Testes Of Male Offspring Research Articles

Prenatal BPA exposure disrupts male reproductive functions by interfering with DNA methylation and GDNF expression in the testes of male offspring rats.

BPA is a ubiquitous environmental endocrine-disrupting chemical, and maternal exposure to BPA is associated with impaired male reproductive functions; however, the mechanisms remain to be elucidated. Glial cell line-derived neurotrophic factor (GDNF) plays an important role in maintaining normal spermatogenesis and fertility. However, the effect of prenatal BPA exposure on GDNF expression and its mechanism in the testis has not been reported. In this study, pregnant Sprague-Dawley rats were respectively exposed to 0, 0.05, 0.5, 5, and 50 mg/kg/day BPA via oral gavage from gestational day (GD) 5 to GD 19, with 6 pregnant rats in each group. ELISA, histochemistry, real-time PCR, western blot, and methylation-specific PCR (MSP) were used to detect the sex hormone levels, testicular histopathology, mRNA and protein expression of DNA methyltransferases (DNMTs) and GDNF, and the promoter methylation of Gdnf in the testes of male offspring at postnatal day (PND) 21 and PND 56. Prenatal BPA exposure increased body weight; decreased sperm counts and serum levels of testosterone (T), follicle-stimulating hormone (FSH), and luteinizing hormone (LH); and induced testicular histological damage, which indicated the damage of male reproductive function. Prenatal BPA exposure also upregulated Dnmt1 in 5 mg/kg group and Dnmt3b in 0.5 mg/kg group, but down-regulated Dnmt1 in 50 mg/kg group at PND 21. At PND 56, Dnmt1 was significantly increased in 0.05 mg/kg group but decreased in 0.5, 5, and 50 mg/kg groups, Dnmt3a was decreased, and Dnmt3b was markedly increased in 0.05 and 0.5 mg/kg groups but decreased in 5 and 50 mg/kg groups. The mRNA and protein expression levels of Gdnf were decreased markedly in 0.5 and 50 mg/kg groups at PND 21. And the methylation level of Gdnf promoter was significantly increased in 0.5 mg/kg group, but decreased in 5 and 50 mg/kg groups at PND 21. In conclusion, our study indicates that prenatal BPA exposure disrupts male reproductive functions, interferes with the expression of DNMTs, and decreases Gdnf expression in the testes of male offspring. Gdnf expression may be regulated by DNA methylation; however, the detailed mechanism needs to be further investigated.

Tetramethyl bisphenol A stimulates proliferation but inhibits fetal Leydig cell function in male rats by targeting estrogen receptor α after in utero exposure.

Tetramethyl bisphenol A (TMBPA) is a widely used flame retardant. TMBPA has been a toxic to Leydig cells in puberty, but it remains unclear whether TMBPA has a similar inhibitor effect on fetal Leydig cells (FLCs). This study reported morphological and functional alterations of FLCs in the testes of male offspring at birth after in utero exposure to TMBPA. Pregnant Sprague Dawley rats were dosed via continuous gavage of TMBPA (0, 10, 50, and 200 mg/kg/day) from gestational day 14 to 21. TMBPA markedly raised serum total testosterone level, testicular volume, and FLC number of male offspring at 200 mg/kg dose. The up-regulation of Insl3, Star, and Cyp11a1 mRNAs was observed after 200 mg/kg TMBPA exposure. After normalization to the number of FLCs, TMBPA significantly reduced Lhcgr and Hsd3b1 expressions at 10mg/kg, and Cyp17a1 at 200 mg/kg paralleling with their protein levels. TMBPA compromised the expression of Esr1, while increased the expression of Cdk2 and Cdk4 as well as their protein levels. TMBPA particularly increased the phosphorylation of AKT1 and AKT2 at 200 mg/kg. In conclusion, the present study suggests that TMBPA may promote FLC proliferation via ESR1-CDK2/4-AKT pathway, while inhibits the function of FLCs by reducing steroidogenic enzyme activity.

