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Spermatogenesis being a highly dynamic processes is highly vulnerable to various stresses including heat stress. Though, the relationship between physiological temperature and male germ cells is certainly immense, the magnitude of spermatozoal damage after exposure to heat is evidently degree and dose dependent. Further, there are contradictory reports related to germ cells apoptosis in relation to temperatures. Thus, currently the dynamics of temperature and time dependence on germ cell apoptosis were studied by modulating the heat treatment strategies. It was observed that the rate of apoptosis increased initially then decreased with time. The DNA fragmentation in the 10,000×g supernatant of testis homogenate of rats that received heat treatment for 15-min, 30-min as well as 45-min treatment with 15-min intermittent period was found to be almost equal. In various heat treated animals, the apoptosis was found to be maximum after day-1 of treatments, which then followed a decreased pattern. These results indicate that there may be an initial induction of apoptosis in the germ cells, which later primed or programmed the other germ cells to activate protective mechanisms against heat induced DNA damage and thus protecting germ cell population to undergo apoptosis at later durations., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Birds obtained by embryo engineering are used to study embryo development and to produce transgenic birds. As this method of producing birds still generate strong emotions of the public opinion head ornaments, testes and semen characteristics of sex chimera roosters were examined to check whether they differ from chickens obtained by non-manipulated methods. Measurements of head ornaments, testes and semen were correlated with each other. Semen quality factor (SQF) was calculated, as well as the level of fluctuating asymmetry (FA) of bilateral traits (wattles and testes). Positive correlation was found for comb width and wattle length and comb thickness and sperm concentration. Semen characteristics and FA did not exceed the level encounter in other chicken lines. Results obtained indicate that germline chimeras are similar in appearance of secondary sexual traits, and semen and testes characteristics to chickens produced in non-manipulated way., (© 2018 Blackwell Verlag GmbH.)

The present study showed that RNA helicase Belle (DDX3) was required intrinsically for mitotic progression and survival of germline stem cells (GSCs) and spermatogonial cells in the Drosophila melanogaster testes. We found that deficiency of Belle in the male germline resulted in a strong germ cell loss phenotype. Early germ cells are lost through cell death, whereas somatic hub and cyst cell populations are maintained. The observed phenotype is related to that of the human Sertoli Cell-Only Syndrome caused by the loss of DBY (DDX3) expression in the human testes and results in a complete lack of germ cells with preservation of somatic Sertoli cells. We found the hallmarks of mitotic G2 delay in early germ cells of the larval testes of bel mutants. Both mitotic cyclins, A and B, are markedly reduced in the gonads of bel mutants. Transcription levels of cycB and cycA decrease significantly in the testes of hypomorph bel mutants. Overexpression of Cyclin B in the germline partially rescues germ cell survival, mitotic progression and fertility in the bel-RNAi knockdown testes. Taken together, these results suggest that a role of Belle in GSC maintenance and regulation of early germ cell divisions is associated with the expression control of mitotic cyclins., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Aim: Fertility post-orchidopexy is dependent on transformation of neonatal gonocytes (G) into adult dark spermatogonia at about 3 months, the same time as gonadotrophins stimulate androgen secretion. We examined how androgen blockade affects transformation of gonocytes to spermatogonial stem cells (SSC) during this period in patients with undervirilisation syndromes., Methods: Patients with undervirilisation syndromes (n=30, 1.5 weeks-16 years) underwent review of medical records, pathology reports, and H&E slides of testes (ethics HREC32164). Fluorescent immunohistochemistry against anti-Mullerian hormone (AMH, Sertoli cells), mouse VASA homologue (MVH, germ cells) and DAPI (nuclei) allowed the number of MVH-positive gonocytes/spermatogonial stem cells per seminiferous tubular cross-section (G/T or SSC/T) to be counted., Results: Gonocytes (MVH-positive cells in the tubular lumen) were present in 15/16 patients under 2 years old. SSC (MVH-positive cells on the tubule basement membrane) were present in 25/30 patients. With increasing age, the mean number of SSC/T decreased from ~4 to 0, and G/T decreased from ~1.5 to 0. SSC were present in CAIS and PAIS patients at 1.5 and 3.5 weeks old, respectively., Conclusions: Gonocytes transform into SSC earlier than expected in patients with undervirilisation syndromes. Lack of androgens may stimulate non-androgenic regulators to trigger transformation. Understanding how gonocytes transform may enable optimization of spermatogonial development to preserve fertility post-orchidopexy., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Testicular damage is a common clinical problem in diabetic individuals that severely affects the quality of life. The present study investigates the possible protective mechanisms of telmisartan, an angiotensin II-receptor antagonist in the germ cell of diabetic rat. Male SD rats were used and randomized into six groups: control, telmisartan control, diabetic control and diabetic group treated with telmisartan at the doses of 3, 6 and 12mg/kg/day, per oral for 4 weeks. Diabetes was induced by injecting a single dose of streptozotocin (STZ), (55mg/kg) dissolved in ice-cold 10mM citrate buffer; pH 4.4 and administered i.p. immediately after preparation to the SD rats. At the end of the study, rats were sacrificed and the levels of nitrite, superoxide, malondialdehyde (MDA), glutathione (reduced and peroxidase) and superoxide dismutase (SOD) were measured. Germ cell toxicity was evaluated by using sperm count, sperm comet assay, histology of testes and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Further to confirm the oxidative and nitrosative damage, immunohistological quantification of 8-oxo-dG (8-oxo-7,8-dihydro-2'-deoxyguanosine) and 3-nitrotyrosine were evaluated respectively. Results showed that telmisartan significantly restored the levels of nitrite, superoxide, malondialdehyde, and glutathione and superoxide dismutase in diabetic testes. Further, telmisartan significantly increased the sperm counts, reduced apoptotic cell death, sperm DNA damage, oxidative and nitrosative damage in diabetic rat. Western blot analysis showed that telmisartan reduced the testicular inflammation and cell death by down-regulating the expression of NF-κB, IL-6, TNF-α, p-ERK1/2, iNOS, caspase-3 and increasing the PPAR-γ expression. Results clearly indicate that telmisartan significantly reduced the both oxidative and nitrosative stress, inflammation and cell death in diabetic testes. The present results confirmed that telmisartan exhibited beneficial role in the germ cell of diabetic rat., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Hydrogen sulfide (H2S) is an endogenously produced signaling molecule that belongs to the group of gasotransmitters along with nitric oxide (NO) and carbon monoxide (CO). H2S plays a pivotal role in male reproductive processes. It is produced in various tissues and cells of the male reproductive system, including testicular tissue, Leydig and Sertoli cells, epididymis, seminal plasma, prostate, penile tissues, and sperm cells. This review aims to summarize the knowledge about the presence and effects of H2S in male reproductive tissues and outline possible therapeutic strategies in pathological conditions related to male fertility, e. g. spermatogenetic disorders and erectile dysfunction (ED). For instance, H2S supports spermatogenesis by maintaining the integrity of the blood-testicular barrier (BTB), stimulating testosterone production, and providing cytoprotective effects. In spermatozoa, H2S modulates sperm motility, promotes sperm maturation, capacitation, and acrosome reaction, and has significant cytoprotective effects. Given its vasorelaxant effects, it supports the erection of penile tissue. These findings suggest the importance and therapeutic potential of H2S in male reproduction, paving the way for further research and potential clinical applications. [ABSTRACT FROM AUTHOR]

Background: Inflammation-induced testicular damage is a significant contributing factor to the increasing incidence of infertility. Traditional treatments during the inflammatory phase often fail to achieve the desired fertility outcomes, necessitating innovative interventions such as cell therapy. Methods: We explored the in vivo properties of intravenously administered Sertoli cells (SCs) in an acute lipopolysaccharide (LPS)-induced inflammatory mouse model. Infiltrating and resident myeloid cell phenotypes were assessed using flow cytometry. The impact of SC administration on testis morphology and germ cell quality was evaluated using computer-assisted sperm analysis (CASA) and immunohistochemistry. Results: SCs demonstrated a distinctive migration pattern, importantly they preferentially concentrated in the testes and liver. SC application significantly reduced neutrophil infiltration as well as preserved the resident macrophage subpopulations. SCs upregulated MerTK expression in both interstitial and peritubular macrophages. Applied SC treatment exhibited protective effects on sperm including their motility and kinematic parameters, and maintained the physiological testicular morphology. Conclusion: Our study provides compelling evidence of the therapeutic efficacy of SC transplantation in alleviating acute inflammation-induced testicular damage. These findings contribute to the expanding knowledge on the potential applications of cell-based therapies for addressing reproductive health challenges and offer a promising approach for targeted interventions in male infertility. [ABSTRACT FROM AUTHOR]

