Structure Of Testis Research Articles

Oxidative Stress and Energy Metabolism in Male Reproductive Damage from Single and Combined High-Power Microwave Exposure at 1.5 and 4.3Ghz

The effect of multi-frequency electromagnetic environments on male reproduction has attracted the medical community’s interest. Studies have investigated the effects and mechanisms of single-frequency microwave exposure on male reproduction, but comparative research on high-power microwave (HPM) composite and single exposure remains scarce. This study aimed to examine the effects and mechanisms of combined 1.5GHz and 4.3GHz microwave exposure on male reproduction. Male Wistar rats were exposed to 1.5GHz (L-band) and 4.3GHz (C-band) electromagnetic radiation for 15minutes. The four groups were: sham, 10mW/cm² L-band, 10mW/cm² C-band, and 5mW/cm² L-band and 5mW/cm² C-band compound. Assessments were made on the pathological structures of testes, sperm viability, serum sex hormones, oxidative stress, and energy metabolism levels after radiation. Exposure to 1.5GHz and 4.3GHz microwaves individually resulted in testicular tissue damage and reduced sperm quality. There was little difference between the damage caused by HPM composite and single exposure. The exposed groups showed histological and ultrastructural changes, with reduced spermatozoa viability, motility parameters, and serum testosterone, luteinizing hormone, follicle-stimulating hormone, and serum inhibin-B on days 1 and 7 after exposure. These tended to recover partially by day 14. Adenosine triphosphate content and lactate dehydrogenase and succinate dehydrogenase activities in the exposed testicular tissue decreased, corresponding to decreased superoxide dismutase activity and increased malondialdehyde content. Both single and combined exposure to L- and C-band HPM affect the male reproductive system. Exposure to single and compound HPM shows no significant difference in risks, with oxidative stress and energy metabolism disturbances playing key roles.

Multi-omics revealed activation of TNF-α induced apoptosis signaling pathway in testis of DEHP treated prepubertal male rat

Di-(2-ethylhexyl) phthalate (DEHP) exposure has been associated with male reproductive damage, but the mechanisms involved remain incompletely defined. This study aims to investigate the effects of DEHP exposure on the testes of prepubertal rats through an integrative analysis of metabolomics and transcriptomics, combined with molecular experiments. DEHP exposure resulted in decreased testis weight and increased oxidative stress level in the testis tissues of prepubertal male rats. Moreover, our findings showed a disordered testis structure, reduced spermatogenic and Sertoli cells as well as destruction of mitochondria structure in the testis tissues of DEHP-treated prepubertal male rats. Transcriptome function analysis together with metabolome function analysis indicated that spermatogenesis, apoptosis, inflammatory, lipid metabolism as well as DNA repair signaling pathway were enriched in the testis of DEHP-treated prepubertal male rats. The integrative omics analysis further suggested that TNF-α induced apoptosis played a crucial role in mediating the detrimental effects of DEHP exposure on the testis of prepubertal rats, which was validated by ELISA, Western blotting and Tunel assays. Validation experiments conducted in vitro using GC-2 cells corroborated these findings, demonstrating that mono-(2-ethylhexyl) phthalate (MEHP), the main active metabolite of DEHP, significantly inhibits cell proliferation and increases apoptosis via activating the TNF-α apoptosis pathway. Overall, these findings provided a novel mechanism of dysregulated spermatogenesis of DEHP exposure on the testes of prepubertal rats.

Effect of Pregnyl (HCG) on Histological Structure of Testes in Albino Mice

Pregnyl is a placental hormone, produced mainly during the entire period of pregnancy, making it the vital sign for detecting pregnancy. The current study shed light on effect of Pregnyl on histological structure of testes in Albino mice. The study included 15 mice, and animals are divided into two groups. First group included five mice and second group included ten mice. They were placed in separate cages with continuous monitoring and cleaning throughout the study. First group was injected with Pregnyl at a concentration of 0.1 mg. /kg and the second group at a concentration of 0.2 mg/kg for a period of 30 days. After the end of the experiment, the animals were sacrificed, dissected, and their testicles were removed for the purpose of histological study. The outcome of this study shows that the animals in the two empirical groups which treated by Pregnyl showed changes in thickness of seminiferous tubules of testis wall and their shrinkage, as their manifestation becomes wavy and offbeat. furthermore, atrophy was noted in a few of seminiferous tubules, and the sloughing and exhaustion of some germ cells and their accumulating in lumen of seminiferous tubules. Additionally, degeneration was observed in some Sertolic cells.

