Thickness Of Seminiferous Epithelium Research Articles

Developmental variations of the reproductive organs of ganders from different goose breeds and the underlying mechanisms.

A deep understanding of the dynamics and mechanisms of male reproductive tract development is necessary for adoption of either genetic techniques or environmental management practices for improving fertility and hatchability in poultry. However, compared with other poultry such as chickens and ducks, less is known about the age- and breed-related changes in the reproductive tract development of domestic goose ganders exhibiting relatively poor reproductive performance as well as the regulatory mechanisms. In the present study, by taking 2 Chinese domestic goose breeds (Sichuan White goose, SW and Gang goose, GE; Anser cygnoides) and one European goose breed (Landes goose, LD; Anser anser) as the experimental objects, we comprehensive analyzed the morphological, histological, and genome-wide transcriptomic variations in their testicular and external genital development during the period from hatching to sexual maturity. Results from histomorphological analysis demonstrated that the reproductive tract of all goose breeds developed in both age- and breed-dependent manners, and the left and right testis developed asymmetrically throughout posthatch development. The tenth week posthatch was a critical developmental stage for all goose ganders, because both the testicular and external genital histomorphological parameters significantly changed before and after this period. During the first 10 wk posthatch, the weight, organ index, or size of male reproductive organs developed more rapidly in SW than in LD, and so were the testicular parenchymal-to-interstitial ratio and the external genital lymphatic lumen diameter. However, the testicular seminiferous epithelium thickness, seminiferous tubule diameter, and Leydig cell number, as well as the external genital keratinized epithelium thickness were significantly higher in LD than in SW at 10 wk of age. Through comparative transcriptomics analysis and RT-qPCR validation, several pathways related to germ and somatic cell function, organ remodeling, and energy metabolism were thought to be responsible for the developmental variations in the early testicular development between Chinese and European domestic ganders, where 10 hub genes involved in the cell cycle, RNA polymerase II-dependent transcription, and mitotic cell division pathways might play essential roles. These data shed new light on the interbreed differences in the male goose reproductive tract development and the molecular mechanisms regulating male goose testicular functions and fertility.

High levels of mitochondrial dynamics, autophagy, and apoptosis contribute to stable testicular status in hibernating Daurian ground squirrels

Daurian ground squirrels (Spermophilus dauricus) experience various stress states during winter hibernation, but the impact on testicular function remains unclear. This study focused on the effects of changes in testicular autophagy, apoptosis, and mitochondrial homeostasis signaling pathways at various stages on the testes of Daurian ground squirrels. Results indicated that: (1) During winter hibernation, there was a significant increase in seminiferous tubule diameter and seminiferous epithelium thickness compared to summer. Spermatogonia number and testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels were higher during inter-bout arousal, suggesting that the testes remained stable during hibernation. (2) An increased number of mitochondria with intact morphology were observed during hibernation, indicating that mitochondrial homeostasis may contribute to testicular stability. (3) DNA fragmentation was evident in the testes during the hibernation and inter-bout arousal stages, with the highest level of caspase3 enzyme activity detected during inter-bout arousal, together with elevated levels of Bax/Bcl-2 and Lc3 II/Lc3 I, indicating an up-regulation of apoptosis and autophagy signaling pathways during hibernation. (4) The abundance of DRP1, MFF, OPA1, and MFN2 proteins was increased, suggesting an up-regulation of mitochondrial dynamics-related pathways. Overall, testicular autophagy, apoptosis, and mitochondrial homeostasis-related signaling pathways were notably active in the extreme winter environment. The well-maintained mitochondrial morphology may favor the production of reproductive hormones and support stable testicular morphology.

Exploration of tissue fixation methods suitable for digital pathological studies of the testis

BackgroundThe way of testicular tissue fixation directly affects the correlation and structural integrity between connective tissue and seminiferous tubules, which is essential for the study of male reproductive development. This study aimed to find the optimal fixative and fixation time to produce high-quality testicular histopathological sections, and provided a suitable foundation for in-depth study of male reproductive development with digital pathology technology.MethodsTestes were removed from both sides of 25 male C57BL/6 mice. Samples were fixed in three different fixatives, 10% neutral buffered formalin (10% NBF), modified Davidson’s fluid (mDF), and Bouin’s Fluid (BF), for 8, 12, and 24 h, respectively. Hematoxylin and eosin (H&E) staining, periodic acid Schiff-hematoxylin (PAS-h) staining, and immunohistochemistry (IHC) were used to evaluate the testicle morphology, staging of mouse seminiferous tubules, and protein preservation. Aperio ScanScope CS2 panoramic scanning was used to perform quantitative analyses.ResultsH&E staining showed 10% NBF resulted in an approximately 15–17% reduction in the thickness of seminiferous epithelium. BF and mDF provided excellent results when staining acrosomes with PAS-h. IHC staining of synaptonemal complexes 3 (Sycp3) was superior in mDF compared to BF-fixed samples. Fixation in mDF and BF improved testis tissue morphology compared to 10% NBF.ConclusionsQuantitative analysis showed that BF exhibited a very low IHC staining efficiency and revealed that mouse testes fixed for 12 h with mDF, exhibited morphological details, excellent efficiency of PAS-h staining for seminiferous tubule staging, and IHC results. In addition, the morphological damage of testis was prolonged with the duration of fixation time.

