Number Of Sertoli Cells Research Articles

Gene co-expression networks contributing to reproductive development in Holstein-Friesian bull calves

Enhanced early life nutrition stimulates the functionality of the hypothalamic-pituitary–testicular (HPT) biochemical signalling axis, resulting in precocious reproductive development in bull calves. Additionally, there is evidence that peptides and hormones produced within adipose tissue depots are also central in mediating the effect of metabolic status with reproductive development. The objective of this study was to undertake gene co-expression analyses on transcriptional data of the HPT and adipose tissues derived from bull calves fed contrasting planes of nutrition up to 18 weeks of life. The relationship between networks of co-expressed genes in each tissue dataset with calf phenotypic data was also assessed using a Pearson correlation analysis. Phenotypic data were related to metabolic status (systemic concentrations of insulin, leptin, adiponectin and IGF-1) reproductive development (systemic concentrations of testosterone, FSH and LH) and markers of testicular development (seminiferous tubule diameter, seminiferous tubule lumen score, spermatogenic cells and Sertoli cells). In the hypothalamus, gene co-expression networks involved in biochemical signalling processes related to gonadotropin-releasing hormone (GnRH) secretion were positively associated (P < 0.05) with systemic concentrations of IGF-1 and insulin. Similarly, a network of gene transcripts involved in GnRH signalling in the anterior pituitary was positively associated (P < 0.05) with systemic concentrations of LH. In the testes and adipose tissues, networks of co-expressed genes implicated in cholesterol and fatty acid biosynthesis were positively associated (P < 0.05) with lumen score, Sertoli cell number, and stage of spermatogenesis. Additionally, gene co-expression networks significantly associated (P < 0.05) with both metabolic and reproductive trait data were found to be enriched (P < 0.05) for biological pathways related to energy production, cellular growth and proliferation, GnRH signalling and cholesterol biosynthesis across multiple tissues examined. Results from this study highlight networks of co-expressed genes directly associated with markers of enhanced metabolic status and subsequent earlier reproductive development. Furthermore, genes involved in biological processes mentioned above may hold potential for informing genomic selection breeding programmes for the selection of calves capable of displaying earlier reproductive development as a consequence of enhanced dietary intake.

Correlation Study between Immunohistochemical Expression of Testicular Inhibin and Seminal Fluid Analysis

The inhibin family of dimeric peptide hormones are produced in the testis and are postulated to have paracrine and autocrine roles in the regulation of steroidogenesis, Testis is a reproductive organ composed of the seminiferous tubules which is the site of spermatogenesis, while the Sertoli and Leydig cells are responsible for secreting hormone; (inhibin), to maintain spermatogenesis. Dissimilarity of activin expression and plasma level it can cause variation to the parameters (sperm count, motility, germinal layer width). Aim of study was evaluation of the inhibin immunohistochemical activity in testicular tissue and the association between the inhibin expression and spermatogenesis. This study was performed in Al-Nahrain University / College of Medicine from November 2020 to June 2021. A total of 20 adult male rabbit. All animals were examined and monitored in order to exclude unhealthy animal. The animals were euthanized by inhalation of chloroform. The animals’ behavior observed during the experiment, in addition, we measured the weights of animals before and after the experiment. Dissection of the Rabbit was done to obtain the testis. We collected seminal fluid by milking the epididymis and then seminal fluid analysis was performed by professional lab technician. The volume of testis was measured. The specimens are kept in 10% formalin for (24-48) hours for histological preparation. In comparison between the two groups of the study (A, B) showed a statistically significant differences in means of sperm count, motility of sperm, germinal layer width, Leydig cell number, and Sertoli cell number between them. Regarding the means of inhibin expression and plasma level in the parameters (sperm count, motility of sperm, germinal layer width) were suggestively advanced in group B compared to that in group A. In conclusion, the expression and level of inhibin subunits, is correlated with parameters (sperm count, motility, germinal layer width), Inhibin peptide hormone acts as inhibitor for FSH-pituitary secretion that in turn promote spermatogenesis. The study shown a highly significant differences of inhibin manufacture and action through spermatogenesis is occupied in the fine regulator of the seminiferous epithelium of the cycle. Also, this study showed a negative correlation between the parameters with inhibin.