Histological Study of Development Structure of Testis and Epididymis in Gazelle Gazelle Subgutturosa

Objective: The aim of this study for focused the light on the development structure of the testis of gazelle. Materials and Method: 10 healthy male gazelle were used to study the histology structure of testis (5 pre- Puberty and 5 post- Puberty). Results: The testis is surrounded by a thick, irregular connective tissue capsule. Each lobule of the testis is composed of seminiferous tubules and interstitial tissue, and trabecular and interlobular septa divide the lobules apart. Neighboring the seminiferous tubules are Sertoli cells and spermatogenic cells. From spermatogenic cells, spermatozoa are produced. The first spermatogonia are small, spherical cells with black, spherical nuclei that reside on the basement membrane. Spermatogonia were earliest stages of the spermatogenic cells. Larger cells with frequently distinct chromatin are called primary spermatocytes. Secondary spermatocytes are rare because of their quick second meiotic division and the haploid spermatids that follow. Spermatids are round, pale-nucleated cells that are grouped close to the lumen of the seminiferous tubule. Prostatogenic cells outnumber Sertoli cells in the body. Oval or triangular in shape, the nucleus has a light coloration. In the connective tissue separating adjacent tubules, Leydig cells are present. Lines the epididymis are pseudostratified epithelium. Extensions of the cytoplasm into the lumen are called stereocilia. In different proportions, there is circular smooth muscle, and a connective tissue lamina propria supports the epithelium. The rete testis is connected to the ductus epididymidis by the efferent ductules, which are convolutions of tubules. Numerous valved veins were visible in the tunica albuginea, which covered the initial ascending portion of the epididymal head. Cuboidal epithelium was lining extratesticular rete testis. Three different cell types; migrating, basal, and columnar had epithelial membranes lining the efferent ductules. Many columnar, non-ciliated cells with many cytoplasmic vacuoles and tiny granules lined the epithelium of the first segment of the efferent ductules. Conclusion: In post-puberty age, the structure of the gazelle's testis exhibited a pattern resembling that of other ruminants; the seminiferous tubules contain adult sperm and a broad lumen.

Persistent Mullerian Duct Syndrome in an Adult Infertile Male: A Case Report

Persistent Müllerian duct syndrome is a rare autosomal recessive disorder of sex development characterized by the presence of Müllerian duct derived structures in a normally virilized, genotypical (46, XY) and phenotypical male. Here we describe a case of male partner of the couple who presented with primary subfertility and was diagnosed with persistent müllerian duct syndrome during investigations. Persistent Müllerian duct syndrome is a rare disease and in developing countries like Nepal, because of unaware of the disease condition and lack of proper health care facilities, such cases are diagnosed later in adult males. Therefore, this case highlights the importance of awareness and knowledge for early detection and treatment of such conditions to conserve fertility and prevent malignancy of testis and other remnant mullerian structures.

Effects of female-specific selection for reproductive investment on male fertility traits.

Despite sharing an autosomal genome, the often divergent reproductive strategies of males and females cause the selection to act in a sex-specific manner. Selection acting on one sex can have negative, positive, or neutral fitness consequences on the opposite sex. Here, we test how female-limited selection on reproductive investment in Japanese quail (Coturnix japonica) affects male fertility-related traits. Despite there being no difference in the size of males' testes from lines selected for high female reproductive investment (H-line) or low female reproductive investment (L-line), in both lines, the left testis had a greater volume of sperm-producing tissue. Since H-line females have a larger left-side restricted oviduct, this suggests a positive genetic correlation between male and female gonad function and that internal testis structure is a target of sexual selection. However, despite H-line males having previously been found to have greater fertilization success in a competitive scenario, we found little evidence of a difference between the lines in sperm number, motility, velocity, length, or the number of sperm that reached the ova. Precopulatory cues and/or the role of seminal fluid in sperm motility may thus be more likely to contribute to the H-line male fertilization advantage in this species.

