Number Of Primary Spermatocytes Research Articles

The influence of ghrelin agonist ipamorelin acetate on the hypothalamic-pituitary-testicular axis in a cichlid fish, Oreochromis mossambicus

Ghrelin, a peptide found in the brain and gut, is predicted to play a significant role in the control of various physiological systems in fish. The objective of this study was to examine the impact of ipamorelin acetate (IPA), a ghrelin agonist, on the reproductive axis of the tilapia Oreochromis mossambicus. The administration of either 5 or 30 µg of IPA for 21 days led to a significant and dose-dependent rise in food intake concomitant with a significant increase in the numbers of primary spermatocytes, secondary spermatocytes, and early spermatids compared to the control group. There was a significant rise in the number of late spermatids, as well as the areas of the lobule and lumen, in fish treated with 30 µg of IPA, compared to the control group. Moreover, there was no significant difference in the percentage of gonadotropin-releasing hormone (GnRH)-immunoreactive fibres in the hypothalamus and anterior pituitary gland across different groups. However, a significant elevation in the expression of androgen receptor protein was observed in fish treated with 30 µg of IPA. Furthermore, the concentrations of luteinizing hormone (LH) and 11-ketotestosterone (11-KT) in the serum of fish treated with either 5 or 30 µg of IPA were significantly elevated in comparison to the control group. Collectively, these findings suggest that the administration of ghrelin enhances the development of germ cells during the meiosis-I phase and that this effect might be mediated via the stimulation of 11-KT and androgen receptors at the testicular level and LH at the pituitary level in the tilapia.

Synergic Effects of Rosemary Extract and Aerobic Exercise on Sperm Parameters and Testicular Tissue in an Aged Rat Model

Background: Aging, which has globally increased, reduces male fertility and damages the testes. The effect of aerobic exercises and the use of medicinal plants like rosemary on fertility remains controversial. Objectives: The aim of this study was to evaluate the effects of rosemary extract and exercise on testicular tissue. Methods: In this study, 30 male Wistar rats were divided into five groups: (1) control group (without treatment); (2) gavage group [received the solvent of rosemary extract (distilled water) and put on a turned-off treadmill for 10 minutes daily]; (3) exercise group (put on a turned-on treadmill based on an exercise protocol for 12 weeks); (4) extract group (received 100 mg/kg of rosemary extract for 12 weeks); (5) exercise and extract group (received 100 mg/kg of rosemary extract and concurrently put on a turned-on treadmill for 12 weeks). After the treatment, the testis tissue of rats was collected, and the seminiferous tubular diameter, luminal diameter, and epithelial height were analyzed. Also, spermatogenic lineage was counted in different groups. Results: The tubal diameter and epithelial height significantly increased following the consumption of rosemary extract and exercise training compared with the control group (P < 0.01). Also, a significant increase was observed in the luminal diameter in the exercise group compared to the control group (P < 0.01). The rosemary extract and exercise training significantly increased the number of spermatogonia cells (P < 0.01). However, there were no significant differences in the number of primary spermatocyte and spermatozoa cells among the different groups (P > 0.05). Conclusions: Generally, low-intensity aerobic exercise improved testicular histomorphology parameters. However, rosemary extract had no positive effects as an aerobic exercise on male fertility during aging.

Suppression of fertility by <em>Capparis spinosa</em> root extract through antioxidant alteration in reproductive system of male rats

