuvette was placed in a densimeter, and percentage transmission was recorded after 1 minute. Semen samples from toms ranked, according to sperm mobility, in the highest 10% and the lowest 10%, after three evaluations, were pooled by group and were used to inseminate hens weekly (trial 1: n = 20 hens/group, for 10 weeks, AI dose = 150 × 106 spermatozoa inseminated fresh and after 24‐hour in vitro storage at 5° C; trial 2: n = 60 hens/group, for 16 weeks, AI dose = 75 × 106 spermatozoa inseminated fresh). Each week, eggs from day 6 post‐AI were evaluated for holes in the PL, vestiges of acrosomal induced hydrolysis. Spermatozoa from toms of different mobility phenotypes were also evaluated individually, for sperm chromatin structure and motility variables, by use of the Hobson Sperm Tracker. Toms characterized by high and low in vitro sperm‐mobility phenotype were categorized as “high mobility” and “low mobility,” respectively. The percentage of fertile eggs from hens inseminated with semen from the high‐mobility toms was higher than the percentage of fertile eggs from the low‐mobility group, in each trial (95.8 ± 1.3% vs. 90.4 ± 2.2%, and 88.7 ± 4.0% vs. 82.4 ± 0.4%, trials 1 and 2, respectively; P < 0.05). More sperm holes were observed in the PL of eggs fertilized by the high‐mobility toms than in the PL of eggs fertilized by the low‐mobility toms (P < 0.05). No differences in susceptibility of sperm nuclear DNA to denature in situ, as measured by the flow‐cytometric sperm chromatin‐structure assay, were detected between toms of differing mobility phenotypes. Sperm‐motility variables, straight‐line velocity, and average‐path velocity were significantly greater for high‐mobility toms compared to low‐mobility toms (P < 0.05). Sperm‐mobility differences between toms (detected by means of the SMT) influenced sperm storage, as indicated by the number of sperm in the PL and by the percentage of fertile eggs produced.
Hormonal Regulation of Spermatogenesis in the Hypophysectomized Rat: Cell Viability After Hormonal Replacement in Adults After Intermediate Periods of Hypophysectomy
Authors: SHENNAWY AHMED EL , GATES ROBERT J. , RUSSELL LONNIE D.
Published: October 2023

ABSTRACT: A quantitative analysis of germ‐cell populations in normal, hypophysectomized (Hx), and Hx‐hormone—treated animals was undertaken during periods of regression that were characterized as intermediate, between short‐term and long‐term regression of the testis. Twenty‐one groups of adult rats were administered either follicle‐stimulating hormone (FSH), growth hormone, thyroid‐stimulating hormone (TSH), or testosterone (T) in various doses and combinations. The dosage of T administered was less than that expected to achieve maximum testis weight. Flutamide and Casodex were used to compete with androgen binding to receptors in Hx animals, as it is known that small amounts of androgen are secreted in the absence of pituitary stimulation. Follicle‐stimulating hormone, T, and TSH all significantly maintained testis weight as compared with Hx controls, although FSH and T, singly or in combination, were the most effective. Contamination of the TSH preparation with trace amounts of FSH was apparently responsible for the slight maintenance of testis weight. A novel assay for determination of the numbers of viable germ cells was used in a subset of these groups to determine the cellular sites of FSH and T action. Numbers of type A spermatogonia were lowered after Hx and were maintained by either FSH or T or a combination of these hormones. Other phases of germ‐cell development most susceptible to FSH and/or T were the successive conversions of type A spermatogonia to intermediate spermatogonia, intermediate spermatogonia to type B spermatogonia, preleptotene spermatocytes to pachytene spermatocytes, and early pachytene spermatocytes to intermediate maturity pachytene spermatocytes during early and midcycle phases of pachytene spermatocyte development. Germ‐cell loss during meiosis and virtually every phase of spermatid development was largely prevented by FSH or T or a combination of these hormones. Thus, in testes in advanced stages of regression, both FSH and T were capable of preventing cell loss, suggesting that both hormones can affect the survival of the same cell type. The present study demonstrated that FSH can partially compensate for lowered T levels. The combined administration of FSH and T was more effective in preventing overall cell degeneration than either hormone alone. Unlike the initial phase of spermatogenesis, in which there is a largely midcycle loss of germ cells due to Hx, the loss of cells during testis regression is more widespread and impacts several cell types in more than one stage of the spermatogone cycle.
Hormonal Regulation of Spermatogenesis in the Hypophysectomized Rat: Quantitation of Germ‐Cell Population and Effect of Elimination of Residual Testosterone After Long‐Term Hypophysectomy
Authors: FRANCA LUIZ R. , PARREIRA GLEYDES G. , GATES ROBERT J. , RUSSELL LONNIE D.
Published: October 2023