Methamphetamine can induce alteration of histopathology and sex determination gene expression through the oxidative stress pathway in the testes of human post-mortem.

Methamphetamine is a recreational drug that can be taken ingestion orally, injected, smoked or snorted. Methamphetamine abuse may lead to male infertility. The purpose of this study was to evaluate the long-term effects of methamphetamine abuse on the sex reprogramming of human post-mortem testis. Testes were collected from the autopsies of methamphetamine users (n= 10) and healthy males (reference group) (n= 10). They were then taken for stereological studies and RNA extraction to evaluate the expressions of PCNA, DMRT1, SOX8, c-Kit, TNF-α, IL6 and FOXL2 genes. In addition, Reactive Oxygen Species (ROS) level and Glutathione Disulfide (GSH) were assessed. Autopsied testicular samples of methamphetamine revealed a significant reduction in stereological parameters and histopathological findings, suggesting methamphetamine as a practical approach to prevention strategies in reproductive medicine that can disrupt spermatogenesis. Moreover, the results indicated the expressions of the genes involved in testis function and male-to-female genetic reprogramming (PCNA, DMRT1, SOX8, c-Kit, TNF-α, IL6 and FOXL2) (16) as well as in increasing inflammation (TNF-α and IL-6). The results also showed a high level of ROS and a decrease in GSH activity. The results of SOX9 immunohistochemistry indicated a significant decrease in the expression of SOX9 as well as in the number of Sertoli cells in the methamphetamine group. Overall, the results suggested that methamphetamine abuse caused spermatogenesis disruption and genetic reprogramming, probably through oxidative stress and changes in the expression of sex-determining genes.

Combination of Vitamin C and E Improves Spermatogenesis of White Male Rat Model of Paradoxical Sleep Deprivation Stress

Paradoxical Sleep Deprivation (PSD) elevates glucocorticoid and Reactive Oxygen Species (ROS) levels that cause oxidative stress, trigger spermatogenic cells damage, and reduce the number of sertoli cells. Vitamin C and E are antioxidants that could prevent spermatogenesis damage by preventing free radical formation. The study aimed to determine the effect of single and combined doses of vitamin C and E in improving spermatogenesis of white male rats (Rattus norvegicus) model of PSD. This is experimental research with post-test only and control group design on 28 white male rats distributed into four groups, i.e. group I (control, PSD), group II (PSD+vitamin C), group III (PSD+vitamin E), and group IV (PSD+a combination of vitamin C and E). Testicular preparations were stained using Hematoxylin-Eosin staining, quantitative scores of spermatogenic cells were measured using the Johnsen method, and the number of sertoli cells was counted in 10 seminiferous tubules in each of the three testicular sections. Group I has the lowest mean of Johnsen score (5.27±0.28), and group IV has the highest mean score (8.95±0.62), while the Mann-Whitney test showed a significant difference (p&lt;0.05) between group II, III and IV compared to group I (control group). Mean sertoli cells number of group I is the lowest (10.66±1.04), and group IV has the highest sertoli cells number. The post-hoc LSD test showed a significant difference (p&lt;0.05) between group IV and the other groups. Thus, the combination of vitamins C and E improves Johnsen score and sertoli cells number of male rats (Rattus norvegicus) experiencing PSD.

Maintenance of Sertoli Cell Number and Function in Immature Human Testicular Tissues Exposed to Platinum-Based Chemotherapy-Implications for Fertility Restoration.