Dietary supplementation with barley sprouts and d-aspartic acid improves reproductive hormone concentrations, testicular histology, antioxidant status, and mRNA expressions of apoptosis-related genes in aged broiler breeder roosters

The objective was to determine effects of dietary supplementation of barley sprouts (BS) and/or d-aspartic acid (DA) on the reproductive potential of aged broiler roosters. Birds (n = 32, 50 wk old) were randomly allocated to receive dietary supplements of BS powder (2 % of basal diet), and DA (200 mg/kg BW), both, or neither, for 12 wk. Roosters were housed individually, with 14-h light/10-h dark, ad libitum feed and water, and euthanized after 12 wk. Mean (±SEM) total phenolic compounds and IC50 in methanol extract of sprouted barley were 302.8 ± 10.9 mg GAE/g and 600.8 ± 50.5 mg TE per 100 g, respectively. In weekly semen collections, sperm total and progressive motility, plasma membrane integrity, sperm concentration, and sperm production were higher (P < 0.05) in both the DA + BS and BS groups compared to the Control, but sperm abnormalities were unaffected. Feeding DA increased right, left, and combined testicular weights (P < 0.05, P < 0.05, and P < 0.01, respectively) and, the testicular index (P = 0.01). Feeding BS increased seminiferous tubule diameter (P < 0.01), whereas BS + DA increased seminiferous epithelium thickness (P < 0.01). There were more spermatogonia (P < 0.01) and Leydig cells (P < 0.05) in BS-fed roosters but Sertoli cells were highest in BS + DA (P < 0.01). Serum MDA concentrations were lowest in BS (P < 0.01), whereas serum testosterone and LH were highest in DA (P < 0.05) and BS + DA (P < 0.01), respectively. Feeding BS reduced serum total cholesterol (P < 0.05) and increased serum HDL-cholesterol (P < 0.01), with decreases in serum LDL (P < 0.01) and the LDL/HDL ratio (P < 0.01) for BS + DA compared to Control. Relative expression of glutathione peroxidase mRNA was increased by BS (P < 0.01) or DA (P < 0.05), whereas relative mRNA expression of SOD was highest in BA (P < 0.01). Control roosters were highest for both BAX (P < 0.01) and the relative expression of the BAX/BCL-2 ratio (P < 0.01), whereas BS + DA increased BCL-2 (P < 0.05). In conclusion, feeding BS, and/or DA significantly improved reproductive potential in aged broiler roosters.

(088) The Essential Role of O-GlcNAcylation in the Testis Aging

Abstract Introduction With the delay of reproductive age, reproductive disorders related to advanced age have gradually attracted widespread attention. Studies have shown that the decline in male fertility due to aging is manifested as a decrease in semen parameters and spermatogenetic cells histologically. O-GlcNAc (O-acetylglucosamine) glycosylation is a post-translational modification that exists widely in cells. The level of UDP-GlcNAc, which is the substrate of O-GlcNAcylation, changes rapidly with various nutrients and external factors. Therefore, O-GlcNAcylation can respond to glucose, amino acid, fatty acid, and nucleoside metabolism, and is considered to be an important sensor of cellular metabolism. The previous study of our project found that the level of O-GlcNAc modification in aged testis tissue was significantly increased, so this study aimed to explore the specific mechanism of O-GlcNAc modification involved in testicular aging. Objective 1.Explore the modification pattern of O-GlcNAcylation in testis 2.Mimic the high O-GlcNAcylation state of aging mice in young mice, to study the effect of elevated high O-GlcNAcylation level on testis. 3.Reveal the role of key proteins modified by O-GlcNAc in testicular aging Methods Mice were injected intraperitoneally with OGA inhibitor Thiamet-G for 35 days to increase the level of O-GlcNAc to simulate aging. CASA system was used to detect semen parameters, and chromosome spreading technology was used to study mouse testis meiosis. Using protein profiling technology, the differentially modified O-GlcNAc proteins in aged testis and young testis were screened, and point mutations were made at their O-GlcNAc sites to study the specific mechanism of key O-GlcNAc proteins involved in testis aging. Results The O-GlcNAc modification level in the testis of aged mice was significantly increased. After intraperitoneally injecting Thiamet-G into mice for 35 days, the level of O-GlcNAcylation in the testis tissue was significantly increased. High level of O-GlcNAcylation in testis damaged the sperm parameter, decreased the thickness of seminiferous epithelium and increased apoptosis in testis. Chromosome spreading experiments showed that, γH2Ax, which is the marker of DNA double strand breaks, stained unusually at autosomes in pachytene spermatocytes. The elevation of γH2Ax affected the transition of pachytene spermatocytes to diplotene. Importantly, we found that the O-GlcNAcylation of Hyou1 protein was significantly increased in the testis of aged mice. O-GlcNAcylation of Hyou1 promotes the degradation of Hyou1. An O-GlcNAc site of Hyou1 was identified at S634 and mutation of this site decreased the O-GlcNAcylation, increased the protein stability and exerts protective roles in aging testis. Conclusions In the testis of aged mice, the level of O-GlcNAcylation is increased, which leads to the arrest of spermatocyte meiosis by affecting the DNA damage repair pathway. Hyou1 is the key O-GlcNAc protein in testicular aging and mutation of this site exerts protective roles. This study revealed for the first time the specific mechanism of O-GlcNAc involved in testicular aging, and provided a new therapeutic target for clinically delaying testicular aging and treating male infertility. Disclosure No