ABSTRACT: Spermatogenesis continues after long‐term hypophysectomy (Hx), but massive cell degeneration prevents seminiferous tubules from attaining the full complement of cells. One objective of this study was to determine the vulnerable sites for completion of spermatogenesis in long‐term Hx rats. It is now known that Leydig cells continue to secrete small amounts of androgen after Hx. A second objective was to determine the cellular sites that are maintained by residual androgen secreted by Leydig cells post‐Hx. Two groups of adult animals were utilized. Both groups were Hx for 36 days, but one group of rats received the androgen antagonist flutamide during the 26th through the 36th day of Hx (10 days). Germ‐cell numbers were quantified using a method that allowed their expression as numbers of cells present per hour of development. In the long‐term Hx rat, the germ‐cell population increased to preleptotene, but the divisions that led to preleptotene were inefficient due to cell degeneration. Subsequent to preleptotene, there was a gradual loss in cells such that there were few germ cells remaining by steps 9–13. Flutamide given to Hx rats did not result in a significant difference in the numbers of intermediate and type B spermatogonia or significant differences in progenitor cells. A significant and major depression of cell numbers in Hx‐flutamide‐treated rats occurred in the cell division of type B spermatogonia to form preleptotene spermatocytes. There was a less dramatic, although significant, depression of cell numbers in Hx‐flutamide—treated animals that occurred from preleptotene until late pachytene as well as an increased loss of round spermatids at midcycle (step 5–6). These data demonstrate that cell loss after long‐term Hx occurs at numerous phases of spermatogenesis. The data also demonstrate that the presence of residual androgen action after long‐term Hx results in enhanced germ‐cell survival. Although the major blockage in cell viability occurs at midcycle steps in the long‐term Hx rat, there are several other hormone‐sensitive phases of spermatogenesis.
Seminal Vesicles Are Novel Sites of Luteinizing Hormone/Human Chorionic Gonadotropin—Receptor Gene Expression
Authors: TAO Y.‐X. , LEI Z. M. , RAO Ch. V.
Published: October 2023

ABSTRACT: The hypothesis that rat seminal vesicles may contain luteinizing hormone (LH)/human chorionic gonadotropin (hCQ) receptors was tested by means of a number of different techniques. Northern blotting demonstrated that rat seminal vesicles contained multiple LH/hCG‐receptor transcripts. In situ hybridization revealed that these transcripts were present primarily in the principal epithelial cells lining the lumen. Western immunoblotting detected proteins, two of which disappeared (80 and 46 kDa) and another of which decreased (30 kDa) after preabsorption of the receptor antibody with excess receptor peptide. Ligand blotting showed that 125I‐hCG could bind only to ah 80‐kDa protein and that this binding was inhibited by coincubation with excess unlabeled hCG. Immunocytochemistry demonstrated that LH/hCG‐receptor protein was present primarily in the principal epithelial cells. In conclusion, our data demonstrate that seminal vesicles contain LH/hCG receptors, thus making this previously unsuspected male accessory reproductive organ a potential target of direct regulation by LH.
Ethnic Differences in Testicular Structure and Spermatogenic Potential May Predispose Testes of Asian Men to a Heightened Sensitivity to Steroidal Contraceptives
Authors: JOHNSON LARRY , BARNARD JEFFREY J. , RODRIGUEZ LORI , SMITH ELIZABETH C. , SWERDLOFF RONALD S. , WANG X. H. , WANG CHRISTINA
Published: October 2023

ABSTRACT: Spermatogenesis in Asian men appears to be more susceptible to suppression by steroidal contraceptives administered in clinical trials than spermatogenesis in Caucasian men. The objective of this study was to determine whether ethnic differences exist in testicular structure and spermatogenic potential that might predispose Asians to a high sensitivity to steroidal contraceptives. Testes from 12 Chinese men were compared to those from 8 Hispanic men and 12 non‐Hispanic Caucasian men of ages 29 ± 3, 30 ± 2, and 29 ± 3 years, respectively. Testes were fixed by vascular perfusion with glutaraldehyde, further fixed in osmium, embedded in Epon, and evaluated by stereology using 0.5‐μm sections stained with toluidine blue. Homogenates of fixed testes were evaluated for the number of Sertoli cells and the daily sperm production based on pachytene primary spermatocytes (PDSP) or spermatids with spherical nuclei (DSP). Paired parenchymal weight was less (P < 0.05) in Chinese men than in Hispanic or Caucasian men. The PDSP per gram of parenchyma was lower (P < 0.05) and the DSP per gram tended to be lower in Chinese men than in other groups. The histologic appearance, volume density, and length per man of seminiferous tubules were the same among the ethnic groups; however, the diameter of seminiferous tubules was less (P < 0.05) in Chinese than in Hispanic or Caucasian men. The PDSP per man and the DSP per man were lower (P < 0.05) in Chinese than in Hispanic or Caucasian men. The number of Sertoli cells per gram was higher (P < 0.05) in Chinese or Caucasian men than in Hispanic men, but the number of Sertoli cells per man was lower (P < 0.05) in Chinese men than in Hispanic or Caucasian men. Sertoli cell function, measured as the number of germ cells accommodated by a single Sertoli cell, was lower (P < 0.05) in Chinese men than in Caucasian men. The volume density of Leydig cell cytoplasm was greatest (P < 0.05) in Chinese men, but the number of Leydig cells was similar among the ethnic groups. Hence, smaller testes coupled with reduced Sertoli cell number and function and reduced daily sperm production could predispose Asian men to have a heightened negative response of testes to steroidal contraceptives, as compared to Caucasian men. Dampening (by exogenous androgens) of any physiological benefit to spermatogenesis that a high volume density of Leydig cell cytoplasm may bestow on the human testis (that Asian men may have evolved to require) would exacerbate ethnic differences in the spermatogenic response to hormonal contraceptives.
Neuronal Nitric Oxide Synthase in the Canine Prostate: Aging, Sex Steroid, and Pathology Correlations
Authors: CRONE JULIE K. , BURNETT ARTHUR L. , CHAMNESS SHELLY L. , STRANDBERG JOHN D. , CHANG THOMAS S. K.
Published: October 2023