In vivo antigenotoxic activity of BP-C2 composition (at doses of 60, 80, and 120 mg/kg) based on polyphenolic compounds derived from hydrolyzed lignin was evaluated in mouse germ cells. The BP-C2 composition dose-dependently reduced the aneugenic activity of topoisomerase II inhibitor etoposide in mouse oocytes without affecting the clastogenic activity of the genotoxicant. In mouse testicular cells, the BP-C2 composition reduced the DNA-damaging activity of the pro-oxidant genotoxicant dioxidine, but not etoposide. The cytoprotective activity of BP-C2 composition was revealed in relation to etoposide-induced cytotoxicity. [ABSTRACT FROM AUTHOR]

The spatiotemporal expression patterns of genes are crucial for maintaining normal physiological functions in animals. Conditional gene knockout using the cyclization recombination enzyme (Cre)/locus of crossover of P1 (Cre/LoxP) strategy has been extensively employed for functional assays at specific tissue or developmental stages. This approach aids in uncovering the associations between phenotypes and gene regulation while minimizing interference among distinct tissues. Various Cre-engineered mouse models have been utilized in the male reproductive system, including Dppa3-MERCre for primordial germ cells, Ddx4-Cre and Stra8-Cre for spermatogonia, Prm1-Cre and Acrv1-iCre for haploid spermatids, Cyp17a1-iCre for the Leydig cell, Sox9-Cre for the Sertoli cell, and Lcn5/8/9-Cre for differentiated segments of the epididymis. Notably, the specificity and functioning stage of Cre recombinases vary, and the efficiency of recombination driven by Cre depends on endogenous promoters with different sequences as well as the constructed Cre vectors, even when controlled by an identical promoter. Cre mouse models generated via traditional recombination or CRISPR/Cas9 also exhibit distinct knockout properties. This review focuses on Cre-engineered mouse models applied to the male reproductive system, including Cre-targeting strategies, mouse model screening, and practical challenges encountered, particularly with novel mouse strains over the past decade. It aims to provide valuable references for studies conducted on the male reproductive system. [ABSTRACT FROM AUTHOR]

Infertility is a frequent long-term adverse effect of cancer therapy for children. Compromised testicular functions in adolescence are frequent observations after chemotherapy and there are currently no well-established alternatives to avoid this damage. Antimetabolites such as 6-mercaptopurine (6-MP) are used to treat a variety of cancer; however, its treatment-associated adverse effects on the male reproductive functions are overlooked. Here, the molecular processes underlying 6-MP-induced male germ cell damage in juvenile Sprague–Dawley (SD) rats (3 weeks) have been investigated. Rats were administered with low (5 mg/kg), medium (10 mg/kg), and high (20 mg/kg) doses of 6-MP per orally either singly (1 week × 1 cycle) or intermittently (1 week treatment followed by 1 week remission period × 3 cycles). The toxicity was evaluated in terms of genotoxicity and testes- and sperm-related cellular and molecular parameters. Single cycle of exposure either produced mild or no toxic manifestations at the end of the 6th week. Intermittent cycles of exposure, particularly at the 10 and 20 mg/kg, decreased body and organ weights, increased micronucleated cells in the peripheral blood, induced oxidative/nitrosative stress, altered sperm chromatin quality, reduced serum testosterone and follicle stimulating hormone (FSH) levels, increased testicular structural aberrations, DNA damage, and apoptosis, and upregulated TNF-α expression and downregulated p-AMPK and β-catenin expressions. Conclusively, intermittent cycles of exposure at 10 and 20 mg/kg doses to the juvenile rats significantly induced oxidative stress, genotoxicity, and cellular and molecular perturbations in the testes and sperm of adult rats. [ABSTRACT FROM AUTHOR]

Busulfan is an anticancer drug known to cause serious damage to seminiferous tubules in the testes and deplete germ cells in human and animal models. The testicular artery is anastomosed with deferential and cremasteric arteries and is divided into capsular arteries, which give rise to the centripetal arteries and then recurrent arteries. The arterial blood in the testicular tissue is supplied by such a consequent system of arterial vessels, in order from the peripheral to the central area. As anticancer drugs are generally distributed throughout the whole body via the bloodstream and the running and distribution of arteries differ among the testicular areas, we hypothesized that the efficacy of busulfan differs in different testicular areas, particularly between the central and peripheral areas. In this study, busulfan was intraperitoneally injected at 40 mg/kg body weight into C57BL/6J male mice. After 28 days, in busulfan- treated mice, the diameters of seminiferous tubules were significantly higher in the central than in the peripheral area of the testes. The seminiferous tubular areas also significantly decreased in the peripheral areas compared with the central areas. The number of germ cells per seminiferous tubule was significantly higher in the central than in the peripheral area. Sertoli cell nuclei were detached into the lumen in the peripheral area. The number of Leydig cells was significantly lower in the peripheral areas. These data suggest that the effects of busulfan differ between the central and peripheral areas of the testis at 4 weeks after busulfan administration. [ABSTRACT FROM AUTHOR]

STUDY QUESTION Do different boys with different types of cryptorchidism exhibit different anogenital distances (AGDs)? SUMMARY ANSWER Length of AGD seemed to differ in different groups of patients with cryptorchidism. WHAT IS KNOWN ALREADY AGD, which is used as an indicator of prenatal androgen action, tends to be shorter in boys with cryptorchidism compared to unaffected boys. Shorter AGDs have also been reported in boys with hypospadias, in men with poor semen quality, and in men with testicular cancer. STUDY DESIGN, SIZE, DURATION A prospective descriptive cohort study was performed using data from consecutively selected boys with cryptorchidism (n = 169) operated in a single center over a period of 3 years (September 2019 to October 2022). PARTICIPANTS/MATERIALS, SETTING, METHODS AGD was measured in 169 infant boys, at 3 to 26 months of age, during anesthesia with a vernier caliper measuring the distance from the anus to the base of the scrotum (AGDAS) and from the anus to the anterior base of the penis (AGDAP) in two body positions according to the methods by 'The Infant Development and the Environment Study' (TIDES) and 'Cambridge Baby Growth Study', resulting in four mean values per patient (TIDES AGDAS/AP and Cambridge AGDAS/AP). Normal values for AGD by age were set by our hospital Department of Growth and Reproduction based on a large cohort of healthy infant boys (n = 1940). Testicular biopsies were performed at orchidopexy as a clinical routine. The germ cell number (G/T) and type Ad spermatogonia number (AdS/T) per cross-sectional tubule of at least 100 and 250 tubules, respectively were measured and related to normal samples. Blood samples were obtained by venipuncture for measuring serum LH, FSH, and inhibin B. They were analyzed in our hospital Department of Growth and Reproduction where the normal reference was also established. Correlations between the four mean AGD measurements for each boy were evaluated by Spearman rank correlation analyses. The AGD measurement of every boy was transferred to the multiple of the median (MoM) of the normal AGD for age and named MoM AGD. MAIN RESULTS AND THE ROLE OF CHANCE There were 104 boysoperated for unilateral, and 47 boys operated for bilateral, undescended testes, whereas 18 boys had vanished testis including one boy with bilateral vanished testes. Only 6% of cases with vanished testes had a MoM AGD higher than the normal median compared to 32% with undescended testes (P  < 0.05). MoM AGD increased with the age at surgery for boys with vanished testis (Spearman r  = 0.44), but not for boys with undescended testes (Spearman r  = 0.14). Boys with bilateral cryptorchidism had longer AGDs and more often had hypogonadotropic hypogonadism than boys with unilateral cryptorchidism (P  < 0.005) and (P  < 0.000001). LIMITATIONS, REASONS FOR CAUTION Although being the largest published material of AGD measurements of infant boys with cryptorchidism, one limitation of this study covers the quite small number of patients in the different groups, which may decrease the statistical power. Another limitation involves the sparse normal reference material on G/T and AdS/T. Finally, there are currently no longitudinal studies evaluating AGD from birth to adulthood and evaluating childhood AGD in relation to fertility outcome. Our study is hypothesis generating and therefore the interpretation of the results should be regarded as exploratory rather than reaching definite conclusions. WIDER IMPLICATIONS OF THE FINDINGS The study findings are in agreement with literature as the total included group of boys with cryptorchidism exhibited shorter than normal AGDs. However, new insights were demonstrated. Boys with vanished testis had shorter AGDs compared to unaffected boys and to boys with undescended testes. This finding challenges the current concept of AGD being determined in 'the masculinization programming window' in Week 8 to 14 of gestation. Furthermore, boys with bilateral cryptorchidism had longer AGDs and more often had hypogonadotropic hypogonadism than boys with unilateral cryptorchidism, suggesting that the lack of fetal androgen in hypogonadotropic hypogonadism is not that significant. STUDY FUNDING/COMPETING INTEREST(S) No external funding was used and no competing interests are declared. TRIAL REGISTRATION NUMBER The trial was not registered in an ICMJE-recognized trial registry. [ABSTRACT FROM AUTHOR]