CORO1A regulates lipoprotein uptake in Leydig cells exposed to cadmium

Cadmium (Cd) is one of the most common environmental pollutants, which has a long biological half-life. Maternal Cd-exposure in the natural environment causes steroidogenesis defects resulting in spermatogenesis disorder in male offspring. For better understanding its underlying mechanism, we have employed iTRAQ to screen the differentially expressed protein and found that the expression of CORO1A and Cofilin 1 was up-regulated approximately 2 fold in Leydig cells of maternal Cd-exposure offspring. As the major source of steroid hormone, cholesterol is transported to cells via receptor-mediated endocytosis which relies on the remodel of cytoskeleton, then stores in lipid droplets (LDs). However, few studies have focused on the role of cytoskeleton in abnormal steroidogenesis. This study was performed to explore the role of CORO1A in androgen deficiency caused by Cd exposure and its involvement of low-density lipoprotein (LDL) uptake and effects on LDs. We found that Cd resulted in the up-regulation of CORO1A and Cofilin 1, and down-regulation of Profilin 1 in the testis of male offspring with maternal exposure. The structure of filamentous actin was broken, disordered and even crumpled up in Cd-treated R2C cells. F-actin disassembly led to a low uptake of LDL with a reduced number of LDs, followed by decreased total cholesterol and low progesterone production. When CORO1A was silenced, the expression of Cofilin 1 was down-regulated and Profilin 1 was up-regulated in Cd-treated R2C cells. The filamentous actin was rescued and the integrated cytoskeleton prompted LDL uptake, which resulted in the increased total cholesterol and high progesterone production. These findings highlight the crucial role of CORO1A as a cytoskeleton regulatory protein in steroidogenesis, which may help to better understand Cd-induced steroid hormone deficiency in children.

The impact of silver nanoparticles phytosynthesized with Viburnum opulus L. extract on the ultrastrastructure and cell death in the testis of offspring rats

AimTo investigate the effects of prenatal exposure to AgNPs obtained by green synthesis with Viburnum opulus L. extract on the testis in male offspring rats. Material and methodsTwo different doses of AgNPs (0.8 and 1.5 mg/kg b.w.) and vehicle (PBS) were administered to Wistar female rats on days 3–14 of gestation. At 6 weeks after birth, the ultrastructural changes in correlation with the amount of silver as well as the parameters of oxidative stress, inflammation and cell death mechanisms in the testis of male offspring were evaluated. ResultsAgNPs administered during pregnancy crossed the placental and testicular barriers and induced oxidative stress, DNA damage and autophagy as mechanism of cell toxicity. The markers of inflammation and apoptosis decreased after AgNPs exposure while the NFkB activation increased. TEM examination revealed important ultrastructural changes of Sertoli cells, numerous vacuoles and cytoplasmic changes suggestive of the cellʹs evolution towards necrosis. ConclusionPhytoreduced silver nanoparticles with polyphenols from Viburnum opulus L. fruit extract, administered during the embryological development of the male gonad, have testicular toxic effects in offspring even at 6 weeks after birth.

Морфофункціональні зміни сім’яників щурів, народжених від матерів зі змодельованою фетоплацентарною недостатністю і за її корегування

Nowadays, it is almost unknown how fetoplacental insufficiency of mothers affects the formation of the reproductive system in male offspring during puberty. Therefore, the aim of this work was to study the histological structure of the testes of male offspring born to mothers of different ages with fetoplacental insufficiency and to evaluate the effectiveness of a new pharmaceutical composition for the correction of pathological conditions of pregnancy. The experiment involved healthy mature female Wistar rats in young (3–4 months) and mature (8–10 months) reproductive age. Eight groups with 7 pregnant females in each one were formed: groups 1 and 2 — intact animals of young and mature reproductive age, respectively; groups 3 and 4 — females with experimental fetoplacental insufficiency of young and mature reproductive age; groups 5 and 6 — young and mature animals with experimental fetoplacental insufficiency and addition to food depending on the weight of animals from 11 to 19 days of pregnancy pharmaceutical composition. Groups 7 and 8 — young and mature animals with experimental fetoplacental insufficiency with addition a comparison drug Dipyridamole to the food. Modeling of fetoplacental insufficiency was performed by daily subcutaneous injection to females from the 12th to the 18th day of pregnancy 50% oil solution of carbon tetrachloride at a dose of 2 ml/kg body weight. Offspring were decontaminated on the 50th day of life (puberty) by rapid decapitation. Samples of male testicles were fixed in 10% formalin solution, performed on alcohols of increasing strength, poured into paraffin. Sections were made from the blocks, which were stained with hematoxylin and eosin. Qualitative assessment of histostructure was performed on sections of the body in rats. Summing up the results of histological analysis and morphometric parameters characterizing the condition of the testicles of rats, we can draw the following conclusions. In the offspring born to reproductively young females with fetoplacental insufficiency, inhibition of the rate of germ cell differentiation was found, while in the offspring born to reproductively mature females with fetoplacental insufficiency, the negative effects of this effect were leveled and the rate of spermatogenesis increased. When the pharmaceutical composition was administered to both young and mature pregnant females on the background of fetoplacental insufficiency, it activated the processes of proliferation and differentiation of spermatogenic epithelium in their offspring, which was reflected in the appearance of mature sperm in the seminal tubules. The comparison drug Dipyridamole, administered according to the same regimen to both young and mature pregnant females with fetoplacental insufficiency, also helped the 50-day-old offspring of these females to restore the rate of spermatogenesis at the level of the pharmaceutical composition.