Bisphenol A exposure induces testicular oxidative damage via FTO/m6A/Nrf2 axis during postnatal development in mice.

Bisphenol A (BPA), a commonly used plasticizer in the production of polycarbonate plastics and epoxy resins, has been shown to induce male reproductive toxicity. However, the effects of BPA exposure on early testicular development have not been thoroughly studied, and the underlying mechanism is yet to be elucidated. In the current study, neonatal male mice were exposed to BPA at 0, 0.1, and 5mg/kg, respectively, by daily subcutaneous injection during postnatal day (PND) 1-35 to explore its effects on testicular development at PND 36 (the end of the first round of spermatogenesis). Morphological analyses showed that BPA exposure significantly induced apoptosis of testicular cells (p < 0.01 and p < 0.001) and reduced the thickness of seminiferous epithelium (p < 0.01). In addition, BPA exposure significantly decreased the total antioxidant capacity of testes and levels of transcription factor Nrf2 as well as its downstream antioxidant molecules of NQO1 and GPx-1 (p < 0.05 and p < 0.01). Furthermore, global m6A modifications of mRNAs were upregulated accompanied by declined m6A demethylase (FTO) in the testes of BPA groups (p < 0.05 and p < 0.01). MeRIP-quantitative real-time polymerase chain reaction (qPCR) demonstrated that BPA exposure markedly increased the m6A modification of Nrf2 mRNA (p < 0.05 and p < 0.01). These findings suggest that upregulation of m6A induced by inhibited FTO may be involved in BPA-induced testicular oxidative stress and developmental injury during postnatal development, which provides a new idea to reveal the mechanism underlying BPA interfering with testicular development.

Selenized glucose improves rat semen quality by improving the gut microbiota and serum metabolome.

Selenium (Se), a well-known antioxidant, is important for male fertility and sperm quality. The gut microbiota is involved in vital activities and cross-talk between reproduction and the gut axis. It is still unclear whether the gut microbiota mediates the impact of selenium on semen quality, and what the underlying mechanisms may be. A selenized glucose (SeGlu) derivative is a novel organic Se compound. After 7 days of acclimation, the Sprague-Dawley (SD) male rats (230 g, 6 weeks) were divided into three drinking groups: deionized water group (CK), SeGlu 0.15 group (0.15 mg Se per L), and SeGlu 0.4 group (0.4 mg Se per L). All animals were euthanized 30 days post-treatment. Serum and intratesticular testosterone and semen parameters were measured. Metagenomic and non-targeted metabolomic approaches were used to study the effects of SeGlu on the gut microbiota and serum metabolites of rats. In both the SeGlu 0.15 Group and the SeGlu 0.4 Group, we found a significant increase in seminiferous epithelium thickness. While the SeGlu 0.4 Group had a tendency to increase with insignificant difference, the SeGlu 0.15 Group significantly improved the sperm viability, survival rate, and seminal plasma fructose. SeGlu had no effect on intratesticular testosterone levels, or abnormal sperm counts. Measured serum testosterone levels using ELISA and LC-MS, which showed a decreasing trend. ELISA did not reveal significant differences, but LC-MS indicated a significant decrease in SeGlu 0.4 group. Meanwhile, the SeGlu 0.15 Group reduced the abundance of harmful bacteria such as Rikenella, Barnesiella, Tenacibaculum, and Aeromonas while increasing the abundance of beneficial microbiota such as Intestinimonas, Christensenella, Coprococcus, and Butyrivibrio. Linear discriminant analysis Effect Size (LEfSe) identified the SeGlu 0.15 group's feature genera as Roseburia, Clostridium, Ruminococcus, and Eubacterium. Serum metabolites showed that the SeGlu 0.15 Group increased 5 beta-androstane-3,17-dione while decreasing estrone and 2-methoxyestradiol (2-MeOE2). In conclusion, the SeGlu 0.15 Group can significantly alter the levels of several sex hormones in serum, improve the quality of rats' sperm, and reduce harmful bacterial colonization. SeGlu 0.15 may be used as an effective dietary supplement.