Ciprofloxacin Affect Sperm Quality and Induced Testicular and Hepatic Injury in Male Mice

Ciprofloxacin (CPFx) is one of a second-generation fluoroquinolones antibiotic and is prescribed a lot for curing bacterial infection. The aim of this study was to investigate of impact CPFx on sperm characteristics, liver DNA and histological structure of liver and testes. Twenty-four male mice aged 7-10 weeks were divided into three groups of eight. The first group (control) was given distilled water. The second and third groups received CPFx with doses 500 and 750 mg/kg respectively. All treatments were administrated orally for 14 days. Three untreated females were put with one treated male for mating. At the end of dosing period, mice were killed, testes and liver were removed and weighed. Sperm parameters, testosterone level and liver DNA were measured. Histopathology of liver and testes was carried out. The results showed that CPFx significantly affected liver DNA, sperm motility and morphology as well as it caused histopathological alterations of hepatic and testicular tissues. Moreover, it led to a significant elevation in the percentage of dead fetuses and also deformity fetuses. The results indicate that CPFX has detrimental effects on sperm quality, liver DNA and histological structure of testes and liver; therefore, it should be used with great caution

Effect of quercetin and mirtazapine on spermatogenesis and testis structure in phenylhydrazine-induced hemolytic anemia mice: An experimental study

Anemia poses a significant healthcare challenge across different socioeconomic groups and can result in reproductive system damage through the generation of free radicals and lipid peroxidation. This study examines the protective effects of quercetin (QUE) and mirtazapine (MIR) against the reproductive damage caused by phenylhydrazine (PHZ) in mice. Fifty NMRI mice, aged 8–10 weeks with an average weight of 27.0 ± 2.0 g, were randomly divided into five groups. The control group (Group 1) received oral administration of 10 mL/kg/day of normal saline. Group 2 (PHZ group) received an initial intraperitoneal dose of 8 mg/100 g body weight of PHZ, followed by subsequent doses of 6 mg/100 g every 48 h. Group 3 received PHZ along with oral QUE at a dosage of 50 mg/kg/day. Group 4 received PHZ along with oral MIR at a dosage of 30 mg/kg/day. Group 5 received PHZ along with oral QUE at a dosage of 50 mg/kg/day and MIR at a dosage of 30 mg/kg/day. The treatment duration was 35 days. Sperm samples were collected from the caudal region of the epididymis post-euthanasia to assess the total mean sperm count, sperm viability, motility, DNA damage, and morphology. Testicular tissue was employed to quantify total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) concentrations, while serum levels of testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were analyzed. Additionally, various aspects, including testicular histopathology, oxidative enzyme levels, gene expression related to apoptosis and antiapoptotic pathways, and in vivo fertility index, were evaluated after 35 days. The QUE, MIR, and QUE + MIR groups showed less abnormal morphology and DNA damage, as well as better total and progressive sperm motility, motility characteristics, viability, and plasma membrane function compared to the PHZ group. QUE, MIR, and QUE + MIR administration increased TAC, SOD, and GPx activities in testicular tissue, while reducing MDA levels compared to the PHZ group. Furthermore, QUE, MIR, and QUE + MIR significantly reduced Bax, and caspase-3 expression levels, and increased Bcl-2 expression levels, compared to the PHZ group. Mice treated with QUE, MIR, and QUE + MIR exhibited an increased in vivo fertility index and plasma sex hormone levels compared to the PHZ group. These results show that QUE, MIR, and QUE + MIR might be able to improve the fertility index, boost the testicular antioxidant defense system, and control the death of germ cells. This could mean that they could be used to treat mice with PHZ-induced testicular damage.

Hypogonadotropic hypogonadism in male tilapia lacking a functional rln3b gene

Relaxin 3 is a neuropeptide that plays a crucial role in reproductive functions of mammals. Previous studies have confirmed that rln3a plays an important role in the male reproduction of tilapia. To further understand the significance of its paralogous gene rln3b in male fertility, we generated a homozygous mutant line of rln3b in Nile tilapia. Our findings indicated that rln3b mutation delayed spermatogenesis and led to abnormal testes structure. Knocking out rln3b gene resulted in a decrease in sperm count, sperm motility and male fish fertility. TUNEL detection revealed a small amount of apoptosis in the testes of rln3b−/− male fish at 390 days after hatching (dah). RT-qPCR analysis demonstrated that mutation of rln3b gene caused a significant downregulation of steroid synthesis-related genes such as cyp17a1, cyp11b2, germ cell marker gene, Vasa, and gonadal somatic cell marker genes of amh and amhr2. Furthermore, we found a significant down-regulation of hypothalamic-pituitary-gonadal (HPG) axis-related genes, while a significantly up-regulation of the dopamine synthetase gene in the rln3b−/− male fish. Taken together, our data strongly suggested that Rln3b played a crucial role in the fertility of XY tilapia by regulating HPG axis genes.