ABSTRACT: Nitric oxide synthase (NOS) is expressed in the prostate of various species, including humans. NOS catalyzes the production of nitric oxide (NO), which may function in prostatic smooth‐muscle relaxation. To investigate further the role of NO in the prostate, we examined neuronal NOS expression in the aging canine prostate, after hormonal perturbation, and correlated these results with histopathologic findings. The study comprised the following treatment groups: intact dogs (treatment group 1, n = 6); dogs who were castrated at 7 days of age and received testosterone and estrogen replacement at 2 years of age (treatment group 2, n = 10); and dogs who were castrated at 2 years of age and received testosterone and estrogen replacement at 2 years of age (treatment group 3, n = 9). Studies were done on prostates removed from dogs after euthanasia at 6 years of age. In treatment group 1, complex benign prostatic hyperplasia (BPH) was observed in all specimens. In treatment group 2, atrophy was observed in 70%, normal prostate with small areas of hyperplasia in 20%, and BPH in 10% of specimens. In treatment group 3, atrophy was observed in 78%, normal histology with small areas of hyperplasia in 11%, and BPH in 11% of specimens. Neuronal NOS localizations were confirmed by western blot analysis and by immunohistochemistry in 0% and 17%, respectively, of specimens in treatment group 1, in 60% and 70%, respectively, of specimens in treatment group 2, and in 67% and 71%, respectively, of specimens in treatment group 3. Neuronal NOS immunoreactivity was localized in histologically normal prostates of four intact, young‐adult control dogs (2 years of age). For all treatment groups, neuronal NOS immunoreactivity was confirmed by western blot in 86% of atrophic prostates but in no prostates with BPH (P < 0.001), and it was confirmed by immunohistochemistry in 75% of atrophic prostates but in only 13% of prostates with BPH (P < 0.02). These data suggest that, in the canine prostate, NO release relates to growth and pathology. Low levels of neuronal NOS expression in BPH tissue, compared with higher levels in atrophic tissue, suggest that neuronal NOS expression is down‐regulated in the prostate with benign cellular proliferation whereas it is maintained or possibly up‐regulated in the prostate with prostatic involution. Whether altered neuronal NOS expression contributes to the pathogeneses of BPH and prostatic involution or whether it occurs as a consequence of these processes requires further investigation.
SP22: A Novel Fertility Protein From a Highly Conserved Gene Family
Authors: WELCH JEFFREY E. , BARBEE RANDY R. , ROBERTS NAOMI L. , SUAREZ JUAN D. , KLINEFELTER GARY R.
Published: October 2023

ABSTRACT: Three nucleotide sequences encoding SP22, a protein originally identified in detergent extracts of cauda epididymal sperm, were isolated from a rat testis cDNA library. While two of these cDNA sequences differed only in their 5′ untranslated regions, a third cDNA was predicted to contain an additional 13 amino acids of coding sequence. Amino acid sequences obtained following Edman degradation of purified SP22 protein and cDNA sequence data both indicated that SP22 was a member of a highly conserved and widely expressed gene family found in organisms as diverse as human and Escherichia coli. Interestingly, while a 1‐kb mRNA transcript was widely expressed in somatic tissues, a unique pattern of testicular expression was observed, including the appearance of a novel 1.5‐kb transcript and an increase in the abundance of the 1‐kb transcript during spermatogenic cell development. Anti‐SP22 peptide antiserum was shown to recognize a family of 22‐kDa proteins on western blots of detergent‐extracted cauda epididymal sperm protein, suggesting that multiple charge variants of SP22 coexist. Moreover, affinity‐purified anti‐SP22 peptide immunoglobulin localized in a highly specific manner to the anterior‐ventral surface of the equatorial segment of the sperm head. This is an extremely intriguing finding as SP22 was originally shown to be highly correlated with, and predictive of, the fertilizing ability of cauda epididymal sperm. Although no conclusive function has been attributed to any members of the SP22 gene family, the localization of SP22 over a discrete region of the sperm head suggests a pivotal role in sperm—egg interactions.
Deficiency of Fucosidase Results in Acrosomal Dysgenesis and Impaired Sperm Maturation
Authors: VEERAMACHANENI D. N. RAO , SMITH MARY O. , ELLINWOOD N. MATTHEW
Published: October 2023

