Prepubertal Rat Testis Research Articles

Possible protective effect of Crocin on Cisplatin-induced changes in the testis of prepubertal albino rats: Histological, Immunohistochemical, and Morphometric Study

Possible protective effect of Crocin on Cisplatin-induced changes in the testis of prepubertal albino rats: Histological, Immunohistochemical, and Morphometric Study

Histomorphological and histomorphometric changes in the testes of male Wistar rats following prepubertal exposure to caffeine

This experiment was designed to study the effects of caffeine on the histomorphology of the testes of prepubertal male Wistar rats. Prepubertal Wistar rats (25-28 days old), weighing between 50-60 g were randomly divided into three treatment groups (B, C and D) and control (A) of ten rats each. Group A rats received distilled water while Groups B, C, and D rats were respectively given daily dose of 50 mg/kg, 100 mg/kg, and 200 mg/kg of caffeine dissolved in distilled water for 14 days by gavage. Five rats from each group were sacrificed 24 hours after the last caffeine administration while the remaining rats were monitored for another three weeks for them to attain sexual maturity. The rats were sacrificed under diethyl ether anesthesia and the testes were fixed by immersion in Bouin’s fluid. The tissues were processed for histomorphological and histomorphometric assessment. Data obtained were analyzed using one way ANOVA followed by Dunnett post hoc test for comparison with control. Alpha level was set at 0.05. Results showed that treatment with caffeine caused a significant decrease in testicular weight and a marked cellular degeneration and depletion of germ cells in the testes of rats in group D. The height of the germinal epithelium was significantly reduced when compared with control. This study concluded that prepubertal exposure of Wistar rats to caffeine reduced testicular growth and deleteriously alters testicular morphology.

LOCAL-IGF-1 and GH application IMPROVES germ cell histology, spermatogenesis and fertility after experimental testicular torsion and detorsion

To evaluate the impact of insulin like growth factor-1(IGF-1) and growth hormone (GH) on testis histology, spermatogenesis, and fertility in prepubertal rats exposed to 6h of testicular torsion (TT) and detorsion. Forty-eight male Wistar-albino rats weighing 30-70g and at 3-week age were allocated into six groups involving eight rats in each group as follows: Group 1:Sham, Group 2:Control, Group 3:Gelatin, Group 4:Local-IGF-1, 5: Local-GH, Group 6: Systemic-GH. Right testis was only exposed and sutured in the sham group, and right testes were rotated clockwise, 720°, fixed, and 6h later, detorsion on the testis was done in groups 2-6. Unloaded gelatin, 5μg local-IGF-1 loaded, and 2IU rhGH loaded gelatin were sutured to the right testis after detorsion in groups 3-5. In Group 6, 0.3IU/100gr/d rhGH was given for seven days via subcuticular route after detorsion. Each of the rats cohabited with two female rats five weeks later. Afterward, both right and left testes were removed. Mean diameter of seminiferous tubules (STD), mean biopsy score count of the testis (TBSC), mean percentage of haploid cells (HCP) were assessed, and fertility parameters were evaluated. STD and TBSC of the ipsilateral testes were significantly reduced in control and gelatin groups when compared to sham, local-IGF-1, and local-GH groups. STD and TBSC of the ipsilateral testes of the systemic-GH group were decreased compared to the sham group. HCP of the ipsilateral testes of control, gelatin, and systemic-GH groups were significantly lower than the sham, local-IGF-1, and local-GH groups. STD, TBSC, and HCP of the contralateral testes were significantly reduced in control and gelatin groups when compared separately to sham, local-IGF-1, systemic- GH, and local-GH groups. The difference between groups regarding potency, fertility, fecundity indexes, and mean fetus numbers were not significant. Even though there was significant and permanent histologic germ cell damage and reduced HCP in both ipsilateral and contralateral testes, experimental 6h TT and detorsion in prepubertal rats did not have a negative impact on future fertility. Local-IGF-1and rhGH treatment improved germ cell histology and spermatogenesis in both ipsilateral and contralateral testes of prepubertal rats, subjected to 6h of TT and detorsion.