Aim: To evaluate the suppression of fertility by oral mode of Capparis spinosa root extract through alteration in antioxidant defence of the reproductive system of male albino rats. Methods: The methanol extract of Capparis spinosa root (100mg/kg/day) was fed for 60 days to male, Wistar strain, albino rats. The fertility and testicular functions were assessed by mating tests, antioxidant defence indices, and spermatogenesis, histometry, and biochemical estimations for fertility suppression along with serum biochemistry and hematology for assessment of toxicology. Results: Capparis spinosa root extract administration for a period of 60 days did not cause body weight loss significantly, while the weight of testes, Cauda epididymis, seminal vesicle, and ventral prostate were significantly reduced. The lipid peroxidation and antioxidants levels i.e. superoxide dismutase, catalase, glutathione reduced, glutathione peroxidase, vitamin C, and Vitamin E in testes, Cauda epididymis, seminal vesicle, and ventral prostate were altered significantly (P<0.001) after the 60 days treatment of Capparis spinosa extract when compared to the controls. The root extract feeding caused a marked reduction in the number of primary spermatocytes, secondary spermatocytes, and spermatids however spermatogonia population showed non-significant change. Sertoli cell population was also affected. The number of mature Leydig cells was decreased and degenerating cells increased proportionately. The seminiferous tubule and Leydig cell nuclear area were reduced significantly (P<0.001) when compared to the controls. The protein and sialic acid content of the testes, Cauda epididymis, seminal vesicle, and ventral prostate were significantly (P<0.001) decreased. The glycogen in testes and fructose in seminal vesicle was also significantly reduced, whereas the testicular cholesterol was elevated as compared with the controls. Reduced sperm count and motility resulted in a total suppression of fertility. The RBC and WBC count, hemoglobin, and hematocrit in whole blood whereas marker enzymes and glucose in serum were found to be within the normal range after the 60 days of Capparis spinosa extract treatment. Conclusion: Capparis spinosa root extract may selectively act on developing germ cells mediated via Sertoli cells through increased oxidative stress or impairment of antioxidant defence system in reproductive organs of male rats, leading to suppression of fertility and suggests the feasibility of developing non-toxic, herbal male contraceptive drug.

Effects of MT on growth and gonad development of natural overwintering Whitmania pigra

The objectives of study were to explore the effects of exogenous methyltestosterone( MT) on the growth and gonadal development of overwintering Whitmania pigra. Before overwintering,0. 1,1. 0,10. 0,100. 0,150. 0 μg·L-1 of MT were added to the aquaculture water for 6 weeks. The changes of growth performance,gonad index,endogenous steroid hormones level and internal quality were measured after hibernate for 60 days. Then the tissue slice technique was used to observe the spermary( ovary) of Wh. pigra.The results showed that the body weight,survival rate and gonadal index increased first and then decreased with the increase of exogenous MT concentration; the male gland index was found the highest at the concentration of MT 10. 0 μg·L-1 and the female gland index was the highest at the concentration of MT 1. 0 μg·L-1. The survival rate of Wh. pigra peaked at the concentration of MT 10. 0 μg·L-1.The weight reaches a peak at a concentration of MT 100. 0 μg·L-1( P<0. 05). The number of primary spermatocytes in the testis was negatively correlated with the concentration of exogenous MT. The number of secondary spermatocytes and sperm cells increased first and then decreased. The concentration of secondary spermatocytes was the highest when the concentration of MT was 100. 0 μg·L-1.The number and volume of oocytes in the ovary and the yolk granules increased first and then decreased with the increase of exogenous MT concentration,and the highest was observed at the MT concentration of 100. 0 μg·L-1. The endogenous steroid hormone of Wh.pigra increased first and then decreased with the increase of exogenous MT concentration. The concentration of androgen and progesterone was the highest in MT 100. 0 μg·L-1 treatment( P<0. 05),and the concentration of estrogen was found the highest in MT 10. 0 μg·L-1 treatment( P<0. 05). After adding exogenous MT,Wh. pigra moisture content,acid-insoluble ash content,p H and anti-thrombin activity met the quality criterion of medicinal Wh. pigra in Chinese Pharmacopoeia( 2015 edition). In conclusion,the short-term addition of 1. 0-100. 0 μg·L-1 exogenous MT before hibernation can promote the growth,the development of sperm cells and the antithrombin activity of Wh. pigra.

The Effects of Olive Leaf Extract on The Testis, Sperm Quality and Testicular Germ Cell Apoptosis in Male Rats Exposed to Busulfan.