The effects of maternal exposure to BPA during pregnancy on the male reproductive system and the testicular microRNA expression profile.

The effect of prenatal bisphenol A (BPA) exposure is increasingly concerned. We investigated the effect of maternal BPA exposure during pregnancy on male offspring and its potential mechanism. Thirty pregnant Sprague Dawley (SD) rats were randomly divided into exposed and control groups. At PND56, the number of sperm, luteinizing hormone, and testosterone in the BPA-exposed group decreased, and testicular tissue structure was damaged in offsprings. At GD20, the miRNA profile in the testis of male offspring was examined and the expression levels of 28 deregulated miRNAs were validated by qRT-PCR. We found that miR-361-5p, miR-203a-3p, and miR-19b-2-5p had significantly different expression levels in the testis. These results suggest that maternal exposure to BPA can lead to differential changes in progeny miRNAs, which will provide direction for future in-depth mechanisms of reproductive injury.

Cuscuta chinensis flavonoids alleviate bisphenol A-induced apoptosis of testicular cells in male mice offspring.

Bisphenol A (BPA) is a widespread environmental endocrine disruptor that has multiple effects on reproductive organ development. To investigate the effect of Cuscuta chinensis flavonoids (CCFs) on testicular apoptosis induced by BPA in male mice offspring, pregnant mice were administered intragastrically with BPA and CCF at gestation day (GD) 0.5-17.5. The testes of male offspring (F1 males) were collected at post-natal day (PND) 21 and PND 56 for the detection of related indicators. The results showed that compared with the BPA group, the testicular index in CCF groups was significantly increased at PND 21 (p<.01). For the mice of different concentrations of CCF groups, the expression levels of bax, caspase-9 and caspase-7 proteins were significantly decreased at PND 21 and PND 56, while the expression level of bcl-2 protein was significantly increased, and testicular apoptotic cells were also decreased significantly (p<.01 or p<.05). Forty mg/kg CCF has no significant difference compared with the control group. The results indicated that CCF could protect the testis development of F1 male mice by alleviating the apoptosis of testicular cells induced by BPA.

Roles of piRNAs in microcystin-leucine-arginine (MC-LR) induced reproductive toxicity in testis on male offspring

In the present study, we evaluated the toxic effects on the testis of the male offspring of MC-LR exposure during fetal and lactational periods. Pregnant females were distributed into two experimental groups: control group and MC-LR group which were exposed to 0 and 10 μg/L of MC-LR, respectively, through drinking water separately during fetal and lactational periods. At the age of 30 days after birth, the male offspring were euthanized. The body weight, testis index, and histomorphology change were observed and the global changes of piwi-interacting RNA (piRNA) expression were evaluated. The results revealed that MC-LR was found in the testis of male offspring, body weight and testis index decreased significantly, and testicular tissue structure was damaged in the MC-LR group. In addition, the exposure to MC-LR resulted in an altered piRNA expression profile and an increase of the cell apoptosis and a decrease of the cell proliferation in the testis of the male offspring. It was reasonable to speculate that the toxic effects on reproductive system of the male offspring in MC-LR group might be mediated by piRNAs through the regulation of the target genes. As far as we are aware, this is the first report showing that MC-LR could play a role in disorder of proliferative and cell apoptosis in the testis of the male offspring by the maternal transmission effect of toxicity.