Background: Retrospective studies in adult survivors of childhood cancer show long-term impacts of exposure to alkylating chemotherapy on future fertility. We recently demonstrated germ cell loss in immature human testicular tissues following exposure to platinum-based chemotherapeutic drugs. This study investigated the effects of platinum-based chemotherapy exposure on the somatic Sertoli cell population in human fetal and pre-pubertal testicular tissues. Methods: Human fetal (n = 23; 14–22 gestational weeks) testicular tissue pieces were exposed to cisplatin (0.5 or 1.0 μg/ml) or vehicle for 24 h in vitro and analysed 24–240 h post-exposure or 12 weeks after xenografting. Human pre-pubertal (n = 10; 1–12 years) testicular tissue pieces were exposed to cisplatin (0.5 μg/ml), carboplatin (5 μg/ml) or vehicle for 24 h in vitro and analysed 24–240 h post-exposure; exposure to carboplatin at 10-times the concentration of cisplatin reflects the relative clinical doses given to patients. Immunohistochemistry was performed for SOX9 and anti-Müllerian hormone (AMH) expression and quantification was carried out to assess effects on Sertoli cell number and function respectively. AMH and inhibin B was measured in culture medium collected post-exposure to assess effects on Sertoli cell function. Results: Sertoli cell (SOX9+ve) number was maintained in cisplatin-exposed human fetal testicular tissues (7,647 ± 459 vs. 7,767 ± 498 cells/mm2; p > 0.05) at 240 h post-exposure. No effect on inhibin B (indicator of Sertoli cell function) production was observed at 96 h after cisplatin (0.5 and 1.0 μg/ml) exposure compared to control (21 ± 5 (0.5 μg/ml cisplatin) vs. 23 ± 7 (1.0 μg/ml cisplatin) vs. 25 ± 7 (control) ng/ml, p > 0.05). Xenografting of cisplatin-exposed (0.5 μg/ml) human fetal testicular tissues had no long-term effect on Sertoli cell number or function (percentage seminiferous area stained for SOX9 and AMH, respectively), compared with non-exposed tissues. Sertoli cell number was maintained in human pre-pubertal testicular tissues following exposure to either 0.5 μg/ml cisplatin (6,723 ± 1,647 cells/mm2) or 5 μg/ml carboplatin (7,502 ± 627 cells/mm2) compared to control (6,592 ± 1,545 cells/mm2). Conclusions: This study demonstrates maintenance of Sertoli cell number and function in immature human testicular tissues exposed to platinum-based chemotherapeutic agents. The maintenance of a functional Sertoli cell environment following chemotherapy exposure suggests that fertility restoration by spermatogonial stem cell (SSC) transplant may be possible in boys facing platinum-based cancer treatment.

The role of ALOX15B in heat stress-induced apoptosis of porcine sertoli cells

Heat stress reduces the number of Sertoli cells and impairs spermatogenesis. Mounting evidence indicates that lipid homeostasis is fundamental to cell survival. However, little is known about the role of lipid peroxides in the heat stress-induced apoptosis of Sertoli cells. In the present study, we used metabolomics to explore the changes of lipid peroxides in porcine Sertoli cells under heat stress using liquid chromatograph-mass spectrometry. These results showed a notable increase in the content of 8-hydroxyeicosatetraenoic acid (8-HETE), and 15-hydroxyeicosatetraenoic acid (15-HETE). Furthermore, we found that among arachidonate lipoxygenases, heat stress significantly increased the expression of arachidonate 15-lipoxygenase type B (ALOX15B). Moreover, baicalein, a specific inhibitor of ALOX15B, reduced the content of 8-HETE and 15-HETE, and decreased the apoptosis rate in heat stress-treated porcine Sertoli cells. In addition, baicalein and small interfering RNAs targeting ALOX15B increased the content of 8-HETE and 15-HETE, and activated the p38-p53 pathway, causing apoptosis in heat stress treated porcine Sertoli cells. Interestingly, a p38 inhibitor decreased the expression of ALOX15B, reduced the content of 8-HETE and 15-HETE, and decreased the expression of p53 and the apoptosis rate in heat stress treated porcine Sertoli cells. A p53 inhibitor had similar effect on Sertoli cells. These results indicated that heat stress enhanced the expression of ALOX15B, increased the content of 8-HETE and 15-HETE, and activated the p38-p53 pathway to cause apoptosis. ALOX15B and lipid peroxides obtained feedback from the p38-p53 pathway. Our findings will help to reveal the mechanism of lipid metabolism in Sertoli cells, and could provide a new targeted substrate for anti-heat stress strategies.