BackgroundBusulfan (BU) has a destructive effect on the male reproductive system. The goal of this study was to assess the effects of olive leaf extract (OLE) as a source of antioxidants and phenolic compounds, on BU-induced damages in rat testes.Materials and MethodsIn this experimental study, 40 male Wistar rats were randomly divided into 5 groups. The control group (CTL) received a single intraperitoneal (i.p.) injection of dimethyl sulfoxide (DMSO), followed by oral administration of distilled water for 5 weeks. In BU group, BU (10 mg/kg) was administrated i.p. once. In co- treatment groups, first, received BU (10 mg/kg, a single i.p. injection) then, OLE was administrated orally at different doses of 250 mg/kg (BU+OLE 250), 500 mg/kg (BU+OLE 500) and 750 mg/kg (BU+OLE 750), for 5 weeks. Next, blood and sperm samples were collected. The left testis was removed to investigate testicular parameters and apop- tosis by using H&E and TUNEL staining, respectively. All data were analyzed by SPSS software and a P<0.05 was considered significant.ResultsThere was a significant decline in sperm viability (P=0.017), number of primary spermatocyte (PS) (P=0.001) and Leydig cells (P=0.023) in the BU group versus the CTL group. OLE at three doses could repair these defects ver- sus BU group. Increases in apoptotic spermatogonia cells (SG) due to BU were significantly reduced by OLE 250 and 500 mg/kg (P<0.01). A reduction in germinal epithelium height and an increase in apoptotic SG were observed in BU+OLE 750 group vs. other groups (P<0.01) and alkaline phosphatase (ALP) was at the highest level, also Aspartate aminotransferase (AST) increased markedly vs. CTL (P=0.024).ConclusionOral administration of OLE at the doses of 250 and 500 mg/kg could be helpful in ameliorating BU- induced toxicity in rat testes, while OLE 750 mg/kg not only did not cause positive effects, but also could exacerbate the harmful effects.

Free radical scavenging potential of Drosera indica L in presence of Dalton Ascites lymphoma (DAL) tumor bearing mice

Exploration to find out new natural contraceptive agent for male is still being developed. Black pepper ( Piper nigrum L.) and its main alkaloid piperine have potential antifertility because of sitotoxic and hormonal effects. The aim of this study is to find out the effect of ethanolic extract of black pepper ( Piper nigrum L.) fruit of on reproductive hormone serum level, sperm quality, and spermatogenic cell populations in adult male Wistar rat. Twenty five male rats were divided into five groups consisting of two control group, i.e. K (-) (Na-CMC 0.5%), K (+) (finasteride 0.45mg/kg BW), and three groups received different doses of black pepper fruit ethanolic extract, i.e. D (1) (3.33mg/kg BW), D (2) (6.66mg/kg BW) and D (3) (13.32mg/kg BW) respectively. The treatment were given to each group for 55 days. Reproductive parameters were measured, including serum level of reproductive hormones (FSH, LH, dan testosteron), quality of cauda epididymal sperm (spermatozoa concentration, motility and morphological abnormality), spermatogenic cell populations (primary spermato-cyte and spermatid count) and seminiferous tubules diameter. Ethanol extract of black pepper fruit at doses of 3.33mg/kg BW, 6.66mg/kg BW, and 13.32mg/kg BW increased serum FSH level. Extract at dose of 13.32mg/kg BW decreased serum LH level, while extract at doses of 6.66mg/kg BW and 13.32mg/kg BW decreased serum testosterone level. The number of primary spermatocytes, spermatozoa concentration, and spermatozoa motility were decreased by administration of ethanol extract of black pepper fruit with dose of 6.66mg/kg BW and 13.32mg/kg BW. Ethanol extract of black pepper fruit at dose of 6.66mg/kg BW and 13.32mg/kg BW had a negative impact on the male reproductive system and showing potential antifertility in male rat.

The structure of spermatogenic cysts and number of Sertoli cells in the testes of Bombina bombina and Bombina variegata (Bombinatoridae, Anura, Amphibia)

Spermatogenesis in amphibians occurs inside spermatogenic cysts formed by Sertoli cells (SCs). SCs do not constitute a permanent population and their number increases during spermatogenic cycle and ontogeny. The number of SCs per cyst is strongly correlated with the number of germ cells in the same cyst and with the cyst volume. The number of germ and Sertoli cells and cyst volumes are species-specific and increase along with age. The mean number of primary spermatocytes per one Sertoli cell (efficiency) was the same, irrespective of age and species. Late spermatids are not attached to the Sertoli cells and do not form bundles as in other amphibians. The number of primary spermatocytes inside a cyst was the yield of several mitotic cycles of a single primary spermatogonium (stem cell) and its descendants, i.e., secondary spermatogonia. In adult Bombina bombina, the majority of cysts contained germ cells after 6–8 cycles, in juveniles after 4–6 cycles, and in adult Bombina variegata after 5–7 cycles. The mean numbers of SCs per cyst were 27.4 ± 9.9 in adult B. bombina, 6.42 ± 2.32 in adult B. variegata, and 7.1 ± 2.33 in juvenile B. bombina. The number of primary spermatocytes per one SC (SC efficiency) in B. bombina in adults was 6.5 ± 2.2, in juveniles 9.0 ± 4.2, and in adult B. variegata 7.40 ± 2.96. Spermatogenesis in Bombinatoridae is exceptional owing to the lack of intimate contact between SCs and germ cells.