The effect of dietary supplementation of coenzyme Q10 on reproductive variables of cadmium-challenged male Japanese quails (Coturnix Japonica).

In recent decades, efforts to produce more efficient poultry products have increased due to its high demand. Meanwhile, some stressors have a negative impact on poultry efficiency and reproduction. Cadmium (Cd) is a toxic heavy metal with a high potential for inducing reactive oxygen species. On the other hand, coenzyme Q10 (CoQ10), with antioxidant properties, exerts a free radical-neutralizing effect on biological systems under stressful conditions. This study aimed to determine the effect of dietary CoQ10 supplementation on reproductive variables of Cd-challenged male quails. Two hundred and sixteen 42-day-old Japanese quails with a male-to-female ratio of 1:3 were randomly divided into three experimental groups (n= 72) and fed by experimental diets from 9 to 13 weeks of age (woa). Treatments included a negative control (NC): feeding basal diet; positive control (PC): feeding basal diet and Cd administration (1mg per 100g body weight at 10 and 11 woa); and CdQ10: dietary supplementation of CoQ10 (900mg per kg diet) and Cd administration. At 10 and 13 woa, liver and testis, cloacal gland index, sera concentration of malondialdehyde (MDA) and testosterone, total antioxidant capacity (TAC), alanine aminotransferase (ALT), testicular histology, mRNA abundance of Hsp70 and fatty acid profile of testis, as well as hatchability and fertility, were measured. Liver and testis weights, cloacal gland index, serum concentration of testosterone, ALT, MDA, TAC, mRNA abundance of HSP70, hatchability, and fertility were not affected by the treatments. However, Cd administration decreased seminiferous tubule diameter and seminiferous epithelium thickness (SET) in the PC group compared to the NC group (p< 0.05). The proportion of saturated fatty acids (SFA) in testis tissue was increased, and the proportion of PUFA and n-3 to n-6 PUFA ratio was decreased in the PC group compared to the NC group (p< 0.05). In addition, CoQ10 supplementation ameliorated the effect of Cd on decreasing SFA and increasing n-3 to n-6 PUFA ratio proportions. In conclusion, Cd exerts several adverse effects on reproductive-associated variables; some, but not all, of them are mitigated by CoQ10 supplementation.

Evaluating the effects of betaine on testicular ischemia/reperfusion injury induced by torsion/detorsion in the rat.

In this research, the effects of betaine on testicular ischemia-reperfusion were evaluated. Forty rats were randomly divided into 4 groups of sham, torsion/detorsion (TD), torsion/detorsion with two different dosage of betaine 200 mg/kg, and 300 mg/kg, respectively. At the end of the experiment, the testosterone concentration, sperm motility, concentration and vitality, oxidative stress biomarkers including Malondialdehyde (MDA), Glutathione peroxidase (GPx), Catalase (CAT), and total antioxidant capacity (TAC) were assessed. Moreover, histopathological parameters including seminiferous tubules diameter (STD), seminiferous epithelium thickness (SET), spermatogonia nuclei diameter (SpND), Sertoli cell nuclei diameter (StND) and miotic index were evaluated. The testosterone concentration altered during torsion/detorsion and betaine could increase slightly the testosterone concentration after 15 days. Sperm motility and vitality significantly increased in the betaine treated groups compared to the TD group on days 3 and 15. Among oxidative stress biomarkers, only CAT on day 3 and GPx on day 15 were significantly higher in the betaine groups compared to the TD group. Among histopathological parameters an increase in the STD and SET in betaine-200 and betaine-300 groups were observed on 15th day of post-surgery, compared to the TD group. These findings indicate that betaine can ameliorate testicular damages triggered by torsion/detorsion.