The Effect of Dietary Organic Copper and Zinc Trace Minerals on some Yield and Mineral Levels and Histological Structure of Testes

This study aims to investigate the effects of providing Cu and Zn minerals with an organic structure reduced by 25% compared to the recommended (NRC) inorganic value on parameters such as the age at which lambs achieve 50% sperm motility yield, some developmental parameters, testis histology, as well as serum, wool, and fecal mineral levels in lambs. The study involved 12 male lambs in the T1 group (organic minerals) and 11 in the T2 group (inorganic minerals) of the Kıvırcık breed. Lambs received minerals from mothers during the last month of fetal period and suckling, continuing individual feeding post-weaning. During individual feeding, T1 received 5.25 mg/kg DM copper-chelate and 15.0 mg/kg DM zinc-chelate, while T2 received 7 mg/kg DM copper-sulfate and 20 mg/kg DM zinc-sulfate. The mothers received identical mineral amounts in the last month of pregnancy and lactation. The ewes birthed offspring solely for the supply of experimental groups. Lambs, averaging about 18.5 kg, underwent bi-weekly electroejaculation, concluding the trial for those with 50% semen motility. Statistical analyses were carried out using the GLM method. No differences were observed between groups in the average age at which lambs achieve 50% sperm motility, live weight, scrotum, and testicular measurements at this age (P > 0.05). Histological analyses revealed no difference in tubule area between T1 and T2 groups (P > 0.05), but tubular epithelium height was greater in T1 (P < 0.01). End-of-trial serum copper, as well as weaning day and end-of-trial serum and fleece zinc mean values, did not differ between groups (P > 0.05). However, T1 had higher mean values for serum Cu on the weaning day (P < 0.01), fleece copper on the weaning day (P < 0.05) and at the end of the trial (P < 0.001). Additionally, the T1 group exhibited lower mean levels of fecal copper (P < 0.05) and fecal zinc (P < 0.001). In conclusion, despite organic copper and zinc levels being 25% lower in the examined parameters, comparable or improved results were achieved with inorganic copper and zinc.

Protective Effect of Black Rice Cyanidin-3-Glucoside on Testicular Damage in STZ-Induced Type 1 Diabetic Rats.

Diabetic testicular damage is quite a common and significant complication in diabetic men, which could result in infertility. The natural fertility rate of type 1 diabetes men is only 50% because of testicular damage. This research first aimed to explore the intervention effect of C3G on testicular tissue damage induced by diabetes. Here, a streptozotocin-induced type 1 diabetic rat model was established, and then C3G was administered. After 8 weeks of C3G supplementation, the symptoms of diabetes (e.g., high blood glucose, lower body weight, polydipsia, polyphagia) were relieved, and at the same time that sperm motility and viability increased, sperm abnormality decreased in C3G-treated diabetic rats. Furthermore, the pathological structure of testis was restored; the fibrosis of the testicular interstitial tissue was inhibited; and the LH, FSH, and testosterone levels were all increased in the C3G-treated groups. Testicular oxidative stress was relieved; serum and testicular inflammatory cytokines levels were significantly decreased in C3G-treated groups; levels of Bax, Caspase-3, TGF-β1 and Smad2/3 protein in testis decreased; and the level of Bcl-2 was up-regulated in the C3G-treated groups. A possible mechanism might be that C3G improved antioxidant capacity, relieved oxidative stress, increased anti-inflammatory cytokine expression, and inhibited the apoptosis of spermatogenic cells and testicular fibrosis, thus promoting the production of testosterone and repair of testicular function. In conclusion, this study is the first to reveal that testicular damage could be mitigated by C3G in type 1 diabetic rats. Our results provide a theoretical basis for the application of C3G in male reproductive injury caused by diabetes.