ABSTRACT:Although a variety of glycosyltransferases and glycosidases have been implicated in spermatogenesis and posttesticular sperm maturation, the biological role of these enzymes in these processes is largely unknown. We describe reproductive sequelae in a cohort of male dogs suffering from fucosidosis, a heritable lysosomal storage disorder caused by a severe deficiency of α‐l‐fucosidase. There was a reduction in the total number of sperm in the ejaculate. Only 3–5% of sperm were motile. None of the sperm were found to be morphologically normal. The predominant morphological defects observed were malformed acrosomes (56%) and retained proximal cytoplasmic droplets (92%), indicating that spermiogenesis and sperm maturation were impaired. The cytoplasm of all cellular components of the testis and excurrent ducts were vacuolated. The vacuolation resulted from enlargement of lysosomes caused by accumulation of compounds that are otherwise cleaved/degraded when lysosomal hydrolases are present normally. It is possible that impairment in spermatogenesis, particularly morphogenesis of the acrosome, is due to physical damage caused by anomalous enlargement of lysosomes. Although an unambiguous causal relationship could not be established, it is evident from the available information that the derangement in events associated with epididymal sperm maturation, namely acquisition of motility and shedding of the cytoplasmic droplet, is likely due to lack of fucosidase leading to impaired sperm membrane modification. This heritable condition in dogs may serve as a spontaneously occurring knock‐out model for further elucidating the role of α‐l‐fucosidase in spermatogenesis and sperm maturation.
The Effects of Aging on the Expression of Glutathione S‐Transferases in the Testis and Epididymis of the Brown Norway Rat
Authors: MUELLER A. , HERMO L. , ROBAIRE B.
Published: October 2023

ABSTRACT: Glutathione S‐transferases (GSTs), a family of isoenzymes, catalyze the conjugation of glutathione to a variety of electrophiles, and protect cellular constituents from electrophilic and oxidative attack. Aging is associated with an overall increase in oxidative stress and thus free radical production. The present study examines the Immunocytochemical localization of Ya, Yc, Yb1, Yb2, Yo, and Yf GST subunits in the testis and epididymis of Brown Norway rats aged 3, 12, 18, and 24 months. In the testis, neither Sertoli nor germ cells showed changes in the GST staining pattern during aging. At 24 months, two types of Leydig cells were noted. Some (peritubular) formed a distinct band at the periphery of the tubule while others were seen in the interstitial space. The peritubular cells were identified as Leydig cells by specific staining for 3β‐hydroxysteroid dehydrogenase (3β‐HSD), a Leydig cell—specific marker. Both types of Leydig cells were intensely reactive for all GST subunits at all ages. In the epididymis, principal cells of all epididymal regions, except the proximal cauda region, showed no changes in GST expression at all ages examined. At 24 months, some principal cells of this region became greatly enlarged and vacuolated. These cells were unreactive for Yo, Yb1, Yb2, and Yc, while adjacent normal‐appearing principal cells maintained the same intensity of expression as seen in 3‐month controls. In contrast, vacuolated principal cells were reactive for the Ya subunit, while adjacent normal principal cells were unreactive. These data indicate that selective changes occur in the expression of GSTs at 24 months in principal cells having both a normal and a vacuolated appearance. The underlying mechanism responsible for these changes with age is unresolved, but we speculate that they lose the ability to handle oxidative stress. Taken together, these data show that aging affects region‐specific changes in GST expression in the epididymis and Leydig cell distribution in the testis.
The Loss of α‐Adrenergic Effect during the Erectile Response in the Long‐Term Diabetic Rat
Authors: MILLS T. M. , LEWIS R. W. , STOPPER V. S. , REILLY C. M.
Published: October 2023

ABSTRACT: The present study was designed to investigate the effect of long‐term, streptozotocin‐induced diabetes on the erectile response in the laboratory rat. Mean arterial blood pressure (MAP) and intracavernosal blood pressure within the erectile tissue (CCP) were continuously monitored during erection elicited by stimulation of the autonomic innervation of the penis. MAP and CCP were also measured during administration of two drugs: nitroglycerin, a nitric oxide donor drug and phenylephrine, an α‐adrenergic agonist. The results of these studies show that during graded electrical stimulation of the ganglion, the overall magnitude of the erectile response was greater in the diabetic rats than in untreated control animals. Neither diabetic nor control animals responded significantly to infusion of nitroglycerin. However, diabetic rats and control rats responded very differently to administration of phenylephrine; in the control rats, this a agonist caused a sharp decline in CCP as the cavemosal vessels constricted in response to the drug. The same dose of phenylephrine had no discernible effect on CCP in the diabetic animals. This loss of α responsiveness may be confined to the penile circulation because MAP was elevated to approximately the same extent in both groups. Taken together, these results show that long‐term diabetes leads to a failure of α‐adrenergic responsiveness in the cavemosal circulation. The greater erectile response to ganglionic stimulation in the diabetic animals is likely due to the loss of response to endogenous norepinephrine.
Triptolide: A Potential Male Contraceptive
Authors: LUE YANHE , HIKIM AMIYA P. SINHA , WANG CHRISTINA , LEUNG ANDREW , BARAVARIAN SIMA , REUTRAKUL VICHAI , SANGSAWAN RANEE , CHAICHANA SUTTIPORN , SWERDLOFF RONALD S.
Published: October 2023