Delayed Consequences of the Toxic Effect of Paclitaxel on the Testes of Prepubertal Rats and Their Correction with p-Tyrosol.

Delayed gonadotoxic effects were revealed in outbred male sexually mature rats (SD) after exposure to paclitaxel in the prepubertal period, and the possibility of their correction with p-tyrosol was shown. It was found, that administration of paclitaxel does not inhibit the ability of animals to conceive, but impairs the reserve capacity of the testicular tissue. In intact female rats crossed with male rats receiving paclitaxel, increased post-implantation fetal death was observed. Combined administration of paclitaxel and p-tyrosol alleviated the delayed effects of the cytostatic treatment on the prepubertal testis.

Oxidative Stress Disrupted Prepubertal Rat Testicular Development after Xenotransplantation.

In the past two decades, testicular tissue grafting and xenografting have been well established, with the production of fertilization-competent sperm in some studies. However, few studies have been carried out to observe the development of grafted prepubertal testicular tissue of rats and compare the biological differences between in situ testis and grafted testis. In this study, we established the prepubertal testicular tissue xenografting model using a 22-day-old rat and evaluated certain parameters, including testicular histology, testosterone production, and ultrastructure of the grafted testes. We also assessed gene expression of cell proliferation markers, testicular cell markers, and antioxidative defense system. Our results showed that 47 days after transplantation, intratesticular testosterone concentration was not significantly altered; however, cell proliferation, spermatogenesis, and Sertoli cell markers in the transplanted testes were significantly disrupted compared with the control group, accompanied by aggravated apoptosis and oxidative damage. Moreover, the transplanted testes showed smaller tubular diameter and disrupted spermatogenic epithelium with apparent vacuoles, distorted and degenerated germ cells with obscure nuclear margin, and no spermatids in the center of the tubules. Although testis xenografting has been extensively tested and attained great achievement in other species, the prepubertal rat testicular tissue xenografting to immunodeficient mice exhibited obvious spermatogenesis arrest and oxidative damage. The protocol still needs further optimization, and there are still some unknown factors in prepubertal rat testes transplantation.

Effect of Silver Nanoparticles on Testes of Prepubertal Male Albino Rats and the Possible Protective Role of Vitamin E Supplement (Histological and Immunohistochemical Study)

Background: Silver nanoparticles (AgNPs) are one of the most eminent and commonly used nanoparticles in customer everyday products for both adults and children.Objective: This study objects to study the effect of prepubertal exposure of AgNPs on the histological structure of testis and the possible protective role of vitamin E.Materials and Methods: Twenty three days old juvenile male albino rats were used and divided into 3 groups. Group I control: subdivided into two subgroups: subgroup Ia (negative control), Rats received no treatment and sacrified at postnatal day (PND) 23 and 58. Subgroup Ib (vehicle control group), further subdivided into two subgroup Ib1, Ib2. Group II: subdivided into 2 subgroups: Subgroup IIa; rats treated with AgNPs intraperitoneally daily at dose of (50 μg /kg/day) for 35 days from 23 PND to 58 PND and Subgroup IIb; treated with AgNPs and vitamin E (vit. E) orally by gavage at a dose (150 mg /kg /day) dissolved in corn oil from age 23-58 PND, Group III; follow up group, rats received AgNPs left without treatment till 90 PND.Results: Testicular damage was also detected in AgNPs treated group as compared to the control group. Vit. E-treated subgroup and the follow up groups showed significant testicular improvement. A drastic decrease of testicular germinal epithelial height in AgNPs treated compared to control and vit. E treated rats. The expression of Bcl2 immunoreaction was also decreased in AgNPs-treated rats compared to the control and AgNPs and vit. E-treated groups.Conclusion: AgNPs induced structural changes on male reproductive system that can be ameliorated by Vit. E supplementation and limiting the exposure to the products rich in these particles.

Troxerutin affects the male fertility in prepubertal type 1 diabetic male rats.