Evaluation of the Potential Protective Role of Galangin Associated with Gold Nanoparticles in the Histological and Functional Structure of Testes of Adult Male Albino Mice Administrated with Carbon Tetrachloride

Galangin (Gal) is a natural flavonoid sourced from the roots of the plant Albina Kalanga. It possesses a variety of pharmacological activities, such as antioxidant, anticancer, and anti-inflammatory. The current study aims to enhance the efficacy of galangin through the use of gold nanoparticles as a drug delivery system against CCl4-induced toxicity in the testicular tissue of male albino mice. Forty-two albino mice were divided into seven groups (6 mice/group), treatment with carbon tetrachloride solution for two weeks by intraperitoneal injection, (1 ml/kg) once a week for all groups except the control group, after which a group was injected with gold nanoparticles and two groups were injected with galangin in two concentrations, and two groups were injected with AuNPs + Gal conjugation solution with two concentrations. The animals' bodies were weighed and blood samples were obtained for testosterone hormone analysis and testicles for the purpose of weighing and histological study at the end of the experiments. The study showed that the testicular tissue of mice treated with CCl4 had a different pathology compared to the control group, the testosterone hormone levels in the CCl4 group were significantly higher than those of the control group. Results from the AuNPs+Gal group showed a significant reduction in the effects of CCl4 toxicity on mice testicles, with testosterone levels are returning to normal and histological testicle structures improving, the ratio of testicle weights to animal body weight in the group injected with AuNPs was significantly higher than that of the control and CCl4 groups. In conclusion, the results revealed that galangin combined with gold nanoparticles could effectively reduced the histological and functional tissue damages caused by CCl4, presenting a promising natural solution that needs to be further developed

Special Staining and Protein Expression of VEGF/EGFR and P53/NF-κB in Cryptorchid Tissue of Erhualian Pigs.

Erhualian pigs exhibit one of the highest reproductive rates globally, and cryptorchidism is a crucial factor affecting reproductive abilities of boars. This investigation focused on cryptorchid tissues from Erhualian pigs, where the histological structure of cryptorchidism was observed using specialized staining. In addition, protein expression of P53/NF-κB in cryptorchid tissues was assessed using Western blot and immunohistochemistry. In comparison to normal Erhualian testes, Masson's trichrome staining indicated a reduction in collagen fibers in the connective tissue and around the basal membrane of the seminiferous tubules in cryptorchid testes. Moreover, collagen fiber distribution was observed to be disordered. Verhoeff Van Gieson (EVG) and argyrophilic staining demonstrated brownish-black granular nucleoli organized regions in mesenchymal cells and germ cells. When compared to normal testicles, the convoluted seminiferous tubules of cryptorchids exhibited a significantly reduced number and diameter (p < 0.01). Notably, VEGF/EGFR and P53/NF-κB expression in cryptorchidism significantly differed from that in normal testes. In particular, the expression of VEGF and P53 in cryptorchid tissues was significantly higher than that in normal testes tissues, whereas the expression of EGFR in cryptorchid tissues was significantly lower than that in normal testes tissues (all p < 0.01). NF-κB expressed no difference in both conditions. The expressions of VEGF and NF-κB were observed in the cytoplasm of testicular Leydig cells and spermatogenic cells, but they were weak in the nucleus. EGFR and P53 were more positively expressed in the cytoplasm of these cells, with no positive expression in the nucleus. Conclusion: There were changes in the tissue morphology and structure of the cryptorchid testis, coupled with abnormally high expression of VEGF and P53 proteins in Erhualian pigs. We speculate that this may be an important limiting factor to fecundity during cryptorchidism.

Toxicity evaluation of Dolichandrone serrulata flower extract on vital and reproductive organs in adult male rats.

Although Dolichandrone serrulata flower (DSF) aqueous extract has been shown to possess pharmacological properties, its systemic toxicity has still to be evaluated. The present study aimed to investigate the sub-chronic toxicity effect of DSF extract on biochemical parameters and histological structures of liver, kidney, testis, and epididymis plus vas deferens. Adult male rats were administered DSF at 100, 300, and 600 mg/kgBW via oral gavage for 48 consecutive days while control rats received distilled water. At the end of the experiment, blood, liver, kidney, testis, and epididymis plus vas deferens samples were collected to determine any changes to serum biochemical components including ALT, ALP, and creatinine levels and histological structures. The results revealed no significant difference in body weight and food or water consumption between control and the DSF-treated groups. It was found that DSF significantly increases the weight of epididymis plus vas deferens, while the kidney and liver showed a decrease in the high dose group (P value < 0.05). Histological changes in these vital and reproductive tissues including fibrosis were not observed after administration but ALT, ALP, and creatinine levels were significantly altered in the treated groups (P value < 0.05). These altered levels, however, were still within normal ranges. In conclusion, these findings demonstrated that D. serrulata flower extract had no sub-chronic toxicity on vital and reproductive structures but slightly altered some liver and kidney functions.