Adropin, a multifaceted peptide, was identified as a new metabolic hormone responsible for regulating gluco-lipid homeostasis. However, its role in the testicular function is not yet understood. We aimed to investigate the localization and expression of adropin and GPR19 during different phases of postnatal development. Immunohistochemical study revealed the intense reactivity of adropin in the Leydig cells during all phases of postnatal development, while GPR19 showed intense immunoreactivity in the pachytene spermatocytes and mild immunoreactivity in Leydig cells as well as primary and secondary spermatocytes. Western blot study revealed maximum expression of GPR19 in pre-pubertal mouse testis that clearly indicates maximum responsiveness of adropin during that period. So, we hypothesized that adropin may act as an autocrine/paracrine factor that regulates pubertal changes in mouse testis. To examine the effect of adropin on pubertal onset, we gave bilateral intra-testicular doses (0.5 and 1.5 µg/testis) to pre-pubertal mice. Adropin treatment promoted testicular testosterone synthesis by increasing the expression of StAR, 3β-HSD, and 17β-HSD. Adropin also promoted germ cell survival and proliferation by upregulating the expression of PCNA and downregulating the Bax/Bcl2 ratio and Caspase 3 expression resulting in fewer TUNEL-positive cells in adropin-treated groups. FACS analysis demonstrated that adropin treatment not only increases 1C to 4C ratio but also significantly increases the 1C (spermatid) and 1C to 2C ratio which demarcates accelerated germ cell differentiation and turnover of testicular cells. In conclusion, adropin promotes steroidogenesis, germ cell survival, as well as the proliferation in the pre-pubertal mouse testis that may hasten the pubertal transition in an autocrine/paracrine manner. [ABSTRACT FROM AUTHOR]

Cryptorchidism in males constitutes a notable risk factor for both infertility and testicular cancer. Infertility in adulthood is closely linked to the germ cell status in childhood. Furthermore, the significance of germ cell status is important as more than 95% of all reported testicular malignancies are germ cell tumors. The review aims to elucidate the pathogenesis of germ cells in cryptorchid testes concerning their association with infertility and testicular malignancies. Impaired germ cell numbers are evident in cryptorchid testes even during antenatal and neonatal stages. In cryptorchidism there is a rapid decline in germ cell number within the first year of life, partially attributed to physiologic gonocyte apoptosis. Additionally, germ cells fail to differentiate normally during mini-puberty leading to reduced germ cell proliferation and delayed clearance of gonocytes from the seminiferous epithelium. Absence of germ cells in testicular biopsies occurs already 10 months of age and germ cell deterioration progressively worsens with approximately 50% of persisting cryptorchid testes lacking germ cells during puberty. The deficient germ cell maturation and proliferation leads to later infertility. Elevated temperature in the cryptorchid testes and also hormonal deficiency contribute to this phenomenon. Germ cell neoplasia in situ (GCNIS) originating during fetal development may manifest in rare cases associated with disorders of sexual development, chromosomal abnormalities in boys, specific syndromes, and teratomas that include cryptorchidism. In adults, the presence of GCNIS predominantly represents a new histology pattern before invasive germ cell cancer is demonstrated and is neither congenital nor related to abnormal gonocyte transformation. [ABSTRACT FROM AUTHOR]

Oxidative stress is one of the main causes of testicular spermatogenesis disorders. Resveratrol (RES) can inhibit oxidative stress levels, reduce germ cell damage, and promote spermatogenesis. To investigate the molecular mechanism by which RES promotes spermatogenesis, the effects of hydrogen peroxide (H2O2) on the pathological damage of mouse testis tissue were assessed. The expression of key genes associated with pyroptosis, apoptosis, and autophagy, and the effects of RES on this process were determined. Experimental mice were divided into three groups with intraperitoneally injection of H2O2 (3 mmol/kg/d), RES (100 mg/kg/d) + H2O2 (3 mmol/kg/d), and normal saline, respectively. The testicular tissues were collected, and the pathological changes were observed by hematoxylin and eosin staining. The ultrastructure was observed using transmission electron microscopy (TEM). The transcriptional levels of pyroptosis, apoptosis, and autophagy-related genes were measured and analyzed by fluorescence quantitative polymerase chain reaction. The results showed that, in the H2O2 group, the basal membrane of spermatogenic tubules was damaged, atrophic, and thin. Interstitial cells were fragmented, and the gap of spermatogenic tubules increased. Compared with the H2O2 group, the pathological damage of testicular tissue structure in the H2O2 + RES group was significantly ameliorated. TEM results showed no evident damage to the cell structure in the control group, while apoptosis, autophagy, and pyroptosis were observed in the testicular tissue of H2O2 and H2O2 + RES groups, where cell damage was less, but apoptotic bodies, autophagosomes, and pyroptosomes were observed. The transcription levels of key pyroptosis genes increased in the H2O2 group but decreased significantly after RES treatment. The results indicated that H2O2 could induce pyroptosis, while RES inhibited pyroptosis and led to the release of inflammatory factors by regulating the expression of genes in signaling pathways. After H2O2 injection, the transcript levels of key apoptosis genes in testicular tissue were lower than those in the control group, while RES promoted the occurrence of H2O2-induced apoptosis, to achieve the purpose of protecting itself from oxidative damage. H2O2 could promote autophagy in mouse testicular tissue. RES enhanced the level of H2O2-induced autophagy, removed the damaged cells in the testicular tissue, and protected the development of the body. This study provided the experimental basis for the study of RES in oxidative damage, pyroptosis, apoptosis, and autophagy, and theoretical guidance and new ideas for drug development and treatment of male reproductive diseases in clinical practice. [ABSTRACT FROM AUTHOR]

Deleted in azoospermia-like (DAZL) is essential for mammalian testicular function and spermatogenesis. To explore the molecular characterization, expression patterns, and cellular localization of the DAZL in Hezuo pig testes, testicular tissue was isolated from Hezuo pig at five development stages including 30 days old (30 d), 90 days old (90 d), 120 days old (120 d), 180 days old (180 d), and 240 days old (240 d). DAZL cDNA was first cloned using the RT-PCR method, and its molecular characterization was analyzed using relevant bioinformatics software. Subsequently, the expression patterns and cellular localization of DAZL were evaluated using quantitative real-time PCR (qRT-PCR), Western blot, and immunohistochemistry. The cloning and sequence analysis showed that the Hezuo pig DAZL cDNA fragment contained 888 bp open reading frame (ORF) capable of encoding 295 amino acid residues and exhibited high identities with some other mammals. The qRT-PCR and Western blot results indicated that DAZL was specifically expressed in Hezuo pig testes, and DAZL levels of both mRNA and protein were expressed at all five reproductive stages of Hezuo pig testes, with extremely significant higher expression levels in 90 d, 120 d, 180 d, and 240 d than those in 30 d (p < 0.01). Additionally, immunohistochemistry results revealed that DAZL protein was mainly localized in gonocytes at 30 d testes, primary spermatocytes, and spermatozoon at other developmental stages, and Leydig cells throughout five development stages. Together, these results suggested that DAZL may play an important role by regulating the proliferation or differentiation of gonocytes, development of primary spermatocytes and spermatozoon, and functional maintenance of Leydig cells in testicular development and spermatogenesis of Hezuo pig. Nevertheless, the specific regulatory mechanisms underlying these phenomena still requires further investigated and verified. [ABSTRACT FROM AUTHOR]