ABSTRACT: The antifertility effect of triptolide and other related compounds, isolated from Tripterygium wilfordii, has been demonstrated in male rats. The exact sites and mechanism of action of triptolide remain unknown. Our objectives were to determine whether triptolide at selected dose levels that induce infertility has any detrimental effects on the testes and to determine the sites and the possible mechanisms of its action. Groups of six adult male Sprague‐Dawley rats were given oral administration of either vehicle (control group) or triptolide (50 or 100 μg/kg body weight) daily for 35 or 70 days. Body weight gain was normal in all treated groups. All six rats treated with a high dosage of triptolide were infertile during the second (63–70 days) mating trial. A lower dose (50 μg) of triptolide gave intermediate fertility values. Plasma levels of luteinizing hormone, follicle‐stimulating hormone, testosterone, and intratesticular testosterone were not significantly different between control and triptolide‐treated groups. Cauda epididymal sperm content was decreased by 68% and the motility, which averaged 58.2% in the control rat, was reduced to almost zero. No effects of triptolide were observed on testis and accessory organs weight, volumes of tubular lumen and the total Leydig cells, tubule diameter, and the number of Sertoli cells, spermatogonia, preleptotene (PL), and pachytene (P) spermatocytes. There were, however, modest but significant decreases in tubule volume and the number of round spermatids at stages VII—VIII. No changes in the germ cell apoptotic index measured at stages VII—VIII and XIV—I were noted between controls and rats rendered infertile with a high dose of triptolide. Thus, triptolide, at a dose level that induces complete infertility in the adult rats, has minimal adverse effects on the testes and acts primarily on the epididymal sperm making triptolide an attractive lead as a post‐testicular male contraceptive.
Apoptosis Pattern Elicited by Several Apoptogenic Agents on the Seminiferous Epithelium of the Adult Rat Testis
Authors: BLANCO‐RODRÍGUEZ JOSEFA , MARTÍNEZ‐GARCÍA CARMEN
Published: October 2023

ABSTRACT: Spontaneous germ cell death during spermatogenesis is an important event, and the usefulness of the seminiferous epithelium as an in vivo model to study apoptosis has been evidenced. Nevertheless, the response of the testis to apoptogenic agents has not been analyzed. This study was designed to determine germ cell sensitivity to induction of apoptosis and to provide baseline data on the testis response to several apoptogenic agents. Induced apoptosis was assessed by in situ DNA 3′‐end labeling and quantified at every stage of the spermatogenic cycle. The shortest response time for every agent was established based on morphological and quantitative criteria. Our results show significantly increased incidence of germ cell deaths after all treatments, mainly at stages I, XII, and XIV. These specific stages coincide with those at which the greatest numbers of spontaneous germ cell deaths occur in control animals. Moreover, the rapid and highly specific response of germ cells to all the apoptogenic agents used in the present study indicate that apoptosis must be tightly regulated at these stages of the seminiferous epithelium. As a consequence, we propose that the disruption of apoptosis control might be an important determinant for idiopathic male infertility.
Androgen Binding Protein Secretion and Endocytosis by Principal Cells in the Adult Rat Epididymis and During Postnatal Development
Authors: HERMO LOUIS , BARIN KATY , OKO RICHARD
Published: October 2023

ABSTRACT: Androgen binding protein (ABP) has been shown to be secreted by Sertoli cells and to be actively taken up by the efferent ducts and proximal caput epididymidis and, yet, to be present at high concentrations in epididymal fluids. In the present study, ABP was immunolocalized by light microscopy in epithelial cells of the efferent ducts and epididymis of adult rats and during postnatal development and by electron microscopy in specific organelles within these cells. In adults, the efferent ducts actively endocytosed Sertoli cell‐derived ABP. In the epididymis, principal cells displayed a variable staining reminiscent of a checkerboardlike pattern, with cells being intensely, moderately, or weakly reactive throughout their cytoplasm or unreactive. In the electron microscope, reactive cells displayed a labeling of their Golgi apparatus and secretory vesicles indicative of an epididymal‐secreted form of ABP. However, labeling was also noted over endosomes of principal cells, but only of the initial segment and intermediate zone, which, along with labeling of coated pits and vesicles, indicated that ABP was also endocytosed by principal cells of these regions. The postnatal study revealed that principal cells attained an adultlike staining pattern indicative of secretion in a region‐specific manner at different ages, suggesting that ABP secretion is regulated by different factors. Ligation of the etferent ducts of 15‐day‐old animals revealed no reaction along the entire epididymis in animals sacrificed at later ages, suggesting the importance of luminal testicular factors in its regulation during development. In addition, as in the adult, ABP was also endocytosed by principal cells, but only in the initial segment and intermediate zone. Taken together, the present results indicate that secretion of ABP occurs along the entire epididymis, whereas endocytosis is region specific. The functional role of ABP in the epididymis in relation to sperm maturation is discussed.
Selective Expression of a Glutathione S‐Transferase Subclass During Spermatogenesis
Authors: ROWE JONATHAN D. , TCHAIKOVSKAYA TATYANA , SHINTANI NAOAKI , LISTOWSKY IRVING
Published: October 2023