Effects of dietary supplementation of different sources and levels of selenium on the semen quality and reproductive performance in aged broiler breeder roosters

Fertility has a great impact on economic outcome in poultry sector. However, several physiological stressors such as aging adversely affected fertilization capacity and hatching quantity and quality. This study investigated the effect of dietary supplementation of different sources and levels of inorganic and organic selenium on the semen quality and reproductive performance of aged broiler breeder roosters. A total of thirty-six 50-wk-old Ross 308 roosters were randomly allocated to 6 groups and fed with different levels of organic and inorganic selenium. Treatments were included in the basal diet (control: CG), dietary supplementation of 0.15 (SeY0.15), 0.30 (SeY0.30), and 0.45 (SeY0.45) mg/kg selenium-enriched yeast (SeY), dietary supplementation of 0.30 mg/kg commercial organic selenium (Selemax), and dietary supplementation of 0.30 mg/kg sodium selenite (SS) as an inorganic source during 12 consecutive weeks. Ejaculated volume, semen quality attributes of the collected semen samples were evaluated every week.To assess fertility, hatchability and the hatched chick quality, the semen samples collected during last 2 wk of the trial were used to artificial insemination of hens. In order to measure seminiferous tube diameter and seminiferous epithelium thickness, testicular histology was also performed at the end of the experiment. Sperm motility, plasma membrane functionality and integrity, and ejaculation volume were higher in the SeY0.45 group compared to the other groups (P < 0.05). Fertility and hatchability rate as well as seminiferous epithelium thickness and seminiferous tube diameter were improved in the SeY0.45 compared with CG, SeY0.15 and SS groups (P < 0.05). Also hatchelling quality from roosters with SeY0.45 was higher than CG and SS groups (P < 0.05). No significant differences were noted in embryonic mortality between groups (P > 0.05).In conclusion, dietary supplementation of 0.45 mg SeY improved sperm quality and reproductive performance of aged broiler breeder roosters.

Cyanidin-3-O-Glucoside Supplement Improves Sperm Quality and Spermatogenesis in a Mice Model of Ulcerative Colitis.

Impaired fertility and low sperm quality are the global health problem with high attention. It has been noted that inflammation may impact fertility by affecting testicular spermatogenesis. Cyanidin-3-O-glucoside is a natural functional pigment with various health benefits. Nevertheless, studies on the mechanism by which C3G protects male reproduction in mice with ulcerative colitis remain scarce. The purpose of this study is to illustrate the potential mechanism of C3G for improving impaired fertility caused by colitis. A DSS-induced colitis model was applied to assess the effects of sperm quality with colitis and the health benefit role of C3G. Results indicated that C3G-treated mice exhibited higher body weight, longer colon length, less crypt damage and focal inflammation infiltration. Being consistent with that, low sperm count, low testis weight, high inflammation levels and abnormal thickness of seminiferous epithelium also observed in the DSS group were significantly recovered upon C3G treatment. These findings suggested that colitis has a close link to impaired fertility. Further analysis found that C3G could significantly suppress the inflammatory mediators in serum. Results conjointly indicated that C3G might improve the impaired fertility of mice with colitis by inhibiting inflammatory cytokines through the blood–testis barrier. C3G could be a promising daily supplement for ameliorating impaired fertility caused by colitis.

Effect of orally administrated letrozole on reproduction performance and gene expression of FOXJ1, LPR2 and PVRL3 in reproductive tract in aged roosters

letrozole is an aromatase inhibitor that stops the production of estrogen through interrupting the entrance of hormone androgen into a small amount of estrogen. Therefore, the current study was developed to estimate orally administrated Letrozole on the reproductive performance and relative abundance of Foxj1, PVRL3, and LPR2 mRNA in aged roosters. Fifty-five-week old ROSS 308 breeder roosters (n = 18) were orally treated using letrozole. Primarily, the body weight of the animals was recorded, and they were randomly classified into three groups (n = 6 birds/group) receiving different doses of Letrozole, including 0, 0.015, and 0.03 mg/kg body weight/day for three weeks. At the end of the trial, seminal traits, plasma, testicular hormone levels (testosterone, estradiol, and FSH), histopathological studies, in vitro fertility, and relative abundance of testis PVRL3, epidydimal Foxj1, and LPR2 mRNA were evaluated. Based on the results, the sperm quality variables were statistically higher in the 0.03 group compared to the controls. Greater histologic parameters, such as diameter of seminiferous tubules, thickness of seminiferous epithelium, categorized epididymal region, and in vitro fertility rates were estimated for the treated groups(p < 0.001). Plasma and testicular testosterone, estradiol concentrations, and plasma FSH levels were significantly influenced by letrozll treatment (p < 0.001). Relative mRNA transcript abundance increased for PVRL3 and decreased for Foxj1 and LPR2 in treated groups. Overall, aromatase inhibitors can enhance the reproductive performance of aged commercial broiler breeder roosters. However, it can impact endocytosis and ciliogenesis events via reducing estradiol.