Glucocorticoid excess adversely affects male reproduction. This study evaluates effects of pharmacological doses of niacin on testicular structure and function in dexamethasone‐treated rats. Adult rats (48) were randomly assigned to 6 equal groups: (1) Negative control (NC): normal rats; (2) Positive control (PC): dexamethasone at 7 mg/kg/day by intraperitoneal injections for 7 days; groups 3–6 (N50, N100, N200, and N400): dexamethasone and concomitant treatment with niacin at 50, 100, 200, and 400 mg/kg/day by oral gavages. Testicular weight and volume of PC rats were significantly lower than the NC group (p <.05). Testicular volume of rats in the N50 and N200 groups was statistically similar to the NC group. Significant decreases in serum testosterone with a slight LH increase were detected in the PC group. Nacin at 50 mg/kg reversed serum testosterone to NC levels and increased serum LH concentration. Niacin only slightly increased epididymal spermatozoa number while all groups of niacin‐treated rats had significantly higher percentages of motile spermatozoa compared with the PC group. Hypospermatogenesis, germ cell degeneration and depletion, epithelial vacuolization, and degenerated Leydig cells were observed in PC rats. Lesions were relatively milder in niacin‐treated rats. Johnsen scores were also significantly higher in niacin‐treated rats. Niacin reduced apoptosis as shown by TUNEL assay. In conclusion, niacin administration at pharmacological doses dose‐dependently ameliorates the destructive effects of dexamethasone on sperm motility, Johnsen score, and testicular cell apoptosis in rats with the latter can be considered a decisive mechanism for its positive effects on testis. [ABSTRACT FROM AUTHOR]

The success of chemotherapy regimens has led to an increase in cancer survival rate over the last decades. Melphalan has been widely used for the treatment of several types of cancers despite its gonadotoxic effects. Due to its ability to cause mutations in the spermatogonial stem cells and spermatids, melphalan can exert a negative impact on male reproductive health in young cancer survivors. β‐aminoisobutyric acid (BAIBA), a myokine released by skeletal muscles, has been reported to have beneficial effects in diabetic nephropathy, cardiomyopathy and hepatic toxicity. However, the exact role of BAIBA in chemotherapy‐induced germ cell toxicity is still unexplored. The present study aims to determine the dose‐dependent (25, 50, and 100 mg/kg) effects of BAIBA on melphalan‐induced (1.5 mg/kg) germ cell toxicity in sprague−dawley (SD) rats. The evaluation parameters included quantification of oxidative stress biomarkers, sperm count, sperm motility and head morphology, sperm and testicular DNA damage, sperm mitochondrial membrane potential, ultrastructural changes in sperms, histological and protein expression studies in testes. Melphalan treatment significantly altered all the above‐mentioned parameters and the high dose (100 mg/kg) of BAIBA restored melphalan‐induced toxicity in a significant manner by exerting antioxidant, anti‐inflammatory and antiapoptotic effects. However, the medium dose (50 mg/kg) of BAIBA decreased the toxicity of melphalan and the low dose (25 mg/kg) of BAIBA failed to counteract the melphalan‐induced male germ cell toxicity as well as the peripheral blood micronucleus induction. The antioxidant, anti‐inflammatory and antiapoptotic role of BAIBA in melphalan‐induced gonadal damage is a novel finding in an experimental rat model. [ABSTRACT FROM AUTHOR]

The spatiotemporal expression patterns of genes are crucial for maintaining normal physiological functions in animals. Conditional gene knockout using the cyclization recombination enzyme (Cre)/locus of crossover of P1 (Cre/LoxP) strategy has been extensively employed for functional assays at specific tissue or developmental stages. This approach aids in uncovering the associations between phenotypes and gene regulation while minimizing interference among distinct tissues. Various Cre-engineered mouse models have been utilized in the male reproductive system, including Dppa3-MERCre for primordial germ cells, Ddx4-Cre and Stra8-Cre for spermatogonia, Prm1-Cre and Acrv1-iCre for haploid spermatids, Cyp17a1-iCre for the Leydig cell, Sox9-Cre for the Sertoli cell, and Lcn5/8/9-Cre for differentiated segments of the epididymis. Notably, the specificity and functioning stage of Cre recombinases vary, and the efficiency of recombination driven by Cre depends on endogenous promoters with different sequences as well as the constructed Cre vectors, even when controlled by an identical promoter. Cre mouse models generated via traditional recombination or CRISPR/Cas9 also exhibit distinct knockout properties. This review focuses on Cre-engineered mouse models applied to the male reproductive system, including Cre-targeting strategies, mouse model screening, and practical challenges encountered, particularly with novel mouse strains over the past decade. It aims to provide valuable references for studies conducted on the male reproductive system.

Context: Testicular torsion–detorsion results in loss of germ cells and infertility. Pentoxifylline has been shown to prevent tissue damage. Aims: To determine the effect of pentoxifylline on germ cell survival in torsion–detorsion induced apoptosis Methods: Twenty male mice were divided into four groups of five animals each: Control, T1 (Torsion–detorsion + single dose 100 mg/kg Pentoxifylline/IP), T2 (Torsion–detorsion + daily 20 mg/kg pentoxifylline/IP for 2 weeks, and T/D (Torsion–detorsion only). 35th day after torsion–detorsion, the left testes of all the animals were harvested for histological and biochemical analysis. Key results: Histomorpholoical analysis showed significant increase (P < 0.05) in seminiferous tubule diameter, Johnsen's score and germ cells of Control and T1 compared to T2 and T/D, with no significant difference (P > 0.05) in testis weight, sertoli, leydig and myoid cells. Tunnel assay showed significant increase (P < 0.05) in apoptotic cells of T/D and T2 animals compared to Control and T1. RT-PCR analysis showed significant high (P < 0.01) mRNA expression of Bax gene in T/D compared to T1 and T2 and significant increase (P < 0.05) of Bcl2 in Control, T1, T2 compared to T/D. Nrf2-ARE transcripts revealed significant increase (P < 0.05) in Control and T1 compared to T2 and T/D. Western blot showed significantly increased (P < 0.05) caspase-3 in T/D compared to Control, T1 and T2. Conclusion: Pentoxifylline promotes spermatogenesis and suppressed apoptosis induced by testicular torsion–detorsion. Implication: Pentoxifylline could serve as adjunct therapy to surgery in the treatment of torsion–detorsion induced germ cell apoptosis. Testicular torsion is the condition whereby the testis is twisted about the spermatic cord, thus, cutting off blood supply to the testis. It is a painful condition that often leads to infertility. Therefore, in this study, we determined the protective effect of pentoxifylline against testicular damage in the condition of testicular torsion and subsequent detorsion in mouse model. The result of this study showed that pentoxifylline aids the survival of germ cells and prevent testicular apoptosis. [ABSTRACT FROM AUTHOR]

Spermatogenesis has been intensely studied in rodents but remains poorly understood in humans. Here, we used single-cell RNA sequencing to analyze human testes. Clustering analysis of neonatal testes reveals several cell subsets, including cell populations with characteristics of primordial germ cells (PGCs) and spermatogonial stem cells (SSCs). In adult testes, we identify four undifferentiated spermatogonia (SPG) clusters, each of which expresses specific marker genes. We identify protein markers for the most primitive SPG state, allowing us to purify this likely SSC-enriched cell subset. We map the timeline of male germ cell development from PGCs through fetal germ cells to differentiating adult SPG stages. We also define somatic cell subsets in both neonatal and adult testes and trace their developmental trajectories. Our data provide a blueprint of the developing human male germline and supporting somatic cells. The PGC-like and SSC markers are candidates to be used for SSC therapy to treat infertility.