An experimental approach to comprehend the influence of platelet rich growth factors on spermatogenesis

Leukocyte-poor platelet-rich plasma (LP-PRP) has a potential influence on tissue regeneration by stimulating the proliferation and differentiation of progenitor cells. The aim of this work is the morphological assessment of spermatogenesis under the influence of growth factors after electron irradiation. Materials and methods Experimental animals (n = 135) were divided into 5 groups: I – control (n = 10); II – 2IR (n = 35; 2 Gy); III – 2IR + LP-PRP + IGF-1 (n = 30); IV – 2IR + LP-PRP (n = 30); V – LP-PRP (n = 30). Results Electron irradiation reduces the number of germ cells in comparison with the control group. After injection of LP-PRP + rhIGF-1 significantly increased the number of germ cells, Sertoli and Leydig cells, the height of germinal epithelium, area and diameter of seminiferous tubules. Conclusion LP-PRP + rhIGF-1 has a normalizing effect on structural and functional disorders of the testis caused by electron irradiation.

Associations between testicular development and fetal size in the pig

BackgroundImpaired reproductive performance is the largest contributing factor for the removal of boars from commercial systems. Intrauterine growth restricted piglets represent 25% of the total number of piglets born and have impaired reproductive performance. This study aimed to improve the understanding of temporal changes in testicular gene expression during testes development in fetuses of different size. The lightest and closest to mean litter weight (CTMLW) male Large White × Landrace littermates were collected at gestational days (GD) 45, 60 and 90 (n = 5–6 litters/GD).ResultsTestes weight and testes weight as a percentage of fetal weight were not associated with fetal size at GD60 or 90. Fetal plasma testosterone was not associated with fetal size at GD90. There was no association between fetal size and seminiferous tubule area and number, number of germ or Sertoli cells per tubule. The lightest fetuses tended to have wider seminiferous tubules compared to the CTMLW fetuses at GD90 (P = 0.077). The testicular expression of KI67 (P ≤ 0.01) and BAX:BCL2 ratio (P = 0.058) mRNAs decreased as gestation progressed. Greater SPP1 mRNA expression was observed at GD60 when compared with GD45 and 90 (P ≤ 0.05). Lower expression of DMRT1 and SPP1 (P < 0.01) mRNAs was observed in testes associated with the lightest fetuses compared to the CTMLW fetuses at GD90.ConclusionsThese findings provide novel insights into the expression profiles of genes associated with testicular development and function. Further, these data suggest that programming of reproductive potential in IUGR boars occurs late in gestation, providing a platform for further mechanistic investigation.

Soy Formula Is Not Estrogenic and Does Not Result in Reproductive Toxicity in Male Piglets: Results from a Controlled Feeding Study.

Soy infant formula which is fed to over half a million infants per year contains isoflavones such as genistein, which have been shown to be estrogenic at high concentrations. The developing testis is sensitive to estrogens, raising concern that the use of soy formulas may result in male reproductive toxicity. In the current study, male White-Dutch Landrace piglets received either sow milk (Sow), or were provided milk formula (Milk), soy formula (Soy), milk formula supplemented with 17-beta-estradiol (2 mg/kg/d) (M + E2) or supplemented with genistein (84 mg/L of diet; (M + G) from postnatal day 2 until day 21. E2 treatment reduced testis weight (p < 0.05) as percentage of body weight, significantly suppressed serum androgen concentrations, increased tubule area, Germ cell and Sertoli cell numbers (p < 0.05) relative to those of Sow or Milk groups. Soy formula had no such effects relative to Sow or Milk groups. mRNAseq revealed 103 differentially expressed genes in the M + E2 group compared to the Milk group related to endocrine/metabolic disorders. However, little overlap was observed between the other treatment groups. These data suggest soy formula is not estrogenic in the male neonatal piglet and that soy formula does not significantly alter male reproductive development.

Effects of postnatal exposure to tetrabromobisphenol A on testis development in mice and early key events.

Whether or not tetrabromobisphenol A (TBBPA) has reproductive developmental toxicity remains controversial. Here, we evaluated the effects of postnatal TBBPA exposure of dams (before weaning) and pups through drinking water (15, 150, 1500ng/mL) on testis development in mice. On postnatal day (PND) 56, we found that TBBPA exerted little effects on testis weight, anogenital distance, sperm parameters, and the serum testosterone level, but resulted in dose-dependent reductions in the seminiferous tubule area coupled with decreased Sertoli cells and spermatogonia and the number of stage VII-VIII seminiferous tubules, and cytoskeleton damage in Sertoli cells, along with down-regulated expression of marker genes for Sertoli cells, spermatogonia and spermatocyte. Further study revealed that the reduced tubule area coupled decreased Sertoli cell and germ cell numbers and marker gene expression also occurred in TBBPA-treated testes on PND 7, along with reduced cell proliferation and disordered arrangement of Sertoli cell nuclei. On PND 15, most of these testicular alterations were still observed in TBBPA-treated males, and cytoskeleton damage in Sertoli cells became observable. All observations convincingly demonstrate that postnatal exposure to TBBPA disturbed testis development in early life and ultimately caused adverse outcomes in adult testes, and that cell proliferation inhibition, the reduction in the seminiferous tubule area coupled decreased Sertoli cell and germ cell numbers and marker gene expression, and cytoskeleton damage in Sertoli cells, are early events contributing to adverse outcomes in adult testes. Our study improves the understanding of reproductive developmental toxicity of TBBPA, highlighting its risk for human health.

