Sperm Quantity Research Articles

CRISPR/Cas9-mediated Serine protease 2 disruption induces male sterility in Spodoptera litura.

Male fertility is essential for reproduction and population growth in animals. Many factors affect male fertility, such as courtship behavior, sperm quantity, and sperm motility, among others. Seminal Fluid Proteins (SFPs) are vital components of seminal fluid in the male ejaculate, which affect male fertility, sperm activation, and female ovulation. However, the knowledge of SFPs is insufficient; the function of many SFPs remains unknown, and most described functions were mainly characterized in Drosophila or other laboratory models. Here, we focus on the Serine protease 2 (Ser2) gene in the lepidopteran pest Spodoptera litura. The Ser2 gene was specifically expressed in male adults. Disruption of the Ser2 gene mediated by CRISPR/Cas9 induced male sterility but females remained fertile. PCR-based detection of the next-generation mutants showed that male sterility was stably inherited. The qRT-PCR analysis of SlSer2 mutants showed that motor protein family genes and structural protein family genes were down-regulated, while protein modification family genes were up-regulated, suggesting that SlSer2 may be involved in sperm movement and activity. These results demonstrate that Ser2 is an important component of SFPs in seminal fluid and was identified for a useful sterile gene for pest control that may lead to new control strategies for lepidopteran insect pests such as S. litura.

Ayurvedic management of male Vandhyatwa with special reference to male infertility

Infertility is the not getting pregnant, despite of having carefully timed, unprotected sex for one year.Various environmental and biological factors affect the sperm count in males. Oligozoospermia means low sperm count and Azoospermia means complete absence of sperms is one of the main cause of male infertility and term can be correlate with KsheenaShukra andNirbeeja /Ashukra respectively. There is no satisfactory treatment in modern medicine for this condition. Asthenozoospermia means the motility of spermatozoa is lower than 50% of active motile sperm. Ayurveda has many ways to increase sperm count, sperm motility and thus increase fertility. Sperm quality, quantity and motility decreases as their age progresses. The Aging process can be slow down by using Ayurvedic Vajeekarana Chikitsa (Aphrodisiac Therapy) which rejuvenates the male reproductive system and slows down the aging process also increases the quality, quantity and motility of sperms . It has also recommended an appropriate use of plant based remedies, a proper lifestyle and nutritious diet, Panchakarma,Yoga for improving overall health and treating male infertility. Current paper aims at putting forward the causes and treatment of male infertility as described in Ayurveda.

The effects of Aroclor 1254 alone and in combination with X-rays on the male mice germ cells quantity and quality

The effects of chemical and physical environmental factors are concerned as the main reason of diminished male fertility. The aim of the study was the investigation of the effects of low doses of Aroclor 1254 or combined exposure to low doses of Aroclor 1254 and low doses of ionizing radiation on the sperm quantity and quality of male germ cells including damage to genetic material of adult male mice. Mice were exposed for 2 weeks, 3 times per week by intraperitoneal injection with Aroclor 1254 diluted in corn oil at doses of 1, 2 and 4 mg/kg bw or to whole body X-rays irradiation at doses 0.05 Gy, 0.10 Gy and 0.15 Gy or to combination of X-rays and Aroclor 1254 at following doses 0.05 Gy + 1 mg/kg bw Aroclor 1254, 0.10 Gy + 2 mg/kg bw Aroclor 1254. The samples for sperm count, motility, morphology and DNA integrity of male germ cells estimation were taken from animals just after the end of exposure and 5 weeks later. Irradiation alone deteriorated sperm count and quality. Aroclor 1254 significantly reduced the sperm motility and increased sperm abnormality and at the highest dose also induced DNA damage of gametes. The combined exposure to 0.10 Gy + 2 mg/kg bw of Aroclor 1254 showed the increase in the sperm concentration and the decrease of percentage of abnormal spermatozoa compared to results after irradiation to 0.10 Gy alone. In conclusion, the low doses of Aroclor 1254 used in this study did not significantly reduce the sperm count, but affected the sperm motility, morphology and sometimes also DNA integrity of gametes. In combination with low doses of irradiation, low doses of Aroclor 1254 may ameliorate the harmful effect of irradiation on the male gametes.

Gonad Quality of Banana Shrimp Male Broodstock Penaeus merguiensis (De Man, 1888) Fed Different Natural Diets.