Pulse-wave Doppler pulsatility and resistive indexes of the testicular artery increase in canine testis with abnormal spermatogenesis

Pulse-wave Doppler (PWD) ultrasonography of the testicular artery was recently used to compare parameters in fertile and infertile males, or males with normal and abnormal semen quality, while no studies verified the concomitant histological feature of the testis. The experimental hypothesis of the present study was that the blood flow parameters measured by PWD at different levels of the testicular artery reflect sperm attributes evaluated on spermatozoa collected from the tail of the epididymis and testicular histological characteristics. Pulse-wave Doppler parameters were recorded on both testes from 16 dogs of unknown fertility. After routine castration, epididymal spermatozoa were collected and evaluated for sperm total and progressive motility, membrane integrity, and sperm morphology. Testes were submitted to histological and immunohistochemical evaluations to quantify microscopic attributes and connective tissue and vascular area extent, respectively. The findings showed that the increased PWD indexes were suggestive of testicular histological alteration since negative correlations were found between pulsatility index and resistive index, measured in the marginal testicular artery, and histological attributes such as the seminiferous epithelial thickness (R = −0.528, P = 0.016 and R = −0.554, P = 0.005, respectively) and the tubular area/total area ratio (R = −0.494, P = 0.024 and R = −0.563, P = 0.004, respectively). Some histological characteristics, such as tubular diameter, seminiferous epithelium thickness, and the tubular area/total area ratio of the testis are correlated with the percentage of primary abnormalities (R = −0.874, P = 0.001, and R = −0.847, P = 0.001, and R = - 0.865, P = 0.001, respectively). In testes with increased sperm primary abnormalities, higher pulsatility index was found compared with testes with normal spermatogenesis in supratesticular artery (1.07 ± 0.06 vs 0.82 ± 0.23, P < 0.05), marginal artery (0.73 ± 0.04 vs 0.61 ± 0.07, P < 0.05), and intratesticular artery (0.67 ± 0.03 vs 0.43 ± 0.1, P < 0.05). Similarly, the resistive index was found significantly higher in the same vessel tracts in testes with abnormal and normal spermatogenesis (0.62 ± 0.02, 0.53 ± 0.02, and 0.46 ± 0.02 vs 0.51 ± 0.06, 0.44 ± 0.04, and 0.32 ± 0.06, respectively; P < 0.05). Our data demonstrated for the first time the relationship between PWD indexes and the contextual testicular architecture, suggesting that the pulsatility and resistive indexes could be an effective tool in andrology. In light of different histological alterations in the testis, caution is warranted in the definition of thresholds for normal and abnormal testicular blood flow indexes.

Effect of Alcoholic Extract of Garden Rocket Eruca sativa Mill. Seed on Some Reproductive Traits and Carcasses Quality of Aged Local Duck Males

This study aimed to investigate the impacts of Eruca sativa seed extract on some reproductive indicators and carcasses quality for aged local duck males. Forty-five aged local ducks males with the average live-weight of (2000±5g) and two years old were randomly distributed into three treatments (15 for each). After dissolution in corn oil, T2 and T3 were supplemented with 400 and 800 mg alcoholic extract of Eruca sativa seed respectively.T1 has only given corn oil (control). Weekly, a dosage of 2 ml was twice given to the treatments by syringe. The current results showed significant (P&lt;0.05) increases in the relative weight, length, width and volume of testes in males of second and third treatments compared to the first treatment (control). Besides, the second and third treatments recorded significant (P&lt;0.05) improvements in the diameter of seminiferous tubules, seminiferous epithelium thickness (µm) of testes, the Luteinizing hormone (LH), Follicle-stimulating hormone (FSH) and the level of testosterone hormone compared to the control. Chemical and physical tests of the breast meat showed no significant among all treatments in the ratio of free fatty acids, pH and water tolerability. Regarding the peroxide number, second treatment recorded the lowest significant (P&lt;0.05) compared to other treatments. It concluded that alcoholic extract of Eruca sativa seed could enhance reproductive characteristics and significantly improve sex hormones in aged local duck males and effect on the number of peroxide in its breast meat.

Effect of Platelet-Rich Plasma (PRP) on testicular damage in streptozotocin-induced diabetic rats

Background: Diabetes mellitus (DM) is a condition which could lead to induce testicular damage due to oxidative stress by hyperglycemic stress. Several therapies have been elucidated to reduce the testicular damage; one of them is Platelet-Rich Plasma (PRP). PRP has known growth factor properties which may serve potential effects in the tissue regeneration. This study aims to determine the role of PRP on testes in streptozotocin-induced diabetic rats by evaluating its impact on testosterone serum level.Methods: A randomized clinical trial with control group design has been conducted among 20 rats in this study. The sample of rats was divided into four groups such as control, DM (streptozotocin-induced), DM-Ins (treated with insulin daily), and DM-Ins-PRP (treated with insulin daily and PRP thrice in 2 weeks). Six weeks later, the testosterone level was measured by ELISA, and the right testis was collected for histopathology findings. Data were analyzed using SPSS version 25 for Windows. Results: The testes weight in the control group was 1.61±0.37 grams, followed by DM group (0.86±0.22 grams), DM-Ins group (1.15±0.25 grams), and DM-Ins-PRP group (1.09±0.36 grams). DM reduced testicular weight, seminiferous tubules diameter, the thickness of the epithelium and testosterone serum level significantly (p&lt;0.05) compared with control. PRP administration increases the thickness of seminiferous epithelium (p&lt;0.05) significantly. Besides, it also increased the testicular weight, tubules diameter, lumen diameter, and testosterone serum level, but not significant (p&gt;0.05).Conclusion: The administration of PRP showed a significant role in repairing testicular damage among diabetic rats.