Objective(s):Diabetes can gradually cause damage to the function and structure of male gonads. This survey was conducted to investigate the effect of troxerutin on hormonal changes, serum oxidative stress indices, and testicular function and structure in prepubertal diabetic rats.Materials and Methods:Fifty prepubertal (6 weeks old) male Wistar rats were divided into five groups including Control, Troxerutin, Diabetic, Diabetic+Troxerutin, and Diabetic+Insulin. Type I diabetes was induced by 55 mg/kg of streptozotocin intraperitoneally. The groups were treated with 150 mg/kg/day troxerutin via oral gavage or 4-6 IU/day insulin via subcutaneous injection for 4 consecutive weeks. Blood sugar (BS) and serum levels of insulin, FSH, LH, testosterone, glutathione peroxidase (GPX), superoxide dismutase (SOD), malondialdehyde (MDA), and total antioxidant capacity (TAC) were analyzed. Testis and epididymis were removed for histopathologic study and analysis of sperm parameters.Results:Troxerutin significantly reduced the BS in the diabetic group similar to insulin but could not affect insulin, FSH, or LH significantly. Troxerutin caused a significant increase in testosterone and GPX but had no significant effect on serum MDA, TAC, and SOD levels. In addition, troxerutin had a better effect than insulin on diabetes-induced testicular structural damage. Sperm analysis results also revealed that troxerutin and insulin could improve sperm number, motility, and viability in diabetic rats.Conclusion:According to these results, it can be derived that administration of troxerutin is a suitable protective strategy for side effects of diabetes in testis of prepubertal diabetic male rats.

Perfluorododecanoic Acid Blocks Rat Leydig Cell Development during Prepuberty.

Perfluorododecanoic acid (PFDoA) has been used as a surfactant and may have reproductive toxicity. However, whether PFDoA influences Leydig cell development during prepuberty remains unknown. In the present study, 21-day-old male Sprague-Dawley rats were gavaged 0, 5, or 10 mg/kg PFDoA from postnatal day 21 to 35. PFDoA decreased the serum concentrations of testosterone, luteinizing hormone, and follicle-stimulating hormone at doses of 5 and 10 mg/kg without influencing Leydig cell number and proliferation. However, PFDoA down-regulated the expression of Leydig cell genes ( Lhcgr, Scarb1, Star, Cyp11a1, Cyp17a1, and Hsd11b1) or their proteins. PFDoA dose-dependently reduced SIRT1 and PGC-1α levels. PFDoA did not affect AMPK and AKT2 levels but decreased their phosphorylation. We also treated primary progenitor Leydig cells purified from prepubertal rat testes with PFDoA for 24 h. It in vitro lowered viability and decreased mitochondrial membrane potential of progenitor Leydig cells, but it stimulated the generation of the intracellular reactive oxygen species and induced Leydig cell apoptosis at 10 μM. In conclusion, PFDoA blocks rat Leydig cell development during the prepubertal period possibly via targeting AMPK/SIRT1/PGC-1α and AKT2 signaling pathways.

Mutual promotion of apoptosis and autophagy in prepubertal rat testes induced by joint exposure of bisphenol A and nonylphenol

BPA and NP are both typical endocrine disruptors, the exposed populations are widespread, and the health risks mustn't be ignored. However, the interactions between them on spermatogenesis are rarely mentioned. And the underlying mechanism is unclear yet. In the present study, prepubertal SD rats were exposed to different low doses of BPA and NP separately or jointly for 4 weeks. The results indicate that the joint exposure induced excessive apoptosis and autophagy in the testes, as proved by a series of characteristics such as chromatin condensation and autophagosomes formation. Besides, endocrine disorders and oxidative stress were also caused by the exposure. Apoptosis was mediated by the mitochondrial apoptosis pathway, since the Bax and Caspase-3 gene expressions significantly increased with a prominent decrease of Bcl-2. While autophagy was caused by the inhibition of the Akt/mTOR pathway, as the expressions of the downstream genes Beclin-1, Atg5, Atg12 and the split of LC3 protein increased altogether. Worse yet, autophagy and apoptosis might reinforce each other and make the situation more severe in the joint group. What's more, remarkable histopathological changes such as spermatogenic epithelium atrophy, germ cell loss, and various ultrastructural modifications were strongly related to the apoptosis and autophagy. In aggregate, this study shows the enormous risk on male reproductive system brought by the interactions between BPA and NP. The findings provide a broader vision to understand the roles of apoptosis and autophagy induced by the joint exposure in the aggravation of spermatogenesis impairment, which could be a reference for the situation of complex EDCs exposure-induced male reproductive toxicity, and possibly inspire us to find new ideas for preventive and therapeutic treatments.