A study was carried out to determine the maturation period, quality and quantity of sperms production in Banana shrimp male broodstock, Penaeus merguiensis fed different natural diets. The three different natural diets namely; squid, fish and shrimp flesh used in this study were obtained from known sources and fed to the tested shrimp in triplicate groups. Based on the results obtained, squid seem to be the most effective natural diet as it enhances the sperms maturation within 20 days. Feeding fish and shrimp flesh as diets for the Banana shrimp broodstock resulted in the observation of maturation in 22 and 24 days, respectively. Similarly, squid diet also recorded the highest (p = 0.002; p < 0.05) sperms count (58.6 to 74.5) as compared to fish diet (44.0 to 61.3) or shrimp diet (28.0 to 42.8). Also, feeding squid diet resulted in a higher percentage of live sperms with ranged between 97.75% to 98.80%. On the other hand, broodstocks fed fish and squid flesh was observed with ranges of between 96% to 97.86% and 92.54% to 96.06%, respectively. It was therefore concluded that squid diet was most effective to improve sperm quality, quantity and maturation period in male broodstock of P. merguiensis. Further study should be carried on the reproductive performance of broodstock fed with squid as diets and it effect on the performances of post larvae obtained.

P-017 Impact of yoga intervention on immunosenescence, fertility potential and quality of life in men with Rheumatoid arthritis

Abstract Study question Can a yoga intervention bring about alterations in immune aging, fertility potential and quality of life in men with Rheumatoid arthritis? Summary answer Yoga not only reduces disease severity, minimizes usage of drugs but also reduces the rate of immune aging and cellular aging. What is known already The complex mechanism of Rheumatoid arthritis with infertility in men involves interactions between endocrine, immune, and reproductive systems. Association of autoimmunity with dysregulated androgen (hypogonadism) levels may cause transient infertility in men. Furthermore, usage of disease-modifying antirheumatic drugs (DMARDs) like cyclophosphamide, methotrexate, sulphasalazine etc. may result into decreased quantity and quality of sperm, mitochondrial dysfunctions, reduced fertility potential and ultimately permanent infertility. These drugs can cross blood-testis-barrier and induce changes in sperm impairing spermatogenesis. Complementary and alternative medicine like yoga reduces seminal oxidative stress and its consequences like DNA fragmentation in sperm nuclear/mitochondrial genome. Study design, size, duration Fifty males with RA were enrolled in this 12-week prospective, open-label, single-arm exploratory study, designed to explore the impact of yoga intervention on immunosenescence, fertility potential and quality of life (QoL). Participants/materials, setting, methods The participants were evaluated for pre (day 0) and post (12th week) levels of inflammatory markers: IL-6, IL-17A, TGF-β and TNF-α. T cell sub-set populations along with their aged T cell profiles for immunosenescence as well as telomere length and telomerase activity for cellular aging were studied. Seminal reactive oxygen species (ROS), DNA fragmentation index (DFI) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels were estimated as per WHO 2010 guidelines. QoL was measured by WHOQOL-BREF Scale. Main results and the role of chance There was significant reduction in mean levels of TNF-α, IL-6, IL-17A and elevation in TGF-β after 12 weeks. A significant reduction was seen in seminal ROS levels, 8-OHdG and DFI as well. There was a significant decline in aged T cell populations after yoga. There was no change seen in the telomere length but there was an improvement in the telomerase activity. The participants showed significant improvements in QoL after 12 weeks. Limitations, reasons for caution Compliance of patients for Yoga intervention was poor, hence we enrolled large number of patients to achieve the desirable sample size. Wider implications of the findings Adoption of yoga intervention holds the key to maintaining immune homeostasis, reduction of inflammation, reduction in the rate of cellular aging, improved overall QoL, and its associated consequences associated with physical, mental, and reproductive health. Trial registration number N/A

The computational study, thermodynamic parameters, and partial inhibition of spermatogenesis exhibited by toxicophores scaffolds of an organic–inorganic hybrid complex of tributyltin (IV)