ABSTRACT: Levels of the hGSTM3 glutathione S‐transferase (GST) subunit in testis of the human fetus and infant were found to be only a small fraction of those in adults. To understand these observations and to determine whether hGSTM3 subunit expression is developmentallly and/or hormonally regulated, an experimental model based on the rat testis homologue (subunit rGSTM5) was used. For prepubertal rats, testicular rGSTM5 subunit levels were very low, but a sharp increase was observed between weeks 6 and 7 of development, when testicular growth includes increased numbers of germ cells associated with spermatogenesis. In adult hypophysectomized rats, the rGSTM5 subunit content of testis decreased progressively over 5 weeks, at which time the subunit was barely detectable. In contrast, the other GST subunit types did not vary significantly during development or after hypophysectomy. These results suggest that rGSTM5 subunits in rat testis could originate from spermatogenic cells. Accordingly, GSTs were purified from human sperm, and it was shown that the hGSTM3 subunit was, by a large measure, the predominant form. These data are consistent with the notion that the differential expression of hGSTM3 during human testicular development can also be explained on the basis of its preferential location in germs cells.
Unilateral Injection of Neuropeptide Y Decreases Blood Flow in the Injected Testis but May Also Increase Blood Flow in the Contralateral Testis
Authors: COLLIN OLA , ENFÄLT ELIN , ÅSTRÖM MAGDALENA , LISSBRANT ERIK , BERGH ANDERS
Published: October 2023

ABSTRACT: Neuropeptide Y (NPY) receptors have recently been described in intratesticular arterioles, but the role of NPY in testicular blood‐flow regulation has not been examined. To explore this, we administered NPY in various doses (0.01–10 μg) via intratesticular injections and studied testicular microcirculation using a laser Doppler flow meter. NPY injection shows a dose‐response pattern, with 1 μg (the most potent dose) causing a decrease (−42.4 ± 3.7%, P 0.00005) in blood flow in the ipsilateral testis of all the animals and an increase in blood flow in the contralateral testis (+17.2 ± 5.6%, P = 0.03, n = 25 animals). The response in the contralateral testis was variable. A clear‐cut increase was seen in 19 of the 25 animals examined, whereas either no response or a slight decrease was seen in the remaining six. The contralateral increase, which was not seen in the hindpaw on the same side, did not occur when the neuronal connections to the testes were blocked by injection of local anesthetics into the spermatic cord, either on the NPY‐injected side or on the contralateral side. Our results suggest that NPY may serve as a vasoconstrictor in the testis, probably by acting on the NPY‐Y1 receptors present on intratesticular arterioles. Local injection of NPY causes a major decrease in blood flow in the injected testis. This decrease is followed in the majority of animals studied by an increase in blood flow in the contralateral testis, an effect that seems to depend on neuronal mechanisms. This observation suggests that the testes may communicate under certain situations. The functional consequences of this remain to be elucidated.
Bicarbonate‐Dependent Lipid Ordering and Protein Aggregation Are Part of the Nongenomic Action of Progesterone on Capacitated Spermatozoa
Authors: PUROHIT SHARAD B. , LALORAYA MALINI , KUMAR PRADEEP G.
Published: October 2023

ABSTRACT: The nongenomic action of progesterone (P) on capacitated sperm In mediating acrosomal exocytosls operates through transmembrane signal transduction involving Increased Intracellular calcium Ions and modulation of protein Kinases and phospholipid metabolism through a second messenger pathway. Conflicting views exist regarding the nature of the nongenomic receptor of P. It Is thought to be a cellsurface receptor having the properties of a calcium channel, chloride channel, bicarbonate/chloride exchanger, and gamma amlnobutyrtc acid type A (GABAA. In this study, we tried to understand the role of bicarbonate and/or P in inducing membrane perturbations in capacitated and acrosome‐reacting spermatozoa. We also attempted to characterize the membrane responses in P‐stimulated and bicuculline‐poisoned sperm. The presence of a high level of bicarbonate in the medium favored high rotational mobility of lipids and proteins in capacitated sperm. However, the capacitated sperm exhibited low lipid ordering and unaltered protein ordering in response to the high bicarbonate concentration. P and W‐cuculline appeared to bind to the same or similar binding sites, probably a GABAA receptor, eliciting similar motional perturbations In sperm membranes. It appears that both P and bteuculllne can Induce receptor aggregation and lipid ordering In sperm membranes, which alter the capability of sperm to bind zona pelludda, and that the P action on sperm takes effect through a bicarbonate‐dependent signal transduction mechanism.
Expression of Cydin‐Dependent Kinase 5 and Associated Cyclins in Leydig and Sertoli Cells of the Testis
Authors: MUSA FATHEL RAHMAN M. , TOKUDA MASAAKI , KUWATA YOSHIHIRO , OGAWA TAKAAKI , TOMIZAWA KAZUHTTO , KONISHI RYOJI , TAKENAKA IKUMASA , HATASE OSAMU
Published: October 2023