Estimation of the Regenerative Potential of para-Tyrosol in the Model of Testicular Insufficiency Caused by Damage to Spermatogonial Stem Cells.

The regenerative properties of p-tyrosol were investigated in a model of testicular insufficiency caused by a toxic effect on spermatogonial stem cells (single administration of paclitaxel in the maximum tolerable dose). Against the background of p-tyrosol administration, we observed an increase in the number of normal spermatogonia and Sertoli cells, stimulation of spermatogenesis, and renewal of the spermatogenic tissue. The treatment with p-tyrosol also led to a decrease in DNA damage in cells of the testicular tissue. These changes were accompanied by a decrease in the level of free radicals, an increase in antioxidant protection, and normalization of the redox potential.

Cypermethrin inhibits Leydig cell development and function in pubertal rats.

Cypermethrin is a broad-spectrum pyrethroid insecticide that is widely used. It may induce adverse endocrine-disrupting effects on the male reproductive system. Whether cypermethrin can disrupt Leydig cell development and function in the late puberty remains elusive. The objective of this study was to explore the effect of cypermethrin exposure to male rats on the development and function of Leydig cells in late puberty and explore the underlying mechanism. Thirty-six male Sprague-Dawley rats (age of 35 days) were gavaged with cypermethrin (0, 12.5, 25, and 50 mg/kg/day) from postnatal day 35-49. Cypermethrin significantly lowered serum testosterone level while elevating serum luteinizing hormone level at a dose of 50 mg/kg, without altering serum follicle-stimulating hormone level. Cypermethrin markedly decreased CYP11A1-positive Leydig cell number at 50 mg/kg without affecting SOX9-positive Sertoli cell number. It significantly down-regulated the expression of Leydig cell genes, Lhcgr, Star, Cyp11a1, and Cyp17a1 and their proteins, while up-regulating the expression of Sertoli cell genes, Dhh and Amh, and their proteins, at doses of 12.5-50 mg/kg. In addition, cypermethrin significantly increased malondialdehyde level while lowering the expression of Sod1 and Sod2 and their proteins at 50 mg/kg. Cypermethrin markedly induced reactive oxidative species at a concentration of 200 μM and reduced mitochondrial membrane potential at 25 μM and higher concentrations after 24 h of treatment to primary Leydig cells in vitro. In conclusion, cypermethrin inhibits the development and function of Leydig cells in male rats in late puberty.

Fertility-enhancing potential of ethanol extract of Cuscuta chinensis seeds in a rat model of unilateral cryptorchidism