Negative effect of feeding with high energy diets on testes and metabolic blood parameters of male Japanese quails, and positive role of milk thistle seed

In this study, it was aimed to investigate the changes in testicular tissue and cell count, testicular oxidative stress and some metabolic blood parameters of male broiler Japanese quails fed with high energy diet and milk thistle (Silybum marianum) seed. One hundred-twenty male 15-day-old Japanese quail chicks were divided into 4 equal groups with 30 each. The applications in each group were repeated 3 times with 10 animals each. Control group was fed with basal diet. Milk thistle seed group was fed with diet including 1% milk thistle seed. High energy diet group was fed with high energy diet including 10% corn syrup. High energy diet + milk thistle seed group was fed with high energy diet including 10% corn syrup along with 1% milk thistle seed. The feeding period in all groups was 35 days. When the quails reached 50 days old, a total of 48, 12 from each group (4 from each replication) were euthanized and blood samples and testes were collected. Compared with the control group, significant increases in body weight, serum cholesterol and glucose level, aspartate aminotransferase activity and testicular malondialdehyde level; however, significant decreases in serum testosterone level, testicular glutathione peroxidase activity, counts of round and elongated spermatid and sperm as well as histopathologically, significant decreases in seminiferous tubular diameter and seminiferous epithelium thickness, and marked disorganization in germinal cells were determined in quails fed with high energy diet. It was observed that almost all of the disturbances in testicular tissue, cell number, oxidant-antioxidant balance and metabolic blood parameters caused by feeding with high energy diet were significantly prevented by supplementation of milk thistle seed to the diet with high energy. On the other hand, alone milk thistle seed and high energy diet + milk thistle seed administrations decreased body weight in comparison to control and high energy diet groups. As a result, feeding with high energy diet causes disturbances in testes of male quails by affecting liver metabolic functions and testicular oxidant-antioxidant balance, but milk thistle seed addition to diet plays a protective role.

EFFECT OF TRICLOSAN ON TESTEICULAR FUNCTIONS: AN EXPERIMENTAL AND AN OBSERVATIONAL STUDIES

The aim was to study associations between triclosan concentration in seminal fluid or testicular tissue and germinal and endocrine functions of the testes in men. Methods. Atogether, 53 samples of seminal fluid in men with normospermia and pathozoospermia were studied. Concentration of triclosan was determined in the seminal fluid by gas chromatography with mass spectrometry (GC-MS). Spermological assessment was conducted according to the recommendations of the World Health Organization (2010) with an assessment of the sperm DNA Fragmentation Index. In addition, the concentration of total plasma testosterone was estimated. The experimental part of the work was performed on white mature CD Wistar male rats (n = 20). For two months, the rats in the experimental group received 200 mg/kg of triclosan (Sigma-Aldrich, USA) daily with food. Functional and morphometric techniques were used to evaluate the generative and endocrine function of the testes. The concentration of triclosan in the testes was measured by the GC-MS method. Results. In 84.9 % of ejaculate samples, triclosan was found with a median concentration of 0.13 (0.05-0.22) ng/ml. Men with normospermia and pathozoospermia had statistically significantly differentd concentrations of triclosan in seminal fluid (p = 0.045) and sperm DNA Fragmentation Index (p = 0.004). In the experimental part of the study, rats receiving triclosan had significantly lower testicular mass (p = 0.001), smaller diameter of convoluted seminiferous tubule and lower thickness of seminiferous epithelium (p

Oxidative stress, inflammatory reactions and apoptosis mediated the negative effect of chronic stress induced by maternal separation on the reproductive system in male mice