ABSTRACT: In this study, we examined the expression and subcellular localization of cyclin‐dependent kinase 5 (Cdk5), cyclin D1, and cyclin E in Leydig and Sertoli cell lines that were cultured with 7.5, 1.0, 0.5, or 0% serum (mixture of a 2:1 ratio of horse serum and fetal bovine serum) and in the developing rat testis to verify the possible functions of Cdk5, cyclin D1, and cyclin E in the testis. The abundance of Cdk5 and cyclin E in the Leydig cell line, TM3, was significantly reduced at low serum concentrations. In contrast, serum concentration had no effect on Cdk5 and cyclin E levels in the Sertoli cell line, TM4.

Cyclin D1 was detected by western blot analysis in TM4 cells only, and its abundance was serum dose dependent. The kinase activity of Cdk5 in TM3 and TM4 cells that were cultured at various serum concentrations coincided with the levels of Cdk5 expression. Immunohistochemical staining for Cdk5 and cyclin E reealed nuclear and cytoplasmic distribution, both in TM3 and TM4 cells. Moreover, cyclin D1 immunoreactivity was only detected in TM4 cells. In the developing rat testis, Cdk5 expression was most prominent at 2 and 3 weeks after birth. Cyclin D1 was strongly expressed at 1 and 2 weeks in premature rat testes. On the other hand, cyclin E was highly expressed in the adult testis. Immunohistochemical localization of Cdk5, cyclin D1, and cyclin E in 1‐week‐old and adult rat testes revealed expression in both Leydig and Sertoli cells. Our results suggest that Cdk5 in TM3 and Leydig cells of the testis might play a role in cell cycle regulation, whereas Cdk5 in TM4 and Sertoli cells of the adult testis might have some additional functions besides control of proliferation. Key words: Kinase activity, growth control.
Sperm Motility Enhancement by Nitric Oxide Produced by the Oocytes of Fathead Minnows, Pimephelas promelas
Authors: CREECH MARGARET M. , ARNOLD ERNST V. , BOYLE BRIAN , MUZENICH MICHAEL C. , MONTVELLE CHRISTOPHER , BOHLE D. SCOTT , ATHERTON ROBERT W.
Published: October 2023

ABSTRACT: The effects of nitric oxide (NO) on sperm motility were examined in the fathead minnow, Pimephelas promelas, using computer‐assisted sperm analysis (CASA). The observed effects underscore the dual nature of NO as both a low‐concentration regulatory agent and, at higher doses, a cytotoxic agent. At 1 × 10∼6 M concentration, NO donor sodium nitroprusside (SNP) enhanced sperm motility percentages and increased CASA velocity parameters curvilinear velocity, straight‐line velocity, and average path velocity, whereas 1 × 10−2 M concentration inhibited percent motility and decreased velocities. Fathead minnow ova‐produced NO was subsequently trapped as a paramagnetic ferrous iron complex and detected by electron spin resonance spectroscopy. The distinctive triplet spectrum, with aN = 12.5G and giso = 2.04, was recorded during a critical 5‐minute period following laying. Nitric oxide synthase (NOS) was histochemically localized at the micropyle of mature unfertilized fathead eggs, and an inhibitor of NOS blocked histochemical staining. CASA analysis of sperm motility in the presence of ova‐produced NO over an 8‐minute time course reveals an optimum motility enhancement at 4 minutes that is similar to the effect of 1 × 10−6 M SNP. This transient NO production by freshly laid ova and the localization of NOS near the site of sperm entry, together with the motility‐enhancing effect of 1 × 10−6 M SNP on sperm, indicates an active role for low‐concentration NO in fertilization.
Effects of Cryopreservation on Bull Sperm Head Morphometry
Authors: GRAVANCE CURTIS G. , VISHWANATH R. , PITT COLIN , GARNER DUANE L. , CASEY PATRICK J.
Published: October 2023

ABSTRACT: Artificial insemination using cryopreserved semen is a common management tool of the contemporary livestock producer. However, cryopreservation is detrimental to sperm function and fertility, killing some 50% of the spermatozoa during the process. Prediction of cryopreservation damage from prefreeze samples remains elusive. Computer‐automated sperm head morphometry was used in this study to determine the effects of cryopreservation on bovine sperm head morphometry.