Histological and statistical studies on the gonadal cycle of the mussel Lithophaga truncata (Gray, 1843) along immature and mature phases (Pelecypoda - Mytilidae)

The main purpose of this study was to investigate the gonadal cycle of the musselLithophaga truncata (Gray, 1843). Specimens were sampled regularly twice every season at depth 1- 2 meters in the year (2015 - 2016) along the northern estuarine harbor of the Arabian Gulf - Saudi Arabia. Parts of gonads (ovaries and testes) were prepared for histological and statistical studies. The histological observations of the gonad indicated that there was only one gonadal cycle in the year. The ovaries produced oocytes which undergone vitellogenesis to change these oocytes into comparatively ripe ones condensed with yolk. With slide micrometer a fixed length and width of the ovary was determined in serial histological sections (30000 µm). The different stages of oocyte development were counted and statistically analyzed. During November - February the gonadal follicles appeared small in size and in a quiescent state. Follicles were few in number and scattered throughout the vesicular connective tissue. In males the follicles were small and scattered throughout the connective tissue in the same way as females. Follicles contained undifferentiated residual cells, spermatogonia and negligible number of primary spermatocytes. The oogonia were the most dominant type. Few number of previtellogenic oocytes were observed. The ooplasm of these stages, oogonia and previtellogenic oocytes, was homogeneous and with comparatively large nucleus. In previtellogenic oocytes small granules of the same nature were scattered randomly in the ooplasm. These granules consisted of proteins (peptide linkage). Carbohydrates and lipids were entirely absent.During April - July previtellogenic oocytes began to undergo growth phase and extending towards the center of the follicles. In males spermatogenesis became more apparent and proceeded at a faster rate. In some mussels spermiogenesis was formed in great number. The gonadal follicles ramified rapidly, and the development of gametes of both sexes was very fast. In females the vitellogenic oocytes showed prominent development. Atretic oocytes and their debris were negligible. Very few number of oogonia were observed. Yolk bodies (glycoproteins) increased rapidly and replaced the vacuoles in the ooplasm. Other large granules gained a dark blue color when Aqueuos bromophenol blue was applied. These granules are basic proteins. Other large-sized granules began to appear at the periphery of the ooplasm. These granules gained positive reaction with Alcian blue, PAS and Alcian blue- PAS reactions. The vitelline envelope became conspicuous under the follicle cells. Mucosubstances were positively intense at the periphery of the oocyte while proteins surrounded the nucleus. Gradually fat deposition increased. The size range of these oocytes was 130 - 180µm with the ratio of the nucleus to the ooplasm 0.5:2. During August - October both sexes had ripe gametes. In males, the central portion of the follicles was empty because of the recent discharge of large quantities of ripe gametes. At the end of this step resorption of the gonads proceeded very rapidly and emptied follicles began to acquire an elongated form. At this step the oocyte attains its maximal size 180 - 200µm. The mean of the total number of oocyte developmental stages in all seasons showed that the ovary was active in all seasons of the year but different labors were present. It concluded that the oogonia , the postvitellogenic oocyte development and atretic oocytes were conspicuous during the first step of oogenesis ; the vitellogenic oocytes were dominant during the second step of oogenesis whereas the postvitellogenic and atretic oocytes were dominant during the third step of oogenesis. All data obtained were subjected to analysis of variance (ANOVA) means significant difference P>0.05 or P 0.05.