&#x0D; &#x0D; &#x0D; &#x0D; Purpose: To investigate the fertility-enhancing potential of the ethanol extract of Cuscuta chinensis seeds in a rat model of unilateral cryptorchidism (ULC), and the mechanism(s) of action.&#x0D; Methods: Healthy male Sprague Dawley rats (n = 48; mean weight = 220 ± 10 g) were randomly assigned to 4 groups (12 rats/group): control, ULC, 100 mg extract/kg and 200 mg extract/kg groups. Unilateral cryptorchidism of right testis was induced via standard method using an operating microscope. Rats in the treatment groups received 100 and 200 mg of ethanol extract of Cuscuta chinensis/kg orally once a day for 60 days. Sperm count and sperm motility were determined in seminal vesicular fluid (SVF) suspension. Oxidative stress markers and histological changes in rat testis were evaluated. The levels of caspase-3 and caspase-9 in testicular tissue were assessed by enzyme-linked immunosorbent assay (ELISA), while the protein expressions of Nrf2 and heme oxygenase 1 (HO-1) were determined using Western blotting.&#x0D; Results: Body and reproductive organ weights, sperm count, sperm motility, and activities of glutathione peroxidase (GPx), catalase and superoxide dismutase (SOD) were significantly reduced in ULC group, relative to control group, but these parameters were significantly and dose-dependently increased following extract treatment (p &lt; 0.05). Malondialdehyde (MDA), 8-hydroxy-2’-deoxyguanosine (8-OHdG), caspase-3 and caspase-9 levels were significantly higher in ULC group than in control group, but they were reduced significantly and dose-dependently after extract treatment (p &lt; 0.05). Moreover, the protein expressions of Nrf2 and HO-1 were significantly downregulated in ULC group, when compared with control group, but they were significantly and dose-dependently upregulated by the extract (p &lt; 0.05). Cross sections of testicular tissues of rats in ULC group revealed narrowed and thickened seminiferous tubules (disrupted spermatogonia) characterized by increased apoptotic bodies (increased number of necrotic Sertoli and Leydig cells). However, there were few damaged or necrotic Sertoli and Leydig cells, and complete absence of thickening of seminiferous tubules in testicular tissues of rats treated with the extract.&#x0D; Conclusion: The ethanol extract of Cuscuta chinensis seeds effectively mitigates cryptorchidism in rats via mechanisms involving the regulation of Nrf2/HO-1 signaling pathway, and inhibition of apoptosis and oxidative stress. Thus, the plant extract has potentials for further development for the management of male infertility&#x0D; &#x0D; &#x0D; &#x0D;

Human INHBB Gene Variant (c.1079T&amp;gt;C:p.Met360Thr) Alters Testis Germ Cell Content, but Does Not Impact Fertility in Mice

Testicular-derived inhibin B (α/β B dimers) acts in an endocrine manner to suppress pituitary production of follicle-stimulating hormone (FSH), by blocking the actions of activins (β A/B/β A/B dimers). Previously, we identified a homozygous genetic variant (c.1079T>C:p.Met360Thr) arising from uniparental disomy of chromosome 2 in the INHBB gene (β B-subunit of inhibin B and activin B) in a man suffering from infertility (azoospermia). In this study, we aimed to test the causality of the p.Met360Thr variant in INHBB and testis function. Here, we used CRISPR/Cas9 technology to generate InhbbM364T/M364T mice, where mouse INHBB p.Met364 corresponds with human p.Met360. Surprisingly, we found that the testes of male InhbbM364T/M364T mutant mice were significantly larger compared with those of aged-matched wildtype littermates at 12 and 24 weeks of age. This was attributed to a significant increase in Sertoli cell and round spermatid number and, consequently, seminiferous tubule area in InhbbM364T/M364T males compared to wildtype males. Despite this testis phenotype, male InhbbM364T/M364T mutant mice retained normal fertility. Serum hormone analyses, however, indicated that the InhbbM364T variant resulted in reduced circulating levels of activin B but did not affect FSH production. We also examined the effect of this p.Met360Thr and an additional INHBB variant (c.314C>T: p.Thr105Met) found in another infertile man on inhibin B and activin B in vitro biosynthesis. We found that both INHBB variants resulted in a significant disruption to activin B in vitro biosynthesis. Together, this analysis supports that INHBB variants that limit activin B production have consequences for testis composition in males.