Autophagy, an evolutionarily conserved cell reprogramming mechanism, exists in all eukaryotic organisms. It is a fundamental and vital degradation/recycling pathway that removes undesirable components, such as cytoplasmic organelles, misfolded proteins, viruses, and intracellular bacteria, to provide energy and essential materials for organisms. The success of male reproduction depends on healthy testes, which are mainly composed of seminiferous tubules and mesenchyme. Seminiferous tubules are composed of Sertoli cells (SCs) and various germ cells, and the main functional part of mesenchyme are Leydig cells (LCs). In recent years, a large amount of evidence has confirmed that autophagy is active in many cellular events associated with the testes. Autophagy is not only important for testicular spermatogenesis, but is also an essential regulatory mechanism for the ectoplasmic specialization (ES) integrity of SCs, as well as for the normal function of the blood–testes barrier (BTB). At the same time, it is active in LCs and is crucial for steroid production and for maintaining testosterone levels. In this review, we expanded upon the narration regarding the composition of the testes; summarized the regulation and molecular mechanism of autophagy in SCs, germ cells, and LCs; and concluded the roles of autophagy in the process of spermatogenesis and testicular endocrinology. Through integrating the latest summaries and advances, we discuss how the role of autophagy is a double-edged sword in the testes and may provide insight for future studies and explorations on autophagy in male reproduction. [ABSTRACT FROM AUTHOR]

Diabetes mellitus is known as the "epidemic of the century" due to its global prevalence. Several pre-clinical and clinical studies have shown that male germ cell toxicity is one of the major consequences of diabetes mellitus. Although β-aminoisobutyric acid (BAIBA) has been shown to be advantageous in the diabetic nephropathy and cardiomyopathy, its specific role in the diabetes-induced testicular toxicity remains unknown. In this study, an attempt was made to elucidate the molecular mechanisms of BAIBA-mediated germ cell protection in diabetic rats. Adult male Sprague-dawley rats were subjected to either no treatment (control) or BAIBA (100 mg/kg; BAIBA control) or Streptozotocin (50 mg/kg; diabetic control) or low (25 mg/kg), medium (50 mg/kg) and high (100 mg/kg) doses of BAIBA in diabetic conditions. Significant alterations in sperm related parameters, oxidative stress and apoptotic biomarkers, pancreatic and testicular histology, DNA damage and changes in expression of proteins in testes were found in the diabetic rats. 100 mg/kg of BAIBA significantly reduced the elevated blood glucose levels (P ≤ 0.05), increased body weight (P ≤ 0.01 in the 4th week), lowered malondialdehyde (P ≤ 0.05) and nitrite levels (P ≤ 0.01), elevated testosterone (P ≤ 0.05) and FSH levels (P ≤ 0.05), increased sperm count and motility (P ≤ 0.01), decreased testicular DNA damage (P ≤ 0.001), improved histological features of pancreas and testes, decreased TUNEL positive cells (P ≤ 0.01), decreased RAGE (P ≤ 0.01) and Bax (P ≤ 0.05) expressions and increased SIRT1 (P ≤ 0.05) and Atg 12 (P ≤ 0.05) expressions in the testes. 50 mg/kg of BAIBA partially restored the above-mentioned parameters whereas 25 mg/kg of BAIBA was found to be insignificant in counteracting the toxicity. It is interesting to note that BAIBA protects male germ cell damage in diabetic rats by regulating the IGF-1/AMPK/SIRT-1 signaling pathway. • RAGE-induced ROS and apoptosis caused diabetes-induced testicular toxicity. • 100 mg/kg of BAIBA significantly decreased diabetes-induced germ cell toxicity. • BAIBA upregulated IGF-1/AMPK/SIRT-1 signaling in testes. • Downregulation of apoptosis and upregulation of autophagy protected germ cells. • BAIBA is a novel protectant against diabetes-induced male germ cell toxicity. [ABSTRACT FROM AUTHOR]

Melphalan significantly contributes to the increase in childhood cancer survival rate. It acts as a gonadotoxic agent and leads to testes damage, dysbalance in gonadal hormones, and impairment in the germ cell proliferation. Therefore, it might be a potent threat to male fertility in individuals who have undergone melphalan treatment during childhood cancer. However, the molecular mechanisms of melphalan-induced gonadal damage are not yet fully explored and they need to be investigated to determine the benefit-risk profile. In the present study, juvenile male SD rats were subjected to single and intermittent cycles of melphalan exposure in a dose-dependent (0.375, 0.75 and 1.5 mg/kg) manner. Methods of end-points evaluations were quantification of micronuclei formation in peripheral blood, sperm count, sperm motility and head morphology, sperm and testicular DNA damage, histological studies in testes, oxidative/nitrosative stress parameters. A single cycle of exposure at high dose (1.5 mg/kg) produced significant effect on micronuclei formation only after the first week of exposure, whereas failed to produce significant effect at the end of the sixth week. Intermittent cycles of exposure at the dose of 1.5 mg/kg produced significant alterations in all the parameters (micronuclei in peripheral blood, testes and epididymides weight and length, MDA, GSH and nitrite levels, sperm count and motility, sperm head morphology, testicular and sperm DNA damage, protein expression in testes and histological parameters). So, time of exposure as well as the amount of exposure (total dosage administered) is critical in determining the magnitude of the damage in germ cell risk assessment. [ABSTRACT FROM AUTHOR]

In nonclinical toxicity studies, stage-aware evaluation is often expected to assess drug-induced testicular toxicity. Although stage-aware evaluation does not require identification of specific stages, it is important to understand microscopic features of spermatogenic staging. Staging of the spermatogenic cycle in dogs is a challenging and time-consuming process. In this study, we first defined morphologic features for the eight spermatogenic stages in standard histology sections (H&E slides) of dog testes. For image analysis, we defined the key morphologic features of five stages/pooled stage groups (I-II, III-IV, V, VI-VII, and VIII). These criteria were used to develop a deep learning (DL) algorithm for staging of the spermatogenic cycle of control dog testes using whole slide images. In addition, a DL-based nucleus segmentation model was trained to detect and quantify the number of different germ cells, including spermatogonia, spermatocytes, and spermatids. Identification of spermatogenic stages and quantification of germ cell populations were successfully automated by the DL models. Combining these two algorithms provided color-coding visual spermatogenic staging and quantitative information on germ cell populations at specific stages that would facilitate the stage-aware evaluation and detection of changes in germ cell populations in nonclinical toxicity studies. [ABSTRACT FROM AUTHOR]

The biodiversity of marine fishes is threatened globally by climate change and other anthropogenic activities, particularly in coral reef ecosystems. We present a simple, field-hardy method to cryopreserve marine fish gonads, targeting spermatogonial cells (undifferentiated diploid germ cells) with the ultimate goal of permitting recovery of threatened species and populations via gonadal diploid germ cell transplantation technologies. The use of a simplified cryopreservation extender based on L-15 medium resulted in minimal decline in spermatogonial cell viability post-thaw. Moreover, we compared post-cryopreservation viability of sperm and spermatogonial cells from gonads cryopreserved with freshly prepared cryoprotectant and with cryoprotectant prepared in advance and stored at −20 °C. We found no significant difference, suggesting that these solutions may be prepared in advance and frozen, ready for later use. We urge conservation, academic, and regulatory agencies to cryobank fish gonads as part of their sample collection processes to support the biodiversity and security of valuable marine fish resources, alongside other restoration efforts. [ABSTRACT FROM AUTHOR]

The Sertoli cells of the testes play an essential role during gonadal development, in addition to supporting subsequent germ cell survival and spermatogenesis. Anti-Müllerian hormone (AMH) is a member of the TGF-β superfamily, which is secreted by immature Sertoli cells from the 8th week of fetal gestation. lnhibin B is a glycoprotein, which is produced by the Sertoli cells from early in fetal development. In people with a Difference or Disorder of Sex Development (DSD), these hormones may be useful to determine the presence of testicular tissue and potential for spermatogenesis. However, fetal Sertoli cell development and function is often dysregulated in DSD conditions and altered production of Sertoli cell hormones may be detected throughout the life course in these individuals. As such this review will consider the role of AMH and inhibin B in individuals with DSD. [ABSTRACT FROM AUTHOR]

Molecular mechanisms of seasonal changes in testicular morphology, histoarchitecture, and functions in birds have not been fully elucidated. The aim of the study was to examine the alternations in cell proliferation and apoptosis, these processes-related gene expressions, and gap junction protein (GJA1 gene , connexin 43 protein; Cx43) expression and localization in gander testes during annual stages. Testes from domestic ganders (n = 28) in the first reproduction season were obtained at five stages, i.e., prebreeding (PrB), peak of reproduction (PR), postbreeding (PoB), nonbreeding (NB), and onset of reproduction (OR). Males were kept under controlled breeding conditions. Testicular weight, morphometry, and histology (H&E staining) were evaluated, and the following parameters were tested: (1) number of proliferating (PCNA-positive) and apoptotic (TUNEL-positive) cells; (2) mRNA and protein abundances of PCNA, caspase-3, and Cx43 by qRT-PCR and Western blot, respectively; (3) activity of caspase-3 by fluorometric method; and (4) localization of Cx43 by immunofluoresence. Testicular weight was found to decrease by 4-fold at the NB stage with massive depletion of germ cells concomitantly with a reduction of seminiferous tubule (ST) diameter and ST lumen compared to the PR and OR stages. The number of proliferating germ cells was higher at PrB and PR than at the PoB stage, whereas the number of apoptotic cells was higher at PoB and NB compared to PrB and OR. Thus, proliferation-to-apoptosis ratios were lower in PoB and NB than other stages. Moreover, mRNA expression of PCNA exhibited down regulation in these stages compared to PrB and PR. Stage-dependent changes in the Cx43 mRNA level and in the localization pattern in the ST germinal epithelium were observed. Lower abundances of GJA1 transcript during NB and OR than at PoB the stage and irregular distribution of Cx43 protein located near the lumen of ST primarily at PrB and NB compared to the remaining stages were noted. These results suggested that during gander testis development, function, regression, and recrudescence, the interaction between processes of cell proliferation, apoptosis, and changes in the localization of Cx43 protein in the germinal epithelium occurred. The balance between these processes may determine the final functional activity or inactive stage of the testes connected with weight and histoarchitecture changes. • Molecular mechanisms of annual testicular remodeling stages in ganders were studied. • 2. Testicular stages are maintained by balance in proliferation-to-apoptosis ratio. • The presence of Cx43 abundance in ganders' testes was shown. • Testicular stage-dependent localization of Cx43 protein was found. [ABSTRACT FROM AUTHOR]