Anti-fertility effect of hydro-alcoholic extract of fennel (Foeniculum vulgare Mill) seed in male Wistar rats

Abstract Introduction: The study aimed to investigate the anti-fertility effect of fennel (Foeniculim vulgare Mill) seed extract in male rats. Material and Methods: Forty Wistar rats were divided into five equal groups. The control group received distilled water and the experimental groups were orally administered 1 ml of hydro-alcoholic extract of fennel seed in four doses of 35, 70, 140, and 280 mg/kg/b.w. daily for 60 days. After the last gavage, the rats were anaesthetised and the caudal part of the right epididymis was used for sperm counting. After fixation of the testes, microscopic sections were prepared and histological changes were evaluated. Results: The number of spermatogonia after doses of 140 and 280 mg/kg and Sertoli cells after a dose of 140 mg/kg decreased significantly as compared with the control group (P &lt; 0.05). The number of primary spermatocytes and sperm count decreased significantly in the experimental groups (70, 140, and 280 mg/kg) when compared to the control group (P &lt; 0.05). Furthermore, thickening of the basement membrane, cell apoptosis, and irregular arrangement of the germinal epithelium were observed in the experimental groups. Conclusion: Hydro-alcoholic fennel seed extract at these doses could reduce reproductivity and has anti-fertility activity in male rats.

Testicular cell indices and peripheral blood testosterone concentrations in relation to age and semen quality in crossbred (holstein friesian×tharparkar) bulls.

Present study analyzed the changes in peripheral blood testosterone concentrations and testicular cytogram in relation to age and semen quality in crossbred males. Three different age groups of crossbred males viz. bull calves (6 months, n = 5), young bulls (15 months, n = 5) and adult bulls (4 to 6 years, n = 8) were utilized for the study. Testicular fine needle aspiration cytology technique was used to quantify testicular cytology and their indices. Peripheral blood testosterone concentrations were measured using enzyme-linked immunosorbent assay method. Semen samples collected from adult bulls were microscopically evaluated for quality parameters. Mean peripheral blood testosterone concentrations in bull calves, young bulls and adult bulls were 2.28±0.09 ng/mL, 1.42±0.22 ng/mL and 5.66±1.08 ng/mL respectively, and that in adult bulls were significantly different (p<0.01) from young bulls and bull calves. There was no significant difference between the proportion of different testicular cells in bull calves and young bulls. Between young and adult bulls, significant differences (p<0.01) were observed in the proportion of spermatocytes, spermatozoa, and sperm: Sertoli cell ratio. The proportions of Sertoli cells showed a significant difference (p<0.01) between the three age groups. The number of primary spermatocytes had a positive correlation with peripheral blood testosterone concentrations in bull calves (r = 0.719, p<0.01). Number of Sertoli cells per 100 germ cells was negatively correlated with blood testosterone concentration in young bulls (r = −0.713, p<0.01). Among different semen parameters in adult bulls, ejaculate volume (r = 0.790, p<0.05) had positive relationship, and sperm motility had significant negative correlation (r = −0.711, p<0.05) with testosterone concentrations. The number of Sertoli cells and Sertoli cell index had a positive correlation with various semen quality parameters (p<0.001). Results of the present study conclude that number of Sertoli cells and Sertoli cell index are good indicators of semen quality, but peripheral blood testosterone concentrations may not have a direct relationship with various seminal attributes in crossbred bulls.

The Effects of Hydroalcoholic Extract of Apium graveolens Leaf on the Number of Sexual Cells and Testicular Structure in Rat.

Use of medicinal plants with high antioxidant properties could be effective to increase fertility and improvement of disorders such as hormonal imbalance, impotency, oligospermia and immotile sperm. Celery (Apium graveolens) is rich in antioxidant agents. The leaf and stems of celery contain phenols, furanocoumarin and luteolin. Apigenin is one of the main flavonoids of celery leaf. This study aimed to investigate the effects of hydroalcoholic extract of celery on histological properties of testis and number of sexual cells in male rats. Thirty-two male Wistar rats were divided into four groups of eight rats each. Control, did not receive any medication; sham, received normal saline; and two groups received celery extract orally in dosages of 100 and 200 mg/kg/BW once every two days for 60 days. At the end, animals were anesthetized, and caudal part of the right epididymis was used for sperm counting. After fixation of testis, tissue sections were prepared and studied microscopically to evaluate morphometric (lumen diameter, number of primary spermatocyte and sertoli cell) and histological changes. Data was analyzed by one-way ANOVA test using SPSS15 software. P < 0.05 was considered as statistically significant. There was a significant increase in the number of sperms, sertoli cells, and primary spermatocyte (P < 0.05) in groups receiving extract; however, structural changes were not observed in the groups. It seems that celery increases spermatogenesis in male rats, but has no destructive effects on testicular tissue.