The present investigation focused on the structure and demonstration of partial inhibition of spermatogenesis in male albino rats through pentacoordinate tributyltin (IV) complex of sterically blocked Schiff base of heterocyclic β‐diketone having the general formula Bu3SnL (where ). The aid of physicochemical and spectral analysis suggested the plausible structure of this complex. The density functional theory (DFT) provides the optimized structure of the tributyltin (IV) complex. The computational study of the tributyltin (IV) complex evaluated the thermodynamic parameters, molecular orbitals, principal quantum chemical descriptors, and chemical reactivity. The experimental animals contain three groups, each consisting of nine animals. The male fertility of albino rats of groups II and III treated with tributyltin (IV) complex of sterically blocked Schiff base of heterocyclic β ‐diketone at 10 and 20 mg/kg body wt/day compared with group I during 90 days of the investigation was impaired consistently. Accordingly, the quality and quantity of sperms obtained from cauda epididymis deteriorated in both treatment groups compared with control animals. Blood testes barrier ruptured in the seminiferous tubules as a result of disordered germinal epithelium, substantial loss of germ cells, and remaining germ cell bursting, which stopped spermatogenesis at stage 13 of spermiogenesis. The findings of the study suggest that this complex partially inhibits spermatogenesis. Identifying some toxicophores scaffolds of the complex appears to be a significant contributor to the complete inhibition of spermatogenesis.

Estimating female malaria mosquito age by quantifying Y-linked genes in stored male spermatozoa

Vector control strategies are among the most effective measures to combat mosquito-borne diseases, such as malaria. These strategies work by altering the mosquito age structure through increased mortality of the older female mosquitoes that transmit pathogens. However, methods to monitor changes to mosquito age structure are currently inadequate for programmatic implementation. Female mosquitoes generally mate a single time soon after emergence and draw down spermatozoa reserves with each oviposition cycle. Here, we demonstrate that measuring spermatozoa quantity in female Anopheles mosquitoes is an effective approach to assess mosquito age. Using multiplexed qPCR targeted at male spermatozoa, we show that Y-linked genes in female mosquitoes are exclusively found in the spermatheca, the organ that houses spermatozoa, and the quantity of these gene sequences significantly declines with age. The method can accurately identify mosquitoes more than 10 days old and thus old enough to potentially transmit pathogens harbored in the salivary glands during blood feeding. Furthermore, mosquito populations that differ by 10% in daily survivorship have a high likelihood of being distinguished using modest sample sizes, making this approach scalable for assessing the efficacy of vector intervention control programs.

Maternal exposure to PM2.5 disrupting offspring spermatogenesis through induced sertoli cells apoptosis via inhibin B hypermethylation in mice

Particulate Matter 2.5 (PM2.5) disrupts endocrine functions and may negatively affect sperm quality and quantity in males; however, the long-term effects and potential mechanisms of this effect are unknown. This study aimed to investigate the epigenetic mechanism of maternal exposure to PM2.5-induced inhibin B hypermethylation in male offspring. In this experiment design, pregnant C57BL/6 mice were treated with two doses of PM2.5 (4.8 and 43.2 mg/kg bw). The membrane control group was given a sampling membrane and the control group received nothing. Following the formation of the vaginal plug, intratracheal instillation of PM2.5 was administered every three days until delivery of the pups. To assess the effect of PM2.5 in vitro, TM4 cells, a Sertoli-like cell line, was treated with different concentrations (0, 25, 50, 100 μg/mL) of PM2.5 for 24 h. The results displayed that Sperm motility, as well as the number of adult offspring, was decreased in the PM2.5 exposed group relative to the untreated controls. Increased vacuolization was observed in the Sertoli cells of mice that were exposed to PM2.5 in utero. The levels of inhibin and testosterone were reduced and the levels of LH and FSH increased in the PM2.5 groups relative to the untreated controls. In vitro, PM2.5 resulted in cell cycle inhibition as well as increased apoptosis in TM4 cells. Moreover, PM2.5-induced inhibin B hypermethylation and activation of the p21/Cleaved Caspase-3 pathway resulted in TM4 cell apoptosis that was rescued through the use of a DNA methylation inhibitor. Together, our data suggest that prenatal exposure to PM2.5 results in inhibin B hypermethylation and can activate the p21/Cleaved Caspase-3 pathway, resulting in Sertoli cell apoptosis, aberrant secretion of androgen binding protein, and decreased testosterone, thus resulting in the inhibition of spermatogenesis.