Effects of maternal and lactational exposure to 2-hydroxy-4-methoxybenzone on development and reproductive organs in male and female rat offspring.

2-Hydroxy-4-methoxybenzophenone (HMB) is an ultraviolet (UV) absorbing compound used in many cosmetic products as a UV-protecting agent and in plastics for preventing UV-induced photodecomposition. HMB has been detected in over 95% of randomly collected human urine samples from adults and from premature infants, and it may have estrogenic potential. To determine the effects of maternal and lactational exposure to HMB on development and reproductive organs of offspring, time-mated female Harlan Sprague-Dawley rats were dosed with 0, 1000, 3000, 10,000, 25,000, or 50,000 ppm HMB (seven to eight per group) added to chow from gestation day 6 until weaning on postnatal day (PND) 23. Exposure to HMB was associated with reduced body and organ weights in female and male offspring. No significant differences were observed in the number of implantation sites/litter, mean resorptions/litter, % litters with resorptions, number and weights of live fetuses, or sex ratios between the control and HMB dose groups. Normalized anogenital distance in male pups at PND 23 was decreased in the highest dose group. Spermatocyte development was impaired in testes of male offspring in the highest dose group. In females, follicular development was delayed in the highest dose group. However, by evaluating levels of the compound in rat serum, the doses at which adverse events occurred are much higher than usual human exposure levels. Thus, exposure to less than 10,000 ppm HMB does not appear to be associated with adverse effects on the reproductive system in rats.

Fluoride-elicited developmental testicular toxicity in rats: Roles of endoplasmic reticulum stress and inflammatory response

Long-term excessive fluoride intake is known to be toxic and can damage a variety of organs and tissues in the human body. However, the molecular mechanisms underlying fluoride-induced male reproductive toxicity are not well understood. In this study, we used a rat model to simulate the situations of human exposure and aimed to evaluate the roles of endoplasmic reticulum (ER) stress and inflammatory response in fluoride-induced testicular injury. Sprague–Dawley rats were administered with sodium fluoride (NaF) at 25, 50 and 100mg/L via drinking water from pre-pregnancy to gestation, birth and finally to post-puberty. And then the testes of male offspring were studied at 8weeks of age. Our results demonstrated that fluoride treatment increased MDA accumulation, decreased SOD activity, and enhanced germ cell apoptosis. In addition, fluoride elevated mRNA and protein levels of glucose-regulated protein 78 (GRP78), inositol requiring ER-to-nucleus signal kinase 1 (IRE1), and C/EBP homologous protein (CHOP), indicating activation of ER stress signaling. Furthermore, fluoride also induced testicular inflammation, as manifested by gene up-regulation of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in a nuclear factor-κB (NF-κB)-dependent manner. These were associated with marked histopathological lesions including injury of spermatogonia, decrease of spermatocytes and absence of elongated spermatids, as well as severe ultrastructural abnormalities in testes. Taken together, our results provide compelling evidence that ER stress and inflammation would be novel and significant mechanisms responsible for fluoride-induced disturbance of spermatogenesis and germ cell loss in addition to oxidative stress.

Testicular development and fertility of mice treated prenatally with N-nitroso-N-ethylurea at various gestational stages.