Cold plasma promotes Sertoli cell proliferation via AMPK–mTOR signaling pathway

This study investigated cold plasmas for multiple biological applications. Our previous work has found dielectric barrier discharge plasma improves chicken sperm quality. The number of Sertoli cells (SCs) decides spermatogenesis. However, whether cold plasma can regulate SC proliferation remains unclear. This study explored the effects of cold plasma on immature chicken SC proliferation and the regulation mechanism. Results showed that cold plasma exposure at 2.4 W for 30 s twice with an interval of 6 h produced (P<0.05) the maximum SC viability, cell growth, and cell cycle progression. SC proliferation-promoting effect of cold plasma treatment was regulated by increasing (P<0.05) the adenosine triphosphate production and the respiratory enzyme activity in the mitochondria. This process was potentially mediated by the adenosine monophosphate-activated protein kinase (AMPK)–mammalian target of rapamycin (mTOR) signaling pathway, which was regulated by the microRNA (miRNA) targeting regulation directly and by the intracellular reactive oxygen species homeostasis indirectly. The cold plasma treatment increased (P<0.01) the miR-7450-5p expression and led to a decreased (P<0.01) AMPKα1 level. On the other hand, miR-100-5p expression was reduced (P<0.05) and led to an increased (P<0.05) mTOR level in SCs. A single-stranded synthetic miR-7450-5p antagomir and a double-stranded synthetic miR-100-5p agomir reduced (P<0.05) the SC proliferation. However, this could be ameliorated (P<0.05) by the cold plasma treatment. Our findings suggest that appropriate cold plasma treatment provides a safe strategy to improve SC proliferation, which is beneficial to elevating male chicken reproductive capacity.

Effects of Lycium barbarum polysaccharides on the proliferation and differentiation of primary Sertoli cells in young rats

Sertoli cells (SCs) provide physical support and material supply for germ cells and participate in the formation of blood-testis barrier. The number of SCs is directly proportional to the number of germ cells. And mature SCs ensure the growth of germ cells and the production of sperm. In this study, we explored the effect and underlying mechanism of Lycium barbarum polysaccharides (LBP) on primary SCs in young rats. Primary SCs were isolated from the testis of 20-day old rats. The cells were then treated with different concentrations of LBP. Immunocytochemistry was used to detect the expression of Ki67 and the androgen receptor (AR), and western blotting was used to detect the expression of cytokeratin-18 (CK-18), AR and phosphorylated Akt (Ser473) in SCs. The number of SCs increased significantly after LBP treatment, and the 100 mg/mL. LBP group had 14% more cells than the control group. The expression of Ki67 in LBP treated groups also increased significantly. LBP inhibited the expression of cytokeratin 18 in SCs. Besides, LBP increased the expression of AR on SCs and promoted the activation of Akt at the ser473 phosphorylation site. LBP promotes the proliferation of immature SCs in young rats and also accelerates their differentiation and maturation. This seems to be associated with activation of the Akt signaling pathway via up-regulation of AR.

Distribution of extracellular matrix related proteins in normal and cryptorchid ziwuling black goat testes.

The Ziwuling black goat is an indigenously in China, their offspring are frequently affected by congenital cryptorchidism. The extracellular matrix (ECM) contains cytokines and growth factors that regulate the development of the testis, and component changes often result in pathological changes. Cryptorchidism is closely related to structural changes in ECM. In this study, the histochemical staining, immunohistochemical, immunofluorescence and Western blot combined with semi-quantitative analysis was used to describe the distribution of the important ECM components Collagen type IV (Col IV), laminin (LN)and heparan sulfate proteoglycans (HSPG) in the normal and cryptorchid testes of Ziwuling black goats. Results showed that: The histochemical staining showed that the dysplasia of seminiferous tubules and decreased number of Sertoli cells in cryptorchidism, as well as sparse collagen fiber. Meanwhile, the distribution of reticular fibers is relatively rich. Furthermore, the PAS and AB staining in the interstitial vessels and lamina propria of seminiferous tubules is weak. The immunohistochemical and immunofluorescence revealed that Col IV, LN was strongly expressed in Leydig, Sertoli cells of normal testes and moderately positive in the spermatogonia and spermatids, but HSPG was not expressed in the spermatogonia. However, cryptorchidism, the expression of Col IV, LN and HPSG in Leydig, Sertoli cells significantly decreased, as well as the expression of Col IV and LN in capillary endothelial cells, but HSPG was moderately expressed in spermatogonia. Based on these data, the underdevelopment of spermatogenic epithelium, decreased synthesis function of collagen fibers and Leydig cells develop usually in the cryptorchidism were shown to be closely related to the abnormal metabolism of Col IV and LN. The positive expressed of HSPG in the spermatogonia of cryptorchid testes is related to the compensatory development of spermatogonia.

Protective effects of Equisetum arvense methanolic extract on testicular tissue disorders in streptozotocin-induced diabetic murine model.