17α-estradiol does not adversely affect sperm parameters or fertility in male mice: implications for reproduction-longevity trade-offs.

17α-estradiol (17α-E2) is referred to as a nonfeminizing estrogen that was recently found to extend healthspan and lifespan in male, but not female, mice. Despite an abundance of data indicating that 17α-E2 attenuates several hallmarks of aging in male rodents, very little is known with regard to its effects on feminization and fertility. In these studies, we evaluated the effects of 17α-E2 on several markers of male reproductive health in two independent cohorts of mice. In alignment with our previous reports, chronic 17α-E2 treatment prevented gains in body mass, but did not adversely affect testes mass or seminiferous tubule morphology. We subsequently determined that chronic 17α-E2 treatment also did not alter plasma 17β-estradiol or estrone concentrations, while mildly increasing plasma testosterone levels. We also determined that chronic 17α-E2 treatment did not alter plasma follicle-stimulating hormone or luteinizing hormone concentrations, which suggests 17α-E2 treatment does not alter gonadotropin-releasing hormone neuronal function. Sperm quantity, morphology, membrane integrity, and various motility measures were also unaffected by chronic 17α-E2 treatment in our studies. Lastly, two different approaches were used to evaluate male fertility in these studies. We found that chronic 17α-E2 treatment did not diminish the ability of male mice to impregnate female mice, or to generate successfully implanted embryos in the uterus. We conclude that chronic treatment with 17α-E2 at the dose most commonly employed in aging research does not adversely affect reproductive fitness in male mice, which suggests 17α-E2 does not extend lifespan or curtail disease parameters through tradeoff effects with reproduction.

Common carp (Cyprinus carpio) sperm reduction during short-term in vitro storage at 4 °C

This study was designed to optimize a short-term storage protocol for common carp sperm at 4 °C under aerobic condition. Sperm from individual males were collected directly with or without extenders. The results demonstrated that in general, it was similar effect to collect sperm directly in extenders and keeping sperm for 0.5 h after collection without extenders. Sperm was diluted with eight selected extenders (sperm: extender = 2:1, 1:1 and 1:9) and undiluted sperm was used as a control. Sperm and seminal plasma parameters (sperm motility, velocity, membrane integrity, sperm concentration, osmolality and pH in seminal plasma) were evaluated in sperm stored on ice under aerobic conditions at 0, 2, 4, 6 and 8 days post stripping (DPS) using the computer- assisted sperm analysis system. Results showed that 1:1 and 2:1 dilution maintained higher sperm function and more sperm for a longer period. After 8 DPS, the best sperm quality and quantity was recorded in the common carp seminal plasma supplemented with 50 mM NaCl, Cejko extender (2 mM CaCl2, 1 mM MgSO4, 20 mM Tris, 110 mM NaCl, 40 mM KCl, pH 7.5 and 310 mOsm/kg) supplemented with/without 25 mM KCl/NaCl. The reduction of spermatozoa number with time during short-term storage but varied according to different dilution ratios and extenders. At 8 DPS, sperm count has dropped by 22.9 % in a dilution of 1:1 compared to 50.3 % in sperm without dilution. Extenders with diluted 1:1, especially Cejko solution, largely postponed sperm reduction, 21.3 % compared to 55.5 % for sperm stored without extenders.

Apigeninidin-enriched Sorghum bicolor (L. Moench) extracts alleviate Aflatoxin B1-induced dysregulation of male rat hypothalamic-reproductive axis.

We examined the protective effect of the apigeninidin (API)-enriched fraction from Sorghum bicolor sheaths extracts (SBE-05, SBE-06, and SBE-07) against aflatoxin B1 (AFB1)-induced dysregulation of male rat's reproductive system that may trigger infertility. Male rats (160 ± 12 g) were treated with AFB1 (50 µg/kg) along with 5 or 10 mg/kg of SBE-05, SBE-06, and SBE-07 for 28 days. Subsequently, we assessed the reproductive hormone-prolactin, FSH, LH, testosterone levels, and testicular function enzymes. Moreover, we examined rats' testes, epididymis, and hypothalamus for oxidative and inflammatory stress biomarkers, caspase-9 activity and tissues pathology. We observed that comparative to AFB1 alone treated rats, API co-treatment significantly (p < 0.05) abated the AFB1-mediated decrease in prolactin and antioxidant defenses and lessened lipid peroxidation (LPO) and reactive oxygen and nitrogen species levels in the examined organs-testes, epididymis, and hypothalamus. API abated AFB1-induced hormone decreases-testosterone, FSH, and LH; and caused improvement in sperm quantity and quality. API lessened AFB1-mediated increase in pro-inflammatory cytokine, increased interleukin-10 level, an anti-inflammatory cytokine and reduced caspase-9 activities. In addition, API reduced alterations in the examined tissue histology. Our findings suggest that S. bicolor API-enrich extracts have active antioxidative, antiapoptotic, and anti-inflammatory activities, which can protect against AFB1-induced dysfunction of the hypothalamic-pituitary-gonadal axis.