Simple Summary: The most common form of reproduction in mammals is seasonal reproduction. This ensures that offspring are born at the most suitable time for survival, due to the abundance of food and the optimal temperatures for early postnatal development. In males, one way to achieve this is to decrease or lose fertility over a given period. This loss is associated with a greater or lesser degree of spermatogenesis modification that affects both germ and Sertoli cells. This paper reviews the different cellular mechanisms that have been postulated in recent years to explain how the activity of the seminiferous epithelium decreases during the non-reproductive period. Testicular regression occurs during the non-breeding season in many mammals. This affects spermatogenesis, resulting in decreased or arrested activity. Both lead to a decrease or cessation in sperm production. In recent years, the cellular mechanisms that lead to infertility in males in non-reproductive periods have been studied in very different species of mammals. At the start of the present century, the main mechanism involved was considered as an increase in the apoptotic activity of germ cells during the regression period. The loss of spermatogonia and spermatocytes causes not only a decrease in spermatogenesis, but an arrest of the seminiferous epithelium activity at the end of regression. Recently, in some mammal species, it was found that apoptosis is the usual mechanism involved in epithelium activity arrest, although it is firstly atrophied by massive desquamation of the germ cells that are released from their binding with the Sertoli cells, and which are shed into the lumen of the seminiferous tubule. In other species, it has been shown that not only germ cell apoptosis, but also Sertoli cell apoptosis, including decreased proliferative activity, spermatophagy or autophagy, are involved in testicular regression. Furthermore, the most recent studies indicate that there are multiple patterns of seminiferous epithelium regression in seasonally breeding animals, which may not only be used by different species, but also by the same ones to reproduce in the best conditions, ensuring their survival. In conclusion, at this time, it is not possible to consider the existence of a paradigmatic cellular mechanism in the involution of the seminiferous epithelium applicable to all male mammals with seasonal reproduction, rather the existence of several mechanisms which participate to a greater or lesser extent in each of the species that have been studied to date. [ABSTRACT FROM AUTHOR]

Various types of stem cells are being researched upon to exploit their potential for regenerative medicine including pluripotent human embryonic stem (hES) cells derived from spare human embryos, induced pluripotent stem (iPS) cells by reprogramming somatic cells to a pluripotent state and multipotent mesenchymal stem/stromal cells (MSCs) obtained in vitro from multiple tissues. More than 50 independent groups have reported another novel population of pluripotent stem cells in adult tissues termed very small embryonic-like stem cells (VSELs). VSELs are developmentally linked to primordial germ cells, which rather than giving rise to the germ cells and later ceasing to exist, survive throughout life in multiple organs along with tissue-specific adult stem cells better described as lineage-restricted, tissue-committed progenitors with limited plasticity. VSELs survive total body irradiation in bone marrow, oncotherapy in the gonads, bilateral ovariectomy in the uterus and partial pancreatectomy in the pancreas of mice and participate in the regeneration of multiple organs under normal physiological conditions. VSELs and tissue-specific progenitor cells work together in a subtle manner, maintain life-long tissue homeostasis and their dysfunction leads to various pathologies including cancer. However, due to their quiescent state, VSELs have invariably eluded lineage-tracing studies reported so far. Present article reviews novel insights into VSELs biology and how VSELs enriched from GFP (green fluorescent protein) mice have enabled to delineate their role in various biological processes in vivo. VSELs biology needs to be understood in-depth as this alone will help evolve the field of regenerative medicine and win the war against cancer. [ABSTRACT FROM AUTHOR]

Integrins are transmembrane receptors expressed in all nucleated mammalian cells, critically involved in cell-matrix adhesion and cell-cell interactions that modulate many signalling cascades. It is assumed that integrins also provide essential functions of the reproductive system. In this study, we describe the detailed localization and distribution of αV integrin in the plasma membrane of bull sperm head and tail. Integrin αV was observed in the area of forming acrosome in developing sperm since the stage of round spermatids and persists in the acrosome during epididymal maturation and ejaculation till the acrosomal exocytosis. We detected CD9 and CD81 tetraspanins as the potential partners of αV integrin. Their similar staining pattern in testicular tissue suggested the involvement of these molecules in the tetraspanin web of "testisomes". Moreover, the complex of αV with β1 and β3 integrin subunits cannot be excluded at least in sperm. The presented findings contribute to understanding the mutual action of integrins and tetraspanins during sperm development and maturation. [ABSTRACT FROM AUTHOR]

Panax ginseng is known to potentiate the immune system via its antioxidant properties. Cadmium as an environmental pollutant has destructive effects on body organs especially the reproductive system. This study investigated the effects of P. ginseng extract against cadmium-induced testicular toxicity in rats. In this experimental study, 50 male adult rats were randomly divided into 5 groups: A significant decrease was observed in sperm count and viability in the cadmium group versus the control group. Ginseng extract could repair these defects. Increases in apoptotic germ cells due to cadmium were significantly reduced by ginseng extract. Our results demonstrated that P. ginseng extract reduces cadmium-induced testicular toxicity, via anti-oxidative and anti-apoptotic processes. [ABSTRACT FROM AUTHOR]

Fluoride, a ubiquitous environmental compound, carries significant health risks at excessive levels. This study investigated the reproductive toxicity of fluoride exposure during puberty in mice, focusing on its impact on testicular development, spermatogenesis, and underlying mechanisms. The results showed that fluoride exposure during puberty impaired testicular structure, induced germ cell apoptosis, and reduced sperm counts in mice. Additionally, the SOD activity and GSH content were significantly decreased, while MDA content was significantly elevated in the NaF group. Immunohistochemistry showed an increase in the number of cells positive for GRP78, a key ER stress marker. Moreover, qRT-PCR and Western blot analyses confirmed the upregulation of both Grp7 8 mRNA and protein expression, as well as increased mRNA expression of other ER stress-associated genes (Grp94 , chop , Atf6 , Atf4, and Xbp1) and enhanced protein expression of phosphorylated PERK, IRE1α, eIF2α, JNK, XBP-1, ATF-6α, ATF-4, and CHOP. In conclusion, our findings demonstrate that fluoride exposure during puberty impairs testicular structure, induces germ cell apoptosis, and reduces sperm counts in mice. ER stress may participate in testicular cell apoptosis, and contribute to the testicular damage and decreased sperm counts induced by fluoride. Fluoride exposure during puberty induces testicular germ cell apoptosis, impairs testicular structure, and reduces sperm count in mice. ER stress may participate in testicular cell apoptosis via activating JNK and CHOP, ultimately contributing to testicular damage and decreased sperm counts in mice caused by fluoride exposure during puberty. [Display omitted] • Fluoride exposure during puberty impairs testis development in mice. • Fluoride exposure during puberty decreases sperm counts in mice. • Fluoride triggers testicular germ cells apoptosis in mice. • ER stress involves in fluoride-induced testicular germ cells apoptosis. [ABSTRACT FROM AUTHOR]

Using histological and morphometric research methods, the features of the spermatogenesis process in the convoluted seminal tubules of the testes of male white rats are shown. As a biological test object, white mongrel sexually mature male rats weighing 200-250 grams were used in the work. The material of the study is the testes of white male rats. Based on the morphometric measurements carried out - counting different types of spermatogenic cells in one convoluted seminal tubule, a table of their ratio was compiled. In the course of the conducted studies, it was found out that during the period of sexual maturity of male white rats, the largest percentage of the total number of spermatogenic cells in the spermatogenic epithelium are the earliest forming germ cells - spermatogonia, and when studying the quantitative change of various types of spermatogenic cells in the convoluted seminal tubule of the testes of male white rats, the largest number are mature male germ cells - spermatozoa. [ABSTRACT FROM AUTHOR]