EB 2017 Article: Soy protein isolate feeding does not result in reproductive toxicity in the pre-pubertal rat testis.

The isoflavone phytoestrogens found in the soy protein isolate used in soy infant formulas have been shown to have estrogenic actions in the developing male reproductive tract resulting in reproductive toxicity. However, few studies have examined potential estrogenicity of soy protein isolate as opposed to that of pure isoflavones. In this study, we fed weanling male Sprague-Dawley rats a semi-purified diet with casein or soy protein isolate as the sole protein source from postnatal day 21 to 33. Additional groups were fed casein or soy protein isolate and treated s.c. with 10 µg/kg/d estradiol via osmotic minipump. Estradiol treatment reduced testis, prostate weights, and serum androgen concentrations ( P < 0.05). Soy protein isolate had no effect. Estradiol up-regulated 489 and down-regulated 1237 testicular genes >1.5-fold ( P < 0.05). In contrast, soy protein isolate only significantly up-regulated expression of 162 genes and down-regulated 16 genes. The top 30 soy protein isolate-up-regulated genes shared 93% concordance with estradiol up-regulated genes. There was little overlap between soy protein isolate down-regulated genes and those down-regulated by estradiol treatment. Functional annotation analysis revealed significant differences in testicular biological processes affected by estradiol or soy protein isolate. Estradiol had major actions on genes involved in reproductive processes including down-regulation of testicular steroid synthesis and expression of steroid receptor activated receptor (Star) and cytochrome P450 17α-hydroxylase/(Cyp17a1). In contrast, soy protein isolate primarily affected pathways associated with macromolecule modifications including ubiquitination and histone methylation. Our results indicate that rather than acting as a weak estrogen in the developing testis, soy protein isolate appears to act as a selective estrogen receptor modulator with little effect on reproductive processes. Impact statement Soy protein isolate (SPI) is the sole protein used to make soy-based infant formulas. SPI contains phytoestrogens, which are structurally similar to estradiol. These phytoestrogens, daidzein, genistein, and equol, fit the definition of endocrine-disrupting compounds, and at high concentrations, have estrogenic actions resulting in reproductive toxicity in the developing male, when provided as isolated chemicals. However, few animal studies have examined the potential estrogenicity of SPI as opposed to pure isoflavones. In this study, SPI feeding did not elicit an estrogenic response in the testis nor any adverse outcomes including reduced testicular growth, or androgen production during early development in rats when compared to those receiving estradiol. These findings are consistent with emerging data showing no differences in reproductive development in males and female children that received breast milk, cow's milk formula, or soy infant formula during the postnatal feeding period.

Comperative Investigation of Structure of Male Genital System of Ductus Deferens Ligation in Rats