Investigation of the morphological and histological features of the testes of pigeon (Columba livia domestica) in pre-puberty and post-puberty.

Morphological and histological examination of the testes can provide a suitable insight into the health of the reproductive system. The objective of the current study was to investigate the morphological and histological features of the testes of local pigeons (Columba livia domestica) at mature and immature stages of age. Two groups of collected specimens underwent macroscopic and microscopic investigation to evaluate and compare the main general properties of their testes. The findings indicated that the testis has an oval shape in both pre-puberty and post-puberty stages, situated on the inner side of the kidney towards the caudal extreme of the lungs. However, the left testis was bigger than those on the right side. In the pre-puberty stage group, the testicular parenchyma was small, and almost collapsed seminiferous tubules containing a single layer of Spermatogonia and Sertoli cells. In contrast, in the post-puberty stage, the parenchyma space between seminiferous tubules was small, and tubules adhered closely to each other. Also, mature cells including sertoli, spermatogonia, and spermatocytes were noticed to spread within the tubules. The change in the histological structure of testes before and after maturity may help to evaluate the complexity of the male reproductive system of pigeons and draw attention to the organization of sex hormones and the function of several types of cells within the testes.

Anti-oxidant and anti-apoptotic effects of royal jelly against polystyrene microplastic-induced testicular injury in mice.

In recent years, microplastics (MPs), which are novel environmental contaminants measuring 5 mm in diameter, have garnered considerable attention. However, information regarding substances that can mitigate the dangers of MPs for animals remains extremely limited. Ninety days were devoted to the exposure of mature male mice to royal jelly (RJ) and 2 µm virgin polystyrene microplastics (PS-MPs) in this study. Pre-implantation embryo development; the structure of testis tissue; the gonadosomatic index; sperm parameters; RNA damage in germinal cells; the anti-oxidant capacity of the entire testis; and the activity of anti-oxidant enzymes in serum and testicular tissue, including TAC, SOD dismutase, CAT, GSH, and MDA, histomorphometric indices of the testis (tubular differentiation index, spermatogenesis index, and repopulation index), steroidogenic foci, and the quantity of apoptosis were assessed in the testis, respectively, through the measurement of pro-apoptosis (p53, Bax, and Caspase-3) and anti-apoptosis (Bcl-2) factors, as well as Hsp70 mediator. The results indicate that concurrent administration of RJ can confer a protective effect on mice exposed to microplastics by maintaining the structure of mitochondria and enhancement of the anti-oxidant defense system. Furthermore, RJ co-treatment decreased apoptosis and oxidant/anti-oxidant status, enhanced pre-implantation embryo development, and improved sperm characteristics and RNA damage in germ cells. The data confirm that royal jelly could protect the testis structure against polystyrene microplastic-induced testicular injury through anti-oxidant and anti-apoptotic properties.

Towards a "Testis in a Dish": Generation of Mouse Testicular Organoids that Recapitulate Testis Structure and Expression Profiles.

The testis is responsible for sperm production and androgen synthesis. Abnormalities in testis development and function lead to disorders of sex development and male infertility. Currently, no in vitro system exists for modelling the testis. Here, we generated testis organoids from neonatal mouse primary testicular cells using transwell inserts and show that these organoids generate tubule-like structures and cellular organization resembling that of the in vivo testis. Gene expression analysis of organoids demonstrates a profile that recapitulates that observed in in vivo testis. Embryonic testicular cells, but not adult testicular cells are also capable of forming organoids. These organoids can be maintained in culture for 8-9 weeks and shows signs of entry into meiosis. We further developed defined media compositions that promote the immature versus mature Sertoli cell and Leydig cell states, enabling organoid maturation in vitro. These testis organoids are a promising model system for basic research of testes development and function, with translational applications for elucidation and treatment of developmental sex disorders and infertility.