The role of quercetin and vitamin E treatment against streptozotocin (STZ)‐induced testicular abnormalities in diabetic rats and the possible mechanism of action they use for protection were investigated. Diabetes was induced by STZ (45 mg/kg i.p. once) and blood glucose was determined. Plasmatic insulin, testosterone, luteinising hormone and follicle‐stimulating hormone (FSH) were determined by ELISA. Levels of cytochrome c, caspase 3 and caspase 9 were evaluated by immunohistochemistry, while lesions were viewed by histology. Insulin played a role in testicular protection against male infertility through modulation of luteinising hormone (LH). This consequently increased Leydig and Sertoli cells and maturation of germ cells with the attached epididymis having abundant spermatozoa. The study showed a positive correlation in the levels of LH, FSH and testosterone; it was further established that all treatments normalised diabetes‐induced alterations. Treatment with quercetin and vitamin E resulted in 34% decrease of apoptogenic cytochrome c release. This protected the testes against excessive apoptosis by decreasing caspase 3 and caspase 9 activation by up to 30 and 28% respectively (p <.05). Histology also showed that treatment prevented testicular cell death. The findings show that quercetin/vitamin E possess free radical scavenging properties that protected against testicular damage in diabetes. This suggests the possibility of pharmaco‐therapeutic intervention. [ABSTRACT FROM AUTHOR]

Di-(2-ethylhexyl) phthalate (DEHP) is a male reproductive toxicant. This research is aimed at investigating the effect of pubertal DEHP exposure on testicular endoplasmic reticulum (ER) stress and germ cell apoptosis. Five-week-old male mice were orally administered with DEHP (0, 0.5, 50, or 500 mg/kg/day) for 35 days. Testis weight and sperm count were reduced in mice exposed to 500 mg/kg/day DEHP. The number of seminiferous tubules in stages VII–VIII, mature seminiferous tubules, was reduced and the number of seminiferous tubules in stages IX–XII, immature seminiferous tubules, was elevated in mice treated with 500 mg/kg/day DEHP. Numerous apoptotic germ cells were observed in mouse seminiferous tubules exposed to 50 and 500 mg/kg/day DEHP. Moreover, cleaved caspase-3 was elevated in mouse testes exposed to 500 mg/kg/day DEHP. In addition, Bcl-2 was reduced and Bax/Bcl-2 was elevated in mouse testes exposed to 500 mg/kg/day DEHP. Additional experiment showed that GRP78, an ER molecular chaperone, was downregulated in mouse testes exposed to 500 mg/kg/day DEHP. Testicular p-IRE-1α, p-JNK, and CHOP, three markers of ER stress, were upregulated in mice exposed to 500 mg/kg/day DEHP. These results suggest that pubertal exposure to high doses of DEHP induces germ cell apoptosis partially through initiating ER stress in testes. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To observe morphological changes in testicular tissue of adult male BALB/c mice after administration of Cannabis sativa extract and to investigate the protective role of vitamin-C against cannabis-induced histopathological changes. METHODS: Eighteen BALB/c mice (31-40 g) were divided into 3 groups. Group- I was control group, Group-II received oral cannabis extract (14 mg/kg) and Group-III received cannabis extract (14 mg/kg) orally along with vitamin-C (100 mg/kg) for 45 days. At the end of experiment, variations in body weight, testicular weight and histological changes in testicular tissues of mice were observed. RESULTS: Average increase in weight was 7.000±0.77 gm, 4.833±0.87 gm & 5.833±0.79 gm in group-I, group-II and group-III respectively. Weight reduction in right and left testis was 42.30±0.465mg and 42.45±0.424mg in group-I, 38.85±0.523 mg and 39.35±0.520 mg in group-II, and 42.38±0.465 mg and 42.23±0.424 mg in group-III animals respectively (p<0.01). Mean seminiferours epithelium thickness was 4.367±0.302 µm in group-II, 5.683±0.104 µm and 6.417±0.116 µm in Group-I and group-III respectively (p<0.001). Average germ cells count (cells/tubule) in group-II was reduced (256.25±6.58) as compared to group-I (373.50±4.063) and group-III (366.25±3.442). Histological changes observed after the treatment were decreased height of germinal epithelium, basal lamina disruption, seminiferous tubules shrinkage, scanty cytoplasm with shrunken nuclei and germinal epithelium detachment from basal lamina in group-II animals as compared to group-I and group-III. CONCLUSION: The cannabis administration significantly declines the weight, size of testes, germ cells count and the height of seminiferours epithelium in animals. Vitamin-C had a protective role on cannabis induced toxic changes in the testes of the mice. [ABSTRACT FROM AUTHOR]

Diabetes increases the possibility of germ cell damage, hypogonadism, and male infertility. Diabetic condition negatively impacts zinc (Zn) and selenium (Se) levels in the body. Zn and Se are among the most important trace elements involved in the regulation of redox reaction, antioxidants enzymes activities, and DNA expression in a germ cell. The present study aimed to elucidate the combined effects of Zn and Se treatment on diabetes-induced germ cell damage in male Sprague Dawley rats. Type 1 diabetes was induced by the single intraperitoneal (i.p.) injection of streptozotocin (55 mg/kg). Zn (3 mg/kg, i.p.) and Se (0.5 mg/kg, i.p.) were administered daily for 8 consecutive weeks. All the animals were provided with normal feed and water throughout the study. The effects on germ cell damage were evaluated by body weight, feed-water intake, organ weight, sperm count, motility, sperm head morphology, biochemical analysis, histology, immunohistochemistry, halo assay, germ cell comet assay, testes terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end-labeling (TUNEL) assay, sperm TUNEL assay, serum protein pattern analysis, and subcellular analysis using transmission electron microscopy. Further, the expressions of nuclear erythroid-derived related factor 2, catalase, glutathione peroxidase 4, and glutathione peroxidase 5 were carried out to ascertain the mechanism of protection. The present results demonstrated that 8 weeks combined treatment of Zn (3 mg/kg, i.p.) and Se (0.5 mg/kg, i.p.) reduced diabetes-induced germ cell damage. This study further highlighted that Zn and Se combination treatment might be a better strategy for the germ cell protection in diabetes and deserve further investigation. [ABSTRACT FROM AUTHOR]

Intersexual individuals have both female and male genital tissues. We have described a case of an intersexual pig, which was identified during veterinary inspection of pigs at slaughterhouse. This organ represented ovotestis. Half of its tissues was represented by ovarian tissue resembling ovary by polycystic ovary syndrome. Another half was represented by testicular tissue with signs of germ cell aplasia. These observations can contribute to understanding the expression of intersexuality and intersexuality-reduced infertility in pigs, whose should be withdrawn from commercial and reproductive breeds. The careful analysis of reproductive organs of pigs with veterinarian diagnosis "intersexuality" by various methods is recommended. [ABSTRACT FROM AUTHOR]

The current study was designed to determine the effects of dietary quercetin on epididymal sperm and testicular changes in male rabbits during summer heat stress (HS). Twelve adult male New Zealand white rabbits were submitted to summer heat conditions (temperature-humidity index 29.9±1.2). One group was fed a basal ration (BR; n = 6), whereas the other group was fed the same basal diet supplemented with quercetin (30 mg/kg d; Que-BR; n = 6) for 60 d, and both groups were sacrificed at the end of the experiment. Epididymis and testicles isolation was done for sperm, histopathology and apoptosis assessments. The results showed that quercetin improved epididymis weight, but did not affect other testicular dimensions except testicular length. A significant improvement was observed in epididymal sperm motility, concentration, kinematic parameters, viability, mitochondrial potential and acrosome integrity in Que-BR compared to the BR group. Lowered serum malondialdehyde level was observed in quercetin supplemented rabbits. Moreover, the quercetin supplementation maintained the interstitial stroma, seminiferous tubules architecture, germinal and Sertoli cells under HS, decreasing the apoptotic germ cell rate in seminiferous tubules. In conclusion, HS condition affects the sperm and testes configurations in rabbits and dietary quercetin minimises oxidative stress, which in turn protects the testes and sperm against HS induced damage. [ABSTRACT FROM AUTHOR]