N-nitroso-N-ethylurea (ENU) was injected intraperitoneally (i.p.) into ICR female mice at 50 mg/kg on day 8, 10, 12, or 16 of gestation (plug day = day 0). Male newborns treated prenatally with ENU were obtained. Body and testes weights of males were measured on postnatal days 4, 12, and 21 and at 12 weeks of age, as well as histopathological observation of their testes. The treated males were mated at 10 weeks to untreated females of the same strain. Subsequently, the plasma testosterone concentration in each group was determined by enzyme immunoassay. Body weight on postnatal day 4 in the group treated on day 16 of gestation was significantly lower compared to that of the control group. On postnatal day 21 and at 12 weeks of age, body weight was significantly lower in all groups treated with ENU compared to that of the control group. Among the embryonic stages tested, embryonic day 10 is the most susceptible to ENU insult with respect to the postnatal development of testes and epididymides, when judged by the relative weight at 12 weeks of age. The fertility of the male offspring was drastically impaired by the prenatal ENU treatment on embryonic day 10, followed by day 12, while the fertility of male offspring treated on embryonic days 8 and 16 was not affected. Histopathological sections of testes of male offspring treated with ENU on embryonic day 10 resulted in the most severe changes in the seminiferous tubules. The plasma testosterone concentration was drastically lower in male offspring treated on embryonic day 10 compared to the control level. These results demonstrate that the impaired fertility of the ENU-affected mice was the result of paucity of germ cells and that the critical period in the male mouse fetus with respect to the disturbance of postnatal testicular development and fertility was around embryonic day 10, which is the period of primordial germ cell migration.

Gonadal effects of fetal exposure to the azo dye Congo red in mice: Infertility in female but not male offspring

The present study describes the relationship between gonadal agenesis and fertility in male and female mice exposed in utero to the diazo dye Congo red (CR). Maternal CR treatment inhibited testicular and ovarian function in the offspring after oral administration of 1 or 0.5 g/kg/day on Gestational Days 8–12. The testes of male offspring from CR-exposed dams were small in size and contained hypospermatogenic seminiferous tubules. However, despite the fact that testis weight was reduced by more than 70% in some males, they displayed normal levels of fertility when mated to untreated females for over 10 months. In contrast, female offspring from CR-exposed dams produced only about half as many litters and pups as the control pairs did under longterm mating conditions. Histological examination of the ovaries revealed that subfertility was correlated with ovarian atrophy. Females lacking maturing follicles were considerably less productive (1.3 litters and 11.5 pups) than treated females with histologically normal ovaries (7.1 litters and 78.1 pups). In summary, prenatal exposure to the dye CR affects the gonads of both male and female offspring, but only the female offspring display reduced fertility.

Gonadal Effects of Fetal Exposure to the Azo Dye Congo Red in Mice: Infertility in Female but Not Male Offspring

The present study describes the relationship between gonadal agenesis and fertility in male and female mice exposed in utero to the diazo dye Congo red (CR). Maternal CR treatment inhibited testicular and ovarian function in the offspring alter oral administration of I or 0.5 g/kg/day on Gestational Days 8–12. The testes of male offspring from CR-exposed dams were small in size and contained hypospermatogenic seminiferous tubules. However, despite the fact that testis weight was reduced by more than 70% in some males, they displayed normal levels of fertility when mated to untreated females for over 10 months. In contrast, female offspring from CR-exposed dams produced only about half as many litters and pups as the control pairs did under long- term mating conditions. Histological examination of the ovaries revealed that subfertility was correlated with ovarian atrophy. Females lacking maturing follicles were considerably less productive (1.3 litters and 11.5 pups) than treated females with histologically normal ovaries (7.1 litters and 78.1 pups). In sum mary, prenatal exposure to the dye CR affects the gonads of both male and female offspring, but only the female offspring display reduced fertility.

Experimental congenital adrenocortical hyperplasia: persistent postnatal deficiency in activity of 3-beta-hydroxysteroid dehydrogenase produced in utero.

ABSTRACT Activity of 3β-hydroxysteroid dehydrogenase is persistently deficient in maternal rat adrenal tissues at least as long as 10 months after a single injection of a steroidal substrate analogue (2α-cyano-4,4,17α-trimethylandrost-5-ene-17β-ol-3-one) on either the 16th or 19th day of gestation. The adrenal glands of offspring of mothers treated on either day are hyperplastic, and have deficient activity of this enzyme with reduced inhibitor titer throughout postnatal development. Testes of male offspring of mothers treated on the 16th day fail to develop maturational changes, whereas those of mature males whose mothers were treated on the 19th day have tubular atrophy and hyperplastic Leydig cells with reduced enzyme activity. Maternal and fetal pituitary glands are persistently enlarged after treatment. Similar results are obtained with the inhibitor with either propylene glycol or oil as vehicle, and with either intramuscular or amniotic routes of administration. Thus, a persistent enzymatic defect ...