Effects of opioid (tramadol) treatment on testicular functions in adult male rats: The role of nitric oxide and oxidative stress

Nowadays, tramadol hydrochloride is frequently used as a pain reliever, and for the treatment of premature ejaculation. Decreased semen quality was noted in chronic tramadol users. The present study aimed to elucidate the effects of tramadol on the testicular functions of adult male rats. A total of 40 albino adult male rats were divided into control and tramadol groups, with 20 rats for each group. Rats of the tramadol group were subcutaneously injected with 40 mg/kg three times per week for 8 weeks. The control group received normal saline 0.9%. Blood samples from each animal were obtained. Plasma levels of different biochemical substances were determined. Nitric oxide was measured in testicular tissue samples. Those samples together with epididymal tissue samples were processed for histopathological examination. Tramadol significantly reduced plasma levels of luteinizing hormone, follicle-stimulating hormone, testosterone and total cholesterol, but elevated prolactin and estradiol levels compared with the control group. In addition, tramadol increased the testicular levels of nitric oxide and lipid peroxidation, and decreased the anti-oxidant enzymes activities significantly compared with the control group. The tramadol group showed decreased sperm count and motility, and numbers of primary spermatocytes, rounded spermatid and Leydig cells. Immunohistochemical examinations showed that tramadol increased the expression of endothelial nitric oxide synthase in testicular tissues. The present study showed that tramadol treatment affects the testicular function of adult male rats, and these effects might be through the overproduction of nitric oxide and oxidative stress induced by this drug.

Down-regulation of metallothionein 1 and 2 after exposure to electromagnetic field in mouse testis.

It is proved that testis is sensitive to electromagnetic field (EMF) and its damage results in infertility. Exposure to EMF induces reactive oxygen species production and affects on anti-oxidants defense mechanisms. Metallothionein (MT) is a name for a group of low molecular weight (6-7 kDa), sulfhydryl rich proteins. Expression of MT1 and MT2 genes in testis tissue after EMF exposure was aimed in this study. Male BALB/c mice (8 weeks old) were exposed to 3 MT EMF for 8 weeks, 4 hours/day. After 8 weeks, the mice were sacrificed and the testis tissue was removed. The testis pieces were stained with hematoxylin and eosin and analyzed under an optical microscope. Assessment of MT1 and MT2 genes and also protein expression was performed by real-time PCR and Western-blot, respectively. In light microscopic observation, the number of primary spermatocytes was increased significantly in EMF group (P < 0.01). In addition, in interstitial space, the number of leydig cells was increased significantly in EMF group (P < 0.01) and basement membrane thickness was increased as well. MT1 and MT2 genes were down-regulated significantly in testis tissue of mice exposed to EMF both in mRNA and protein level compared to control. It is clear that MT is mediated in testis development and spermatogenesis. Down-regulation of MT1 and MT2 after EMF in mouse testis might be followed by some consequences that result in infertility.

The toxic effects of sodium fluoride on the reproductive system of male rats

The present study was undertaken to evaluate the effect of fluoride toxicity on the reproductive system of male rats. Sexually mature male Wistar rats were exposed to 2, 4, and 6 ppm sodium fluoride in their drinking water for 6 months ad libitum. Sperm motility and density in cauda epididymis were assessed. Biochemical and histological analysis were performed in reproductive organs. Fluoride treatment brought about a significant decrease in the weight of testis, epididymis, and ventral prostate. The sperm motility and density were significantly reduced. There was a marked reduction in the number of primary spermatocyte, secondary spermatocyte, and spermatids. The Sertoli cell counts and their cross sectional surface areas were significantly decreased. The Leydig cell nuclear area and the number of mature Leydig cells were also significantly decreased. The protein content of the testis and epididymis were significantly reduced. Fructose in the seminal vesicles and cholesterol in testes were increased significantly. In conclusion, sodium fluoride administrated in drinking water of 2, 4, and 6 ppm concentration for 6 months to male rats adversely affected their fertility and reproductive system.