Shenjing Guben Wan promotes sperm development by increasing the activity of seminiferous epithelium Sertoli cells.

BackgroundInfertility is an important social problem. Asthenozoospermia (AZS) is a common pathological cause of male infertility, but its pathogenesis is unclear. Shenjing Guben Wan (SJGBW), a traditional Chinese medicine, has shown remarkable effects during the clinical treatment of oligozoospermia or AZS.MethodsIn this study, clinical evaluations were carried out on 184 AZS patients receiving SJGBW treatment, including sperm count, sperm quality, and pregnancy rate. Also, ornidazole was used to build an AZS mouse model, and SJGBW treatment was administered. The sperm quantity and fertility of mice in different groups were evaluated; a cholecystokinin octapeptide-8 (CCK-8) experiment was carried out to test the activity of seminiferous epithelium Sertoli cells, and immunohistochemistry and the Terminal Deoxynucleotidyl Transferase-mediated dUTP Nick-End Labeling (TUNEL) method were employed to test the pathological information and expression of the Sertoli cell surface marker in the testicular tissues of mice in each group.ResultsThe sperm vitality, progressive sperm motility, and sperm morphology of patients who received SJGBW treatment were all improved (P<0.05). In the AZS group, the average sperm count, sperm vitality, pregnancy rate, and female mouse litters were all lower relative to mice in the control group. Following SJGBW treatment, the average sperm count, sperm vitality, pregnancy rate, and female mouse litters of mice in the AZS group were all significantly improved. The cytobiological experimental results showed that compared with the serum of normal male mice in the control group, the drug serum containing SJGBW could improve the cell vitality and proliferative ability of seminiferous epithelium Sertoli cells in AZS mice. Furthermore, the TUNEL results showed that the seminiferous tubule Sertoli cells and mesenchymal cells of the AZS mice exhibited the most significant apoptosis, which was alleviated following SJGBW treatment. Moreover, the levels of Sertoli cell marker, SOX9, and anti-apoptosis protein, Bcl2, in SJGBW-treated mice were both higher than that in AZS mice.ConclusionsSJGBW can promote the development and maturation of germ cells by facilitating the proliferation of Sertoli cells in AZS patients, thereby improving the fertility of these patients.

Evidence for Sperm Limitation in the Louisiana Blue Crab Spawning Stock

AbstractMale‐targeted fishing pressure has the potential to skew population sex ratios and to result in sperm limitation in females, where the quantity or quality of sperm received during mating is insufficient for the female to achieve her full lifetime reproductive potential. Blue crabs Callinectes sapidus are heavily targeted by commercial and recreational fisheries throughout much of their range, with fishing pressure biased toward males. Additionally, blue crabs are particularly susceptible to sperm limitation because mating occurs during a short window following the female’s terminal molt and stored sperm is used to fertilize all future broods. We investigated sperm quantities in recently molted and mated female blue crabs in southeastern Louisiana to assess the potential for sperm limitation in this area. Sperm quantities were an order of magnitude lower than those previously observed in other areas. Brood production modeling indicated that most of the female blue crabs in this region are heavily sperm limited and unable to achieve their lifetime spawning potential. On a population level, this sperm limitation may be partially responsible for recent declines in landings and estimates of abundance. Although insufficient data exist to assess the precise cause of this sperm limitation, these results should be incorporated into future stock assessments and management decisions for the Louisiana blue crab spawning stock.