Semen was collected from 18 bulls and was divided. One portion was extended to 200 × 106 sperm/ml and a microscope slide was prepared, while the remaining portion was cryopreserved in a Triscitrateyolk extender. After thawing, the cryopreserved samples were prepared on microscope slides. All slides were air dried and were stained with hematoxylin and rose bengal. The morphometry dimensions for length, width, width/length, area, and perimeter for a minimum of 200 sperm heads were analyzed from each slide by computer‐aided sperm head morphometry analysis, and the mean measurements were recorded. Bull sperm heads were significantly I (P < 0.01) smaller in cryopreserved spermatozoa than in the companion extended samples for length (8.56 ± 0.07 vs. 8.63 ± 0.08 μm), width (4.39 ± 0.05 vs. 4.48 ± 0.05 μm), area (28.42 ± 0.07 vs. 29.14 ± 0.08 μm), and perimeter (23.33 ± 0.21 vs. 23.70 ± 0.23 μm) for all bulls. Width/length was also different (0.513 vs 0.519). In addition, differences (P < 0.05) were found within 14 of 18 bulls for at least four of the morphometric parameters. The percent change in measures after cryopreservation were correlated (P ± 0.05) to the variability of the extended sample. Variations in sperm head measurements were lower (P ≥ 0.05) in extended samples of the four bulls in which no changes occurred than in extended samples of the remaining 14 bulls. These data suggest that the variability in sperm head measurements of individual bulls, or ejaculates, may be an indicator of sperm cryosurvivability.
Extender Components and Surfactants Affect Boar Sperm Function and Membrane Behavior During Cryopreservation
Authors: PETTITT MURRAY J. , BUHR MARY M.
Published: October 2023

ABSTRACT: To determine how the individual components of extenders affected boar sperm function and membrane structure and to test a new surfactant's cryoprotective ability, boar sperm were cryopreserved in straws in BF5 extender plus or minus egg yolk plus or minus glycerol plus or minus a surfactant (Orvus ES Paste [OEP] or various concentrations of Pluronic F‐127). After thawing, sperm function and fluidity of the isolated head plasma membrane (HPM) were determined. Total motility and adenosine triphosphate content (a measure of viability) were superior postthaw in sperm extended in egg yolk plus glycerol (P < 0.05); neither surfactant improved function. Egg yolk plus any other ingredients improved normal acrosome morphology, whereas a combined measure of motility and normal acrosome morphology was better in the presence of 0.33% OEP or 0.1% Pluronic F‐127 (P < 0.05 vs. controls). Head plasma membrane was isolated from freshly collected spermatozoa and spermatozoa cryopreserved in the various extenders. Membrane fluidity was monitored with the probes cis‐parinaric acid (cPNA), transparinaric acid (tPNA), and 1,6‐diphenyl‐1,3,5‐hexatriene (DPH). The cPNA and the DPH monitor the fluidity of gel and liquid‐crystalline areas of the membrane, whereas the tPNA preferentially monitors the gel‐phase domains of the membrane. Additionally, DPH monitors the hydrophobic core of the bilayer. In the HPM from fresh sperm, the fluidity of each domain changed over time in a manner unique to that domain, and the behavior of the DPH domain varied among boars. The fluidity dynamics of each domain responded uniquely to cryopreservation. The cPNA domain was unaffected, the tPNA domain was altered by four of the eight extenders, and all extenders affected the fluidity of the DPH domain. Membrane structure was significantly correlated with cell function for sperm cryopreserved in extenders that preserved viability and motility. Sperm cryopreserved in egg yolk plus glycerol plus either OEP or 0.1% Pluronic F‐127 functioned best when the bulk domains were less fluid initially and the gel domain solidified more slowly. Therefore, the behavior of domains in the HPM of boar spermatozoa is affected by cryopreservation and is related to the postthaw function of boar sperm cryopreserved in different extenders.
[Tyr1]‐Nociceptin and Nociceptin Have Similar Naloxone Insensitive Erectile Activity in the Cat
Authors: CHAMPION HUNTER C. , BIVALACQUA TRINITY J. , WANG RUN , HELLSTROM WAYNE J. G. , KADOWITZ PHILIP J.
Published: October 2023

ABSTRACT: The heptadecapeptide nociceptin, also known as Orphanin FQ, is a newly discovered endogenous ligand for the opioidlike G‐protein‐coupled receptor ORLv The present study was undertaken to investigate responses to intracavemosal injections of the nociceptin analog [Tyr1]‐nociceptin and to investigate the effects of naloxone on erectile responses in anesthetized cats to [Tyr1]‐nociceptin and to nociceptin. Intracavemosal injections of [Tyr1]‐nociceptin and of nociceptin in doses of 0.3–30 nmol elicited dose‐related increases in cavernosal pressure, which, at the highest dose studied, were comparable to increases induced by the triple‐drug standard (papaverine, phentolamine, and prostaglandin E1), a preparation used in the treatment of erectile dysfunction. Responses to [Tyr1]‐nociceptin were rapid in onset and had a time course similar to responses to nociceptin. Metenkephalin increased cavernosal pressure, whereas injections of nociceptin‐(2–17), dynorphin A, and β‐endorphin did not alter cavernosal pressure. Erectile responses to nociceptin and to [Tyr1]‐nociceptin were not altered after administration of the opioid receptor antagonist naloxone at a time when erectile responses to metenkephalin were attenuated. These data show that [Tyr1]‐noctceptin and nociceptin have similar naloxone‐insensitive erectile activity in the cat.