Diabetes in a long period can damage the testicular tissue and impair the male fertility potential. Recently, different herbal treatments have been used for the prevention of type I diabetes and its pathological effects. Methanolic extract of Equisetum arvense has anti-oxidant and hypoglycemic properties. Thus, the current study aimed to evaluate the protective effects of Equisetum arvense methanolic extract (EE) on diabetes-induced detrimental effects in mice testicular tissue. Thirty-two adult male mice were randomly divided into four groups including control-sham, diabetic (induced by streptozotocin, 50.00 mg kg-1 for five days), diabetic + EE 250 (250 mg kg-1) and diabetic + EE 500 (500 mg kg-1). After 45 days, all animals were euthanized and their testicles were dissected out and undergone histological analyses. Moreover, the serum level of testosterone was evaluated. Analyses showed that seminiferous tubules diameter, Leydig cells number per mm2 of the connective tissue, Sertoli cells number per tubule, serum level of testosterone and percentage of seminiferous tubules with positive tubular differentiation, repopulation and spermiogenesis indices were significantly decreased in the diabetic group in comparison with control-sham group. The administration of EE in test groups significantly decreased the adverse effects of diabetes (especially 500 mg kg-1). The results of this study revealed that diabetes disturbs spermatogenesis and spermiogenesis processes in mice. Meanwhile, the EE prevents diabetes-induced damages in mice testicular tissue, which may be associated with its hypoglycemic and antioxidative activities.

Serial Inhibin B Measurements in Boys with Congenital Monorchism Indicate Compensatory Testicular Hypertrophy in Early Infancy.

Congenital monorchism is considered a condition in which an initially normal testis has existed but subsequently atrophied and disappeared due to a third trimester catastrophe (presumably torsion). Since inhibin B concentrations appear related to Sertoli and germ cells number, we evaluated pre- and postoperative inhibin B of boys with congenital monorchism to determine whether the well-known hypertrophy of the contralateral testis was reflected in inhibin B concentrations. Twenty-seven boys consecutively diagnosed with congenital monorchism (median age 12 months) underwent follow-up with reproductive hormones 1 year postoperatively (median age 25 months). The results were compared with inhibin B of 225 boys with congenital nonsyndromic unilateral cryptorchidism, by converting values to multiple of the median (MoM) for age in normal boys. Ten boys (37%) had blind-ending vessels and ductus deferens (vanished testis) and the remaining (63%) had testicular remnants. At the time of diagnostic procedure, monorchid boys did not have significantly lower inhibin B (median 114, range 20-208) than unilateral cryptorchid boys (136, 47-393) (p = 0.27). During follow-up, MoM values of inhibin B increased in monorchid boys (median 0.59 to 0.98) and in unilateral cryptorchid boys (0.69 to 0.89) (both p < 0.0001). Compared with the concentration at surgery, an additional 44% monorchid boys had inhibin B MoM values higher than 1.0, whereas only additional 23% of unilateral cryptorchid boys exhibited such values (p = 0.04). Generally, inhibin B MoM values were normalized during follow-up in boys with congenital monorchism, reflecting compensatory hypertrophy within the first 2.5 years of life. The compensatory capacity to increase was better in monorchism than in unilateral cryptorchidism.

Multifaceted epigenetic regulation of porcine testicular aromatase

Testicular aromatase catalyzes the synthesis of estradiol, which contributes to regulation of porcine Sertoli cell proliferation and postpubertal maintenance of Sertoli cell numbers. Although aromatase enzymatic activity decreases with age and is persistently reprogrammed by prepubertal treatment with the aromatase inhibitor letrozole, the molecular bases for regulation have not been identified. DNA methylation was examined as a potential regulatory mechanism using DNA from Leydig cells isolated from 16-, 40-, and 68-week-old boars and from 68- week-old littermates treated with the aromatase inhibitor, letrozole. Methylation levels of individual CpG dinucleotides located in the distal untranslated exon 1 of the relevant aromatase encoding gene, CYP19A3, were quite high in Leydig cell DNA, and increased further with maturity of boar (P < 0.05), while aromatase activity and transcript abundance decreased more than two-fold. However, reduced aromatase activity following letrozole treatment was not accompanied by altered DNA methylation. Testicular expression of miR378 was altered by prepubertal treatment with letrozole. The data provide evidence for two different epigenetic mechanisms that regulate aromatase expression and enzymatic activity in the boar testis.