Effects of hormonal treatment on the contralateral descended testis in unilateral cryptorchidism

Objective One of the therapeutic options in the treatment of cryptorchidism is hormonal therapy with luteinizing hormone releasing hormone (LHRH) and/or human chorionic gonadotropin (HCG); concerns have, however, been raised regarding its safety. The purpose of this study was to demonstrate that hormonal therapy improves the abnormal histology of the contralateral descended testis without harming the germ cells. Method Patients with unilateral cryptorchidism were randomized into two groups: those treated with orchiopexy alone and those treated with LHRH long acting analog and HCG. Biopsies taken from contralateral descended testes were analyzed and compared with controls. Results The number of germ cells per tubule in contralateral testes of patients treated with orchiopexy alone is significantly lower than the number of germ cells in testes of patients with spontaneously descended testes ( P < 0.0001). Hormonal therapy did not have any adverse effect upon the histology of the contralateral testis, but in fact improved it. Seven weeks of hormonal therapy induced a rise in the number of germ cells per tubule ( P < 0.05). It was also beneficial for the number of adult dark spermatogonia per tubule and the number of primary spermatocytes, although these differences did not reach statistical significance. Conclusion The contralateral testis in patients with unilateral cryptorchidism is abnormal. Hormonal therapy improves the histopathology of the contralateral testis without harming the germ cells.

Effect of Cressa cretica. Methanol Extract on Testicular Function of Albino Rats

Oral administration of a methanolic extract of Cressa cretica. Linn. (Rundati) (Convolvulaceae) (whole plant) at a dose level of 100 mg/kg/day for a period of 60 days led to a significant decrease in the weight of testis, epididymis, seminal vesicle, and ventral prostate. Cressa cretica. reduced the fertility of male rats by 100%. There was a marked reduction in the number of primary spermatocytes, secondary spermatocyte, and spermatids. Sertoli cell counts as well as the cross-sectional surface area were significantly decreased. Leydig cell nuclear area and the number of mature Leydig cells were also significantly decreased. The protein, sialic acid, glycogen, and cholesterol content of the testis, the fructose in the seminal vesicle, and protein and sialic acid in the epididymis were significantly decreased. Serum testosterone levels were also reduced after Cressa cretica. treatment. The RBC and WBC counts, hemoglobin, hematocrit, blood sugar, serum cholesterol, phospholipids, triglyceride, and HDL-cholesterol were within the normal range. The methanol extract of Cressa cretica. produced intrusion in testosterone production and affected spermatogenesis in male albino rats.

Effects of acid sphingomyelinase deficiency on male germ cell development and programmed cell death.

Deficiency of acid sphingomyelinase (ASM), an enzyme responsible for producing a pro-apoptotic second messenger ceramide, has previously been shown to promote the survival of fetal mouse oocytes in vivo and to protect oocytes from chemotherapy-induced apoptosis in vitro. Here we investigated the effects of ASM deficiency on testicular germ cell development and on the ability of germ cells to undergo apoptosis. At the age of 20 weeks, ASM knock-out (ASMKO) sperm concentrations were comparable with wild-type (WT) sperm concentrations, whereas sperm motility was seriously affected. ASMKO testes contained significantly elevated levels of sphingomyelin at the age of 8 weeks as detected by high-performance, thin-layer chromatography. Electron microscopy revealed that the testes started to accumulate pathological vesicles in Sertoli cells and in the interstitium at the age of 21 days. Irradiation of WT and ASMKO mice did not elevate intratesticular ceramide levels at 16 h after irradiation. In situ end labeling of apoptotic cells also showed a similar degree of cell death in both groups. After a 21-day recovery period, the numbers of primary spermatocytes and spermatogonia at G2 as well as spermatids were essentially the same in the WT and ASMKO testes, as detected by flow cytometry. In serum-free cultures both ASMKO and WT germ cells showed a significant increase in the level of ceramide, as well as massive apoptosis. In conclusion, ASM is required for maintenance of normal sphingomyelin levels in the testis and for normal sperm motility, but not for testicular ceramide production or for the ability of the germ cells to undergo apoptosis.

Protection from radiation-induced male germ cell loss by sphingosine-1-phosphate.