Objective: We investigated structural and morphological changes in pre-pubertal and pubertal rat testis and ductus deferens after unilateral vasectomy. Vasectomy is reliable and popular male contraception method, applied widely in the world. Methods: Six pre-pubertal and 6 pubertal male Wistar albino rats were divided into four groups. Unilateral ductus deferens ligation was performed on right ductus deferens of each rat. Right testis and ductus deferens tissues were evaluated as ligation group while left tissues were control groups. Two months after operation, the vas deferens and testes removed to obtain paraffin block sections and morphometric studies carried out with light microscopy. Hematoxylin-eosin staining and vascular endothelial growth factor (VEGF) immunohistochemically staining were applied. Results: Structural and morphological changes and decrease in spermatogenesis process have been observed at testis and ductus deferens tissues of all vasectomized rat. The seminiferous epithelium and the seminiferous tubule diameter were thickened. Degenerative changes have been observed at the epithelial structure of Ductus deferens and among muscle fibers forming the muscle of the Ductus deferens. Moreover, in pre-pubertal and pubertal vasectomized rat VEGF immunoreactivity was not observed in seminiferous tubule. Conclusion: The unilateral ligation operation can cause damage at ductus deferens and testis tissue in the pre-pubertal and pubertal period.

Immediate and late effects of chronic stress in the testes of prepubertal and adult rats.

The objective of this study was to investigate the effects of chronic stress on the testes of prepubertal and adult rats and to evaluate whether any alterations could be reversed when stress induction is ended. Seventy-six male rats were assigned to eight groups depending on the type of treatment (control or stressed), the age at which stress was initiated (prepubertal or adult), and the time of evaluation (immediate or late). Stress stimuli were applied for 6 weeks. Stressed prepubertal and adult rats evaluated immediately after the last stress stimulus were included in SP-I and SA-I groups, respectively. The late prepubertal (SP-L) and adult (SA-L) groups of stressed rats were evaluated 6 weeks after the last stress stimulus. Age-matched rats were used as controls (CP-I, CA-I, CP-L, and CA-L groups). Application of stress stimuli to rats in the SP-I group resulted in body weight and seminiferous tubule diameter reduction. The rats in the SA-I group also showed several functional (testosterone level and sperm parameter) and morphological (testicular weight and seminiferous tubule diameter) reductions. The rats in the SP-L group showed increased body weight and intertubular compartment volumetric and absolute densities and reduced tubular compartment volumetric density. The rats in the SA-L group presented only reduced sperm viability. Stress stimuli promoted changes in the rats in all the study groups. The testes of the adult rats were the most affected by chronic stress. However, the stressed adult rats recovered well from the testicular alterations.

Immunohistochemical and Cytoarchitectonic Studies of Testes in Triticum aestivum Linn (Poaceae) Lectin‐Administered Sprague‐Dawley Rats

This study investigated the reproductive sequelae of administration of common wheat lectin on the cytoarchitecture and immunohistochemically localized androgen receptors (AR) sites in prepubertal rat testes. Forty‐five male prepubertal rats weighing 50–60 g and approximately 30 days old were randomized into three groups represented by different time intervals of two, five and ten weeks, respectively. Each study time frame consisted of three sub‐groups of five rats each. Each sub‐group consisted of control which received 10 mL/kg BW/day of distilled water, reference standard group which received doses of 250 mL/kg BW of pure common wheat lectin while the third group received equal dose of common wheat lectin fraction. All administrations were given daily by gavage via oral cannula. At the end of each study time frame, the rats were sacrificed. The testes were excised and processed for immunohistochemical and cytoarchitectonic studies. Cytoarchitectural assessment of the testes of common wheat lectin exposed rats was characterized by varying degrees of tubular degenerations including germ cells depletion, segmental partial germ cell loss, exfoliation of germ cells into the lumen, appearance of residual bodies in the lumen as well as degeneration and distortion of interstitial (Leydig) cells. These changes were more significant in prepubertal rats administered with a dose of 250 mg/kg BW/day of pure common wheat lectin for 10 weeks. Common Wheat Lectin gradually decreased the androgen receptor expression in the Leydig, Sertoli and peritubular myoid cells, which could induce male factor infertility in rats. The results of this study indicated that lectins from Triticum aestivum Linn. disrupted reproductive parameters in prepubertal male rats time dependently through alteration of the testicular cytoarchitecture, markedly and gradually decreased the androgen receptor expression in the Leydig, Sertoli and peritubular myoid cells, which could induce male factor infertility in rats.