Spermatogenesis and spermatozoa ultrastructure of the amphibian leech Batracobdella algira Moquin-Tandon, 1846 (Hirudinida: Glossiphoniidae) have been investigated by means of electron and fluorescent microscopy. In B. algira, there are seven pairs of testisacs (testes) that are located latero-ventrally throughout the body. Each testis contains numerous cysts with developing germ cells. The germ cells in a given cyst are in the same developmental stage (i.e., there are spermatogonial, spermatocytic, and spermatid cysts); however, there is no developmental synchrony between the cysts, and therefore, all of the developmental stages occur simultaneously in the same testis. In the cysts, each germ cell is connected to acentral cytoplasmic mass, the cytophore, by one intercellular bridge. The spermatozoa of the studied species conform to the general organization plan that is known for Hirudinida: they are filiform cells that are formed in sequence by an elongated and twisted acrosome that consists of an anterior and posterior acrosome, a fully condensed and helicoid nucleus, a midpiece composed of a single and twisted mitochondrion that is characteristically surrounded by an electron-dense sheath, and a flagellum with the conventional 9 × 2 + 2 axonemal pattern. Using a comprehensive approach, we compared our findings with the ultrastructural data that had been obtained from the spermatozoa of the other hirudinids that have been studied to date. Only minor differences in the length and shape of the studied organelles were found which seems to be connected with the different ways of insemination, specific properties of female reproductive tracts, and physiology of fertilization. Additionally, we studied the organization of the microtubular cytoskeleton in male germline cysts at consecutive stages of spermatogenesis using fluorescent and electron microscopy. By comparing the present data with those from Oligochaeta, Branchiobdellida, and Acanthobdellida, we found that only the presence of an anterior acrosome characterizes the true leeches and that, at present, should be regarded as an autapomorphic character of Hirudinida. Our results showed that the arrangement of the microtubules changed dynamically during spermatogenesis. [ABSTRACT FROM AUTHOR]

In all metazoan species, sperm is produced from germline stem cells. These self-renew and produce daughter cells that amplify and differentiate dependent on interactions with somatic support cells. In the male gonad of Drosophila melanogaster, the germline and somatic cyst cells co-differentiate as cysts, an arrangement in which the germline is completely enclosed by cytoplasmic extensions from the cyst cells. Notch is a developmentally relevant receptor in a pathway requiring immediate proximity with the signal sending cell. Here, we show that Notch is expressed in the cyst cells of wild-type testes. Notch becomes activated in the transition zone, an apical area of the testes in which the cyst cells express stage-specific transcription factors and the enclosed germline finalizes transit-amplifying divisions. Reducing the ligand Delta from the germline cells via RNA-Interference or reducing the receptor Notch from the cyst cells via CRISPR resulted in cell death concomitant with loss of germline cells from the transition zone. This shows that Notch signaling is essential for the survival of the germline stem cell lineage. [ABSTRACT FROM AUTHOR]

Cilia are evolutionarily conserved hair-like structures with a wide spectrum of key biological roles, and their dysfunction has been linked to a growing class of genetic disorders, known collectively as ciliopathies. Many strides have been made towards deciphering the molecular causes for these diseases, which have in turn expanded the understanding of cilia and their functional roles. One recently-identified ciliary gene is ARL2BP, encoding the ADP-Ribosylation Factor Like 2 Binding Protein. In this study, we have identified multiple ciliopathy phenotypes associated with mutations in ARL2BP in human patients and in a mouse knockout model. Our research demonstrates that spermiogenesis is impaired, resulting in abnormally shaped heads, shortened and mis-assembled sperm tails, as well as in loss of axonemal doublets. Additional phenotypes in the mouse included enlarged ventricles of the brain and situs inversus. Mouse embryonic fibroblasts derived from knockout animals revealed delayed depolymerization of primary cilia. Our results suggest that ARL2BP is required for the structural maintenance of cilia as well as of the sperm flagellum, and that its deficiency leads to syndromic ciliopathy. [ABSTRACT FROM AUTHOR]

As a result of a high-throughput in situ hybridization screening for adult mouse testes, we found that the mRNA for Tmco5 is expressed in round and elongating spermatids. Tmco5 belongs to the Tmco (Transmembrane and coiled-coil domains) gene family and has a coiled-coil domain in the N-terminal and a transmembrane domain in the C-terminal region. A monoclonal antibody raised against recombinant TMCO5 revealed that the protein is expressed exclusively in the elongating spermatids of step 9 to 12 and is localized to the manchette, a transiently emerging construction, which predominantly consists of cytoskeleton microtubules and actin filaments. This structure serves in the transport of Golgi-derived non-acrosomal vesicles. Moreover, induced expression of TMCO5 in CHO cells resulted in the co-localization of TMCO5 with β-tubulin besides the reorganization of the Golgi apparatus. Judging from the results and considering the domain structure of TMCO5, we assume that Tmco5 may have a role in vesicle transport along the manchette. [ABSTRACT FROM AUTHOR]

For sexually reproducing organisms, production of male or female gametes depends on specifying the correct sexual identity in the germline. In D. melanogaster, Sex lethal (Sxl) is the key gene that controls sex determination in both the soma and the germline, but how it does so in the germline is unknown, other than that it is different than in the soma. We conducted an RNA expression profiling experiment to identify direct and indirect germline targets of Sxl specifically in the undifferentiated germline. We find that, in these cells, Sxl loss does not lead to a global masculinization observed at the whole-genome level. In contrast, Sxl appears to affect a discrete set of genes required in the male germline, such as Phf7. We also identify Tudor domain containing protein 5-like (Tdrd5l) as a target for Sxl regulation that is important for male germline identity. Tdrd5l is repressed by Sxl in female germ cells, but is highly expressed in male germ cells where it promotes proper male fertility and germline differentiation. Additionally, Tdrd5l localizes to cytoplasmic granules with some characteristics of RNA Processing (P-) Bodies, suggesting that it promotes male identity in the germline by regulating post-transcriptional gene expression. [ABSTRACT FROM AUTHOR]

Stem cells rely on instructive cues from their environment. Alterations in microenvironments might contribute to tissue dysfunction and disease pathogenesis. Germline stem cells (GSCs) and cyst stem cells (CySC) in Drosophila testes are normally maintained in the apical area by the testicular hub. In this study, we found that reproduction leads to accumulation of early differentiating daughters of CySCs and GSCs in the testes of aged male flies, due to hyperactivation of Jun-N-terminal kinase (JNK) signaling to maintain self-renewal gene expression in the differentiating cyst cells. JNK activity is normally required to maintain CySCs in the apical niche. A muscle sheath surrounds the Drosophila testis to maintain its long coiled structure. Importantly, reproduction triggers accumulation of the tumor necrosis factor (TNF) Eiger in the testis muscle to activate JNK signaling via the TNF receptor Grindelwald in the cyst cells. Reducing Eiger activity in the testis muscle sheath suppressed reproduction-induced differentiation defects, but had little effect on testis homeostasis of unmated males. Our results reveal that reproduction in males provokes a dramatic shift in the testicular microenvironment, which impairs tissue homeostasis and spermatogenesis in the testes. [ABSTRACT FROM AUTHOR]

Active adult stem cells maintain a bipotential state with progeny able to either self-renew or initiate differentiation depending on extrinsic signals from the surrounding microenvironment. However, the intrinsic gene regulatory networks and chromatin states that allow adult stem cells to make these cell fate choices are not entirely understood. Here we show that the transcription factor DNA Replication-related Element Factor (DREF) regulates adult stem cell maintenance in the Drosophila male germline. A temperature-sensitive allele of DREF described in this study genetically separated a role for DREF in germline stem cell self-renewal from the general roles of DREF in cell proliferation. The DREF temperature-sensitive allele caused defects in germline stem cell self-renewal but allowed viability and division of germline stem cells as well as cell viability, growth and division of somatic cyst stem cells in the testes and cells in the Drosophila eye. Germline stem cells mutant for the temperature sensitive DREF allele exhibited lower activation of a TGF-beta reporter, and their progeny turned on expression of the differentiation factor Bam prematurely. Results of genetic interaction analyses revealed that Mi-2 and Caf1/p55, components of the Nucleosome Remodeling and Deacetylase (NuRD) complex, genetically antagonize the role of DREF in germline stem cell maintenance. Taken together, these data suggest that DREF contributes to intrinsic components of the germline stem cell regulatory network that maintains competence to self-renew. [ABSTRACT FROM AUTHOR]