Effects of inbreeding depression on the success of artificial reproduction in the African catfish Clarias Gariepinus (BURCHELL, 1822)

The objective of this study was to establish an effective method of artificial reproduction and larval rearing to improve the fry production of the African catfish Clarias gariepinus. Thus, a hormonal treatment using ovaprim was used to induce maturity in males and females. Two breeding trials were first conducted on captive populations by crossing a male and female Senegalese strain (♂ss/♀ss), and a male and female Beninese strain (♂bs/♀bs). A third reproduction test was carried out by crossing ♀ss/♂bs and ♂ss/♀bs but for this test, the ♀ss and ♂ss are wild breeders that were collected from the natural environment. For the first two breeding tests (♂ss/♀ss and ♂bs/♀bs, respectively), fertilized eggs either did not hatch or some hatched but the fry did not survive. Although the number of broodstock used in these first two breeding tests is small, this lack of hatching and poor larval survival may reflect inbreeding depression. The third breeding test was successful as females and males, respectively, produced large quantities of eggs (13g of eggs for ♀ss and 32g of eggs for ♀bs) and sufficient quantities of sperm to fertilize the eggs (approximately 12g for the Benin strain and 5g for the Senegal strain). Hatching rates of 90% and 60% were obtained for the ♀ss/♂bs and ♀bs/♂ss breeding’s, respectively, after incubation of fertilized eggs on water hyacinth (natural substrate) and pompon (artificial substrate). Comparisons of growth rates of larvae fed two different diets (combination of natural and artificial food, and artificial food alone) showed that artificial food alone was more effective for growth, especially after one month of rearing. The high larval mortalities recorded especially at the end of the experiment were mainly due to poor water quality. Thus, this study provided a better understanding of the conditions in the hatchery and larval culture systems that are critical to the success of artificial reproduction and optimal growth of C. gariepinus fry.

Improvement in some Reproductive Parameters of Male Wistar Rats Administered with Leaf Extract of Eugenia uniflora

This study was designed to evaluate the effect of hydro-methanol leaf extract of Eugenia uniflora on lead-induced reproductive toxicity in male Wistar rats. The male rats used for the study were divided into 5 groups of 6 animals each. Group I was the negative control group which received no treatment. Group II was the positive control which received 5 mg/kg BW Lead daily, while Group III received 5 mg/kg BW of Lead and 200mg/kg BW of extract., Group IV received 5 mg/kg BW of Lead and 400mg/kg BW of extract and group V received 5 mg/kg BW of Lead and 800mg/kg BW of extract. The administration was by oral gavage once daily for 30 days. At the end, animals were sacrificed under light chloroform anesthesia. Blood samples were collected into plain bottles for hormonal assay. The epidydimis was located with a small incision on the abdomen and semen sample was collected for analysis of sperm parameters. The data was analysed by applying one-way analysis of variance (ANOVA) using the SPSS version 23 software. Results of this study showed that extract of Eugenia uniflora significantly increased the level of serum testosterone and the percentages of viable sperm cells, sperm cells with normal morphology and actively motile sperm cells as well as, sperm count. The extract ameliorated lead induced reproductive toxicity. The result therefore, show that the extract improves sperm quality and quantity and protects the male reproductive function from the hazardous effect of environmental toxicants such as lead.

Bi-allelic variants in KCNU1 cause impaired acrosome reactions and male infertility.