P-01-024 Evaluation of immediate and late effects of chronic stress on testes of prepubertal and adult rats

P-01-024 Evaluation of immediate and late effects of chronic stress on testes of prepubertal and adult rats

815 Evaluation of immediate and late effects of chronic stress on testes of prepubertal and adult rats

815 Evaluation of immediate and late effects of chronic stress on testes of prepubertal and adult rats

DBP-induced endoplasmic reticulum stress in male germ cells causes autophagy, which has a cytoprotective role against apoptosis in vitro and in vivo.

Recently, spermatogenic cell apoptosis was shown to play a key role in the induction of testicular atrophy by dibutyl phthalate (DBP), thus causing reproductive toxicology. However, the molecular events induced by DBP in apoptotic germ cells remain unclear. In the present study, the mouse spermatocyte-derived GC-2 cell line was exposed to different doses of DBP. We found that DBP induced marked apoptosis in GC-2 cells. The levels of the major endoplasmic reticulum (ER) stress markers GRP-78, ATF-6, and p-EIF2α were elevated when GC-2 cells were exposed to 25 μM DBP and increased in a dose-dependent manner at higher concentrations. Furthermore, at a concentration that resulted in significant apoptosis (100 μM), CHOP, which plays a convergent role in ER stress-mediated apoptosis and is regulated by various upstream ER stress signals, was activated and partially contributed to the DBP-induced apoptosis. However, inhibition of ER stress by 4-PBA, a chemical with chaperone-like activities, augmented the GC-2 cell apoptosis induced by DBP. Further experiments demonstrated that DBP-induced ER stress additionally had a protective role, mediated through the activation of autophagy. These results were confirmed in prepubertal rat testis germ cells; DBP treatment significantly induced testicular atrophy, accompanied by apoptosis, ER stress, and autophagy. Inhibition of ER stress and autophagy significantly aggravated the DBP-induced damage to the germ cells and testes. Taken together, our data suggest that DBP-induced ER stress in germ cells has a cytoprotective effect that is mediated through autophagy activation. These findings provide novel clues regarding the molecular events involved in DBP-induced germ cell apoptosis.

Butylbenzyl phthalate induces spermatogenic cell apoptosis in prepubertal rats.

Butylbenzyl phthalate (BBP), a suspected endocrine disruptor, adversely affects male reproductive function. In this study, morphological alterations of prepubertal rat testes caused by single administration of BBP, were examined by light microscopy. Three-week-old male rats were given a single dose of 500 mg/kg BBP by oral gavage and sacrificed at 3, 12, and 24 h after administration. Histopathological examination revealed progressive detachment and sloughing of spermatogenic cells into the lumen, and a significant increase in the number of TUNEL-positive (apoptotic) spermatogenic cells in the treated groups, compared to the control. Semithin sections confirmed the apoptotic cells by their prominent basophilia, condensed chromatin, and shrunken cytoplasm, hallmarks of apoptotic cell death. Immunohistochemistry identified disruption of Sertoli cell vimentin and actin filaments in the treated groups. To elucidate the recovery effects of BBP, rats were treated in the same way and were sacrificed at D1-12h after administration. The apoptotic index returned to normal at D9. While, the testes revealed lower weight gain until D12. These results show for the first time that BBP induces collapse of vimentin filaments in Sertoli cells which may lead to disruption of Sertoli-spermatogenic cell physical interaction and induces spermatogenic cell apoptosis.

Impact of 2.45 GHz microwave radiation on the testicular inflammatory pathway biomarkers in young rats: The role of gallic acid.