Gss deficiency causes age-related fertility impairment via ROS-triggered ferroptosis in the testes of mice

Glutathione synthetase (GSS) catalyzes the final step in the synthesis of glutathione (GSH), a well-established antioxidant. Research on the specific roles of the Gss gene during spermatogenesis remains limited due to the intricate structure of testis. In this study, we identified pachytene spermatocytes as the primary site of GSS expression and generated a mouse model with postnatal deletion of Gss using Stra8-Cre (S8) to investigate the role of GSS in germ cells. The impact of Gss knockout on reducing male fertility is age-dependent and caused by ferroptosis in the testis. The 2-month-old S8/Gss−/− male mice exhibited normal fertility, due to a compensatory increase in GPX4, which prevented the accumulation of ROS. With aging, there was a decline in GPX4 and an increase in ALOX15 levels observed in 8-month-old S8/Gss−/− mice, resulting in the accumulation of ROS, lipid peroxidation, and ultimately testicular ferroptosis. We found that testicular ferroptosis did not affect spermatogonia, but caused meiosis disruption and acrosome heterotopia. Then the resulting aberrant sperm showed lower concentration and abnormal morphology, leading to reduced fertility. Furthermore, these injuries could be functionally rescued by inhibiting ferroptosis through intraperitoneal injection of GSH or Fer-1. In summary, Gss in germ cells play a crucial role in the resistance to oxidative stress injury in aged mice. Our findings deepen the understanding of ferroptosis during spermatogenesis and suggest that inhibiting ferroptosis may be a potential strategy for the treatment of male infertility.

The Protective effects of Vitamin E against alterations of rat testis structure induced by deltamethrin; histological, ultrastructure, and biochemical study

ABSTRACT Deltamethrin is a widely used synthetic pyrethroid pesticide. It causes reproductive toxicity. Aim of the work: it evaluates the impact of vitamin E in restoration of the testicular integrity of albino rats after toxicity induced by Deltamethrin. Thirty-six adult male albino rats were included, and they were further sub-divided into four experimental groups; Group A: six rats served as controls. Group B (Model): 10 rats equally divided into two sub-groups (B1): the rats received deltamethrin dissolved in oil in a dose of 0.6 mg/kg/daily by nasogastric gavage for 2 weeks. (B2): the rats received Deltamethrin in the same dose of group B1 for 1 month. Group C (Protected): 10 rats equally divided into two sub-groups (C1): the rats received deltamethrin orally 0.6 mg/kg/day concomitant with Vitamin E dissolved in 1 ml of corn oil in a dose 200 mg/kg/day by nasogastric gavage for 2 weeks. (C2): the rats received deltamethrin concomitant with Vitamin E in the same dose of group C1 for 1 month. Group D (Treatment): 10 rats received deltamethrin for 1 month followed by Vitamin E for another month in the same previously prescribed doses. Significant decreases in serum testosterone level, GSH, catalase activity, and significant increase in MDA in the deltamethrin-treated group were detected. Moreover, histological and ultrastructural examinations of the testis seminiferous tubules showed detrimental alterations in the deltamethrin group which were duration dependent. Vitamin E administration reversed such alterations. Vitamin E ameliorates the testicular dysfunction caused by Deltamethrin.

L-valine supplementation disturbs vital molecular pathways and induces apoptosis in mouse testes

The branched-chain amino acids (BCAAs: leucine, isoleucine and valine) are essential for animal growth and metabolic health. However, the effect of valine on male reproduction and its underlying molecular mechanism remain largely unknown. Here, we showed that l-valine supplementation (0.30% or 0.45%, water drinking for 3 weeks) did not change body and testis weights, but significantly altered morphology of sertoli cells and germ cells within seminiferous tubule, and enlarged the space between seminiferous tubules within mouse testis. l-valine treatment (0.45%) increased significantly the Caspase3/9 mRNA levels and CASPASE9 protein levels, therefore induced apoptosis of mouse testis. Moreover, gene expression levels related to autophagy (Atg5 and Lamb3), DNA 5 mC methylation (Dnmt1, Dnmt3a, Tet2 and Tet3), RNA m6A methylation (Mettl14, Alkbh5 and Fto), and m6A methylation binding proteins (Ythdf1/2/3 and Igf2bp1/2) were significantly reduced. Protein abundances of ALKBH5, FTO and YTHDF3 were also significantly reduced, but not for ATG5 and TET2. Testis transcriptome sequencing detected 537 differentially expressed genes (DEGs, 26 up-regulated and 511 down-regulated), involved in multiple important signaling pathways. RT-qPCR validated 8 of 9 DEGs (Cd36, Scd1, Insl3, Anxa5, Lcn2, Hsd17b3, Cyp11a1, Cyp17a1 and Agt) to be decreased significantly, consistent with RNA-seq results. Taken together, l-valine treatment could disturb multiple signaling pathways (autophagy and RNA methylation etc.), and induce apoptosis to destroy the tissue structure of mouse testis.