Are there new genetic factors responsible for male infertility with normal sperm quantity and morphology? We identified the bi-allelic variants in KCNU1 and confirmed it a novel pathogenetic gene for male infertility mainly due to impaired sperm acrosome reactions (ARs). Until now, the underlying genetic determinants for male affected individuals exhibiting normal sperm quantity and morphology have been largely unknown. Potassium/calcium-activated channel subfamily U member 1 (KCNU1) is a sperm-specific potassium channel. The Kcnu1 null mutation in male mice causes infertility due to the impaired progressive motility and AR. We recruited a cohort of 126 male infertility individuals with typical asthenospermia or fertilization failure and focused on two infertile males from two consanguineous families from 2015 to 2020; whole-exome sequencing and homozygosity mapping were performed. We identified a homozygous missense variant (c.2144A>G, p.His715Arg) and a homozygous donor splice-site variant (c.1295 + 3A>C, p.Val405Glyfs*8) in KCNU1. Then, we generated a knock-in (KI) mouse model in September 2020 and have now carried out functional studies and possible treatment strategies. The affected individuals with infertility were recruited from the Shanghai Ninth Hospital affiliated to Shanghai Jiao Tong University. Genomic DNA from the affected individual was extracted from peripheral blood. Whole-exome sequencing, homozygosity mapping and in silico analyses were used to screen and identify KCNU1 variants, and the variants were confirmed by Sanger sequencing. We used C57BL/6N mouse to construct KI mouse model to mimic the reproductive phenotype in vivo. We performed functional experiments by western blotting, AR assay and immunofluorescent Staining. Finally, we performed IVF and ICSI to explore the treatment strategies. We identified a homozygous missense variant (c.2144A>G, p.His715Arg) and a homozygous donor splice-site variant (c.1295 + 3A>C, p.Val405Glyfs*8) in KCNU1 in two infertile males. We demonstrated that the splice-site variant affected normal alternative splicing of KCNU1, thus leading to the loss of function of KCNU1. Meanwhile, the missense pathogenic variant reduced the KCNU1 protein levels in sperm of both the affected individual and the KI mouse model, resulting in impaired ARs and male infertility. Intracytoplasmic sperm injection was able to rescue the deficiencies. N/A. The exact molecular mechanism of KCNU1 and pathways need to be further explore in the future. This is the first report that establishes a causal relationship between KCNU1 deficiency and male infertility, confirming the critical role of KCNU1 in human reproduction. Our findings expand our knowledge of the genes that play critical roles in the human sperm AR and provide a new genetic marker for infertility. This work was supported by the SHIPM-pi fund no. JY201801 from the Shanghai Institute of Precision Medicine, Ninth People's Hospital Shanghai Jiao Tong University School of Medicine, the National Natural Science Foundation of China (81725006, 81771649, 81822019, 81771581, 81971450, 81971382, 82001538 and 82071642). The authors declare no conflict of interest. N/A.

Automated rare sperm identification from low-magnification microscopy images of dissociated microsurgical testicular sperm extraction samples using deep learning

To develop a machine learning algorithm to detect rare human sperm in semen and microsurgical testicular sperm extraction (microTESE) samples using bright-field (BF) microscopy for nonobstructive azoospermia patients. Spermatozoa were collected from fertile men. Testis biopsies were collected from microTESE samples determined to be clinically negative for sperm. A convolutional neural network based on the U-Net architecture was trained using 35,761 BF image patches with fluorescent ground truth image pairs to segment sperm. The algorithm was validated using 7,663 image patches. The algorithm was tested using 7,663 image patches containing abundant sperm, as well as 7,985 image patches containing rare sperm. Invitro fertilization center and university laboratories. Normospermic and nonobstructive azoospermia patients. None. Precision (positive predictive value [PPV]), recall (sensitivity), and F1-score of detected sperm locations. For sperm-only samples, our algorithm achieved 91% PPV, 95.8% sensitivity, and 93.3% F1-score at ×10 magnification. For dissociated microTESE samples doped with an abundant quantity of sperm, our algorithm achieved 84.0% PPV, 72.7% sensitivity, and 77.9% F1-score. For dissociated microTESE samples doped with rare sperm, our algorithm achieved 84.4% PPV, 86.1% sensitivity, and 85.2% F1-score. Rare sperm can be detected in patients' testis biopsy samples for potential subsequent use in invitro fertilization-intracytoplasmic sperm injection. A machine learning algorithm can use BF images at ×10 magnification to accurately detect sperm locations using automated imaging.

Sperm granulomas: Predictive factors and impacts on patency post vasectomy reversal.