The aim of this study was to investigate electromagnetic radiation (EMR) transmitted by wireless devices (2.45 GHz), which may cause physiopathological or ultrastructural changes, in the testes of rats. We addressed if the supplemental gallic acid (GA) may reduce these adverse effects. Six-week-old male Sprague Dawley rats were used in this study. Forty eight rats were equally divided into four groups, which were named: Sham, EMR only (EMR, 3 hday-1 for 30 days), EMR + GA (30 mg/kg/daily), and GA (30 mg/kg/daily) groups. Malondialdehyde (MDA) and total oxidant status (TOS) levels increased (p = 0.001 for both) in EMR only group. TOS and oxidative stress index (OSI) levels decreased in GA treated group significantly (p = 0.001 and p = 0.045, respectively). Total antioxidant status (TAS) activities decreased in EMR only group and increased in GA treatment group (p = 0.001 and p = 0.029, respectively). Testosterone and vascular endothelial growth factor (VEGF) levels decreased in EMR only group, but this was not statistically significant. Testosterone and VEGF levels increased in EMR+GA group, compared with EMR only group (p = 0.002), and also increased in GA group compared with the control and EMR only group (p = 0.044 and p = 0.032, respectively). Prostaglandin E2 (PGE2 ) and calcitonin gene releated peptide (CGRP) staining increased in tubules of the testes in EMR only group (p < 0.001 for both) and decreased in tubules of the testes in EMR+GA group (p < 0.001 for all parameters). In EMR only group, most of the tubules contained less spermatozoa, and the spermatozoon counts decreased in tubules of the testes. All these findings and the regenerative reaction, characterized by mitotic activity, increased in seminiferous tubules cells of the testes in EMR+GA group (p < 0.001). Long term EMR exposure resulted in testicular physiopathology via oxidative damage and inflammation. GA may have ameliorative effects on the prepubertal rat testes physiopathology. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1771-1784, 2016.

High Doses of Caffeine during the Peripubertal Period in the Rat Impair the Growth and Function of the Testis.

Prenatal caffeine exposure adversely affects the development of the reproductive organs of male rat offspring. Thus, it is conceivable that peripubertal caffeine exposure would also influence physiologic gonadal changes and function during this critical period for sexual maturation. This study investigated the impact of high doses of caffeine on the testes of prepubertal male rats. A total of 45 immature male rats were divided randomly into three groups: a control group and 2 groups fed 120 and 180 mg/kg/day of caffeine, respectively, via the stomach for 4 weeks. Caffeine caused a significant decrease in body weight gain, accompanied by proportional decreases in lean body mass and body fat. The caffeine-fed animals had smaller and lighter testes than those of the control that were accompanied by negative influences on the histologic parameters of the testes. In addition, stimulated-testosterone ex vivo production was reduced in Leydig cells retrieved from the caffeine-fed animals. Our results demonstrate that peripubertal caffeine consumption can interfere with the maturation and function of the testis, possibly by interrupting endogenous testosterone secretion and reducing the sensitivity of Leydig cells to gonadotrophic stimulation. In addition, we confirmed that pubertal administration of caffeine reduced testis growth and altered testis histomorphology.

The effect of IBMX and hormones on gene expression by rat Sertoli cells

BackgroundSertoli cells (Sc) regulate spermatogenesis under the control of FSH and testosterone (T). Functional maturation of Sc for supporting the spermatogenic onset during pubertal development is prerequisite for male fertility. However, the effect of hormone driven maturational changes in Sc is not well known. Objectives and experimental modelIn this present study we have compared hormone induced gene expression of immature and mature Sc isolated from neonatal (9-days old) and prepubertal (18-days-old) rat testes, respectively, to investigate the developmental difference of hormone responsiveness of Sc during postnatal maturation as well as influence of 3-isobutyl-1-methylxanthine (IBMX), a nonspecific inhibitor of phosphodiesterase in primary culture of Sc. Results and conclusionOur results suggested that FSH responsiveness of Sc obtained from 18-days-old rats were more prominent in terms of augmentation of lactate, cAMP and gene transcription as compared to Sc from 9-days of age. Our result also indicated that although the use of IBMX in primary culture of Sc generates a better readout in terms of FSH induced cAMP response, the presence of such pharmacological agent mellows down FSH stimulated gene expression profile. Our data indicated further that immature Sc are capable of differentiating in vitro if cultured with continuous supplementation of FSH and T (in combination). Taken together, we also concluded that for accurate evaluation of the modulation of gene expression by hormones, use of IBMX should be avoided in primary cultures of Sc.