Male germ cells are susceptible to radiation-induced injury, and infertility is a common problem after total-body irradiation. Here we investigated, first, the effects of irradiation on germ cells in mouse testis and, second, the role of sphingosine-1-phosphate (S1P) treatment in radiation-induced male germ cell loss. Irradiation of mouse testes mainly damaged the early developmental stages of spermatogonia. The damage was seen by means of DNA flow cytometry 21 days after irradiation as decreasing numbers of spermatocytes and spermatids with increasing amounts of ionizing radiation (0.1-2.0 Gy). Intratesticular injections of S1P given 1-2 h before irradiation (0.5 Gy) did not protect against short-term germ cell loss as measured by in situ end labeling of DNA fragmentation 16 h after irradiation. However, after 21 days, in the S1P-treated testes, the numbers of primary spermatocytes and spermatogonia at G2 (4C peak as measured by flow cytometry) were higher at all stages of spermatogenesis compared with vehicle-treated testes, indicating protection of early spermatogonia by S1P, whereas the spermatid (1C) populations were similar. In conclusion, S1P appears to protect partially (16%-47%) testicular germ cells against radiation-induced cell death. This warrants further studies aimed at development of therapeutic agents capable of blocking sphingomyelin-induced pathways of germ cell loss.

Seasonal changes in the seminiferous epithelium of rhesus and bonnet monkeys.

With a view to elucidate seasonal variations in testicular spermatogenesis, quantitative analysis of spermatogenic cells was carried out in non-human primate species viz. rhesus (Macaca mulatta) and bonnet (M. radiata) monkeys during breeding (October-December) and non-breeding (May-June) seasons. The results revealed significant inhibition of testicular germ cell population during non-breeding compared with the breeding period in both the species. Quantitative determination of Sertoli cell-germ cell ratio showed a marked decrease in the number of type A-spermatogonia, spermatocytes (non-pachytene and pachytene) and spermatids (in steps 1-12 of spermiogenesis) in rhesus monkey during the non-breeding period. Bonnet monkeys exhibited the significant decline in the number of primary spermatocytes and spermatids during the non-breeding phase. In addition, average diameter of round seminiferous tubules and nuclear diameter of Leydig cells also decreased significantly in rhesus monkeys. However, bonnet monkeys did not show any significant change in nuclear diameter/morphology of Leydig cells, testicular tubular diameter and number of type A-spermatogoniae. Sertoli cell number did not show any significant change during both breeding and non-breeding periods in both the species. The results of this study indicate a prominent seasonal variation in testicular spermatogenic/Leydig cells in rhesus monkeys than those observed in bonnet monkeys.

Dynamics of testicular germ cell proliferation in normal mice and transgenic mice overexpressing rat androgen-binding protein: a flow cytometric evaluation.

Transgenic mice carrying rat androgen-binding protein (ABP) genomic DNA express high amounts of testicular ABP and develop a progressive impairment of spermatogenesis. To understand the mechanism of these changes, we have studied the pattern of testicular germ cell proliferation from 7 to 360 days of age in wild-type (WT) control and transgenic homozygous (ABP-TG) mice by flow cytometry after labeling DNA in isolated germ cells with propidium iodide. At all ages studied, the body weight of the ABP-TG mice was lower than that of age-matched WT controls. Significantly reduced testicular weight and total germ cell number in the ABP-TG mice were evident from Day 30 and Day 60, respectively. Flow cytometric analysis of isolated germ cells revealed that the number of germ cells undergoing proliferation (S-phase cells) was identical in WT control and ABP-TG mice up to Day 14. Subsequently, the number of germ cells in S-phase was consistently higher in ABP-TG than in WT mice. The number of primary spermatocytes was significantly increased starting from Day 60, and the numbers of round and elongated spermatids were significantly reduced in the ABP-TG animals from Day 21 and Day 60 onwards, respectively. Immunocytometry for intracellular ABP at 90 days of age revealed that the percentage of ABP-containing germ cells was greater in ABP-TG than in WT mice. The continuous presence of ABP in mouse seminiferous tubules at greater than physiological concentrations facilitates the formation of primary spermatocytes but impairs subsequent transformation to round and elongated spermatids. Based on our observations and the analysis of the available literature, the most likely mechanism for production of these effects is sustained reduction in the bioavailability of androgens.