Exposure to severe and long-lasting stressors during early postnatal life negatively affects development of the brain and associated biological networks. Maternal separation (MS) is a valid stressful experience in early life that adversely affects neurobiological circuits. In the present study, we aimed to evaluate the effects of MS on sperm quality and histology of the testis in adult male mice. In this study, male mice were subjected to MS during post-natal days (PND) 2–14. Sperm parameters, histological alterations in the testicular tissue, ROS production (using DCFH-DA assay), gene expression of TLR4, NLRP3, TNFα, BAX, ASC, caspase-1 and BCL-2 (using RT-PCR), protein levels of caspase-3 and caspase-8 (using western blotting), and protein levels of IL-1β, IL-18, GPx and ATP (using ELISA) as well as protein expression of caspase-1 and NLRP3 (using immunocytochemistry) were evaluated. Findings showed that MS decreased count, morphology and viability of spermatozoa. MS decreased the diameter of seminiferous tubules and decreased the thickness of seminiferous epithelium. Furthermore, MS increased the level of ROS production and decreased the concentrations of GPx and ATP. MS led to increased expression of TLR4, NlRP3, TNFα, caspase-1, ASC, IL-1β and IL-18. In addition, MS induced apoptosis as evidenced by increased BAX, caspase-3 and caspase-8 as well as decreased BCL-2 expression. We concluded that early life stress induced by MS has detrimental effects on sperm parameters and testicular tissue. Our results suggest that these effects are mediated by activation of ROS production, and alterations in mitochondrial function, inflammatory processes and apoptosis pathways.

Use of supplemental dietary coenzyme Q10 to improve testicular function and fertilization capacity in aged broiler breeder roosters

In numerous studies it has been suggested that targeting mitochondria with specific compounds could efficiently inhibit various conditions associated with oxidative stress. The treatment of aged roosters with compounds such as coenzyme Q10 (CoQ10), may improve their reproductive performance by providing protection from oxidative stress. Therefore, this study was performed to assess the effect of supplemental dietary CoQ10 on the testicular function and fertility of aged broiler breeder roosters. A total of 36 roosters)47 weeks of age) were randomly divided into dietary treatments containing either 0, 300 or 600 mg CoQ10/kg diet. Three birds were allocated to each of four replicate groups in each dietary treatment. Between 47 and 54 weeks of age, ejaculates were obtained weekly from the three roosters in each replicate group. Samples in a replicate were pooled and analyzed as a single sample. Between 51 and 54 weeks of age, seminal plasma total antioxidant capacity (TAC), alanine amino transferase (ALAT) and aspartate amino transferase (ASAT) levels were assessed. Fertility, hatchability, and sperm penetration (SP) rates were likewise evaluated. Seminal volume, sperm concentration, sperm plasma membrane functionality, sperm plasma membrane integrity, seminiferous tubule diameter and seminiferous epithelium thickness exhibited quadratic increases in response to increasing levels of dietary CoQ10. Respectively, the 429.19, 433.33, 464.50, 613.50, 392.78 and 447.99 mg/kg dietary concentrations of CoQ10 provided the best results for each of the aforementioned variables. Also, other seminal traits, as well as testosterone concentration, fertility, and SP rates, displayed linear increases in response to the increasing levels of CoQ10. Dietary supplementation of CoQ10 linearly decreased seminal plasma ALAT and ASAT and linearly increased seminal plasma TAC. In conclusion, CoQ10 supplementation in the diet (a minimum of 300 mg CoQ10/kg diet) has the potential to improve the reproductive performance of aged broiler breeder roosters.

Immunolocalization of RFamide-related peptide 3 in a desert rodent Gerbillus tarabuli during seminiferous epithelium cycle.

Gerbillus tarabuli is a nocturnal seasonal breeder desert rodent with a main breeding season in spring and summer, and sexual quiescence in winter. This species is an interesting model for studying testis function in rodents. Therefore, the present study was performed firstly to investigate the stages of seminiferous epithelium cycle of Gerbillus tarabuli with a histological, morphometric and statistical study. And secondly to investigate the expression and possible variations in cellular distribution of RFamide-related peptide-3 (RFRP-3) - the mammalian ortholog of avian gonadotropin-inhibitory hormone (GnIH) - during seminiferous epithelium cycle using immunohistochimestry. Our results showed for the first time that the seminiferous epithelium cycle in Gerbillus tarabuli comprises 14 well-defined stages according to the tubular morphology method. The seminiferous epithelium thickness showed a significant difference during the epithelium cycle, thus it was the only morphometric classification criterion of seminiferous epithelium cycle in Gerbillus tarabuli. The immunohistochemical study reveals, for the first time, the presence of RFRP-3 in Gerbillus tarabuli testes, in both testicular compartments: the tubular and the interstitial. RFRP-3 is expressed differently according to the seminiferous epithelium cycle, RFRP-3 seemed to be more expressed at the stages V-VII and XIII. RFRP-3 was detected in Sertoli cells (≈12%), spermatocytes I (≈19%), round and elongated spermatids (≈13%), and with a more important signal in Leydig cells (26.87%±0.07). These results indicated the importance of RFRP-3 in testicular function in Gerbillus tarabuli; its expression at the interstitial and germinal levels argues in favor of an involvement in androgens synthesis and in spermatogenesis specifically in meiosis and spermiogenesis. This action seems primordial from stages V-VII and XIII. Also, the study of the seminiferous epithelium cycle will enrich the histological identity of the species.