The objective of this study was to identify factors that predict for sperm granuloma formation and the impact of sperm granuloma presence and quantity on vasectomy reversal (VR) outcomes. A cross sectional retrospective review of prospectively collected data, on the impact of granuloma on VR outcomes from a single academic center was performed. The impact of age, obstructive interval, intraoperative vasal fluid findings, anastomosis type, body mass index, tobacco use and total motile count (TMC) was determined. A total of 1550 men underwent VR between January 2000 and August 2019. Granulomas were present unilaterally in 23.3% (n = 361) and bilaterally in 14.2% (n = 220). On univariate analysis, increasing patient age negatively correlated with a larger number of granulomas (p = .011). Granuloma presence was associated with finding intact and motile sperm from the vasal stump intraoperatively (p = .001), and vasoepididymostomy anastomosis (p < .001). However, granuloma presence (and quantity) did not correlate with obstructive interval or maximum TMC. Tobacco use and body mass index (BMI) were not associated with granuloma presence. On multivariate analysis, granuloma quantity was not associated with TMC. Obstructive interval and vasovasostomy anastomosis were associated with higher TMC, while BMI was negatively associated with TMC. In conclusion, increasing age was negatively correlated with granuloma formation. Granuloma presence was associated with more favourable intraoperative fluid findings and anastomosis type, but not post‐VR TMC, suggesting men with and without granulomas undergoing skilled microsurgery will have similar patency rates. Heavier men should be encouraged for weight loss prior to vasectomy reversal as increasing BMI was associated with lower TMC.

Association study of NR5A1 rs1110061 with infertile male in 401 Vietnamese individuals

Male infertility is a reproductive issue involving defects in the quantity and quality of sperm. Besides the exogenous elements such as harmful habits, pollution, genetic factors including disorders in a single gene, group of genes, or chromosomes, are the etiology causing infertility in men. NR5A1 is well-known as a candidate gene involved in male infertility, including 46, XY disorders of sex development (DSD), cryptorchidism, micropenis, spermatogenic failure, non-obstructive azoospermia, and oligozoospermia. This study aimed to identify the single nucleotide polymorphism (SNP) associated with male infertility in the NR5A1 gene in a Vietnamese cohort of 202 infertile men and 199 healthy controls. By directly sequencing the coding region of the NR5A1 gene in 56 cases and 21 controls, only one missense mutation c.437G&gt;C (p.Gly146Ala; rs1110061) in exon 4 was found. The variant was detected in 34 patients, of which 27 were heterozygous (GC) and 7 were homozygous (CC). In the control group, 15 individuals carried this mutation, including 10 with heterozygous (GC) and 5 with homozygous (CC). To further investigate the association of the polymorphism NR5A1 rs11100061 with male infertility disease, we performed PCR-RFLP in 202 infertile and 199 healthy men to assess the genotypes and alleles. The results indicated that the distribution of the genotypes of the polymorphism was in accordance with Hardy-Weinberg equilibrium (p-values &gt; 0.05). However, no association was detected between the polymorphism and male infertility (p-values &gt; 0.05).

Putative Degradation of Non-Stored Sperm in the Female Reproductive Tract of the Dengue Vector Mosquito Aedes aegypti

In insect vectors of disease, male and female molecules that mediate reproductive processes are promising targets to suppress fertility of these populations. One process, the storage of sperm in the female reproductive tract, is essential for optimal fertility in all organisms examined to date. In the dengue vector mosquito Aedes aegypti, female sperm storage has not been fully characterized, a requirement to identify sex-specific molecules that mediate this process. Aedes aegypti males deposit the ejaculate into the bursa of the female reproductive tract, and sperm enter the spermathecae—the long-term storage sites—quickly after insemination. However, the proportion of sperm received during mating that are stored in the spermathecae is unclear, and the fate of non-stored sperm unknown. We quantified sperm storage in two Ae. aegypti strains, mated in all combinations, and in two contexts (mass mated and when mating was observed) at 1-, 3- and 5-days post-mating. Sperm quantity in the spermathecae was similar at all timepoints; most females stored ~400 sperm on average. Sperm that did not enter the spermathecae remained in the bursa, where they declined in number and became more fragile to mechanical manipulation at each timepoint. Further, sperm viability in the bursa fell from 91.6% shortly after mating to 12.2% 24 h later. One day after insemination, ~50% of sperm detected in the female reproductive tract was stored in the spermathecae. When we quantified sperm storage in females mated to males that transferred reduced ejaculate quantities (but still able to induce optimal fertility in their mates), sperm detected in the spermathecae similarly declined; females stored ~50% of the sperm received even as sperm quantities transferred at mating declined. Our results suggest that sperm storage in Ae. aegypti females is influenced by ejaculate volume, and that sperm that do not enter the spermathecae remain in the bursa, where they appear to degrade. The consistent presence of sperm in the bursa, even when males transferred low sperm quantities, suggests that the putative degradation of bursa sperm may play a role in Ae. aegypti female fertility, potentially identifying a novel process in this important vector species.