The small yellow croaker Larimichthys polyactis is an economically important marine fish in Northeast Asia.Currently,its natural resources are threatened by overfishing and environmental pollution.Therefore,research o...The small yellow croaker Larimichthys polyactis is an economically important marine fish in Northeast Asia.Currently,its natural resources are threatened by overfishing and environmental pollution.Therefore,research on the reproductive system of the fish is crucial.Here,we studied the testis development and ultrastructural features of spermatogenesis in cultured L.polyactis using anatomical,histological,and ultrastructural techniques.A pair of testes,consisting of a central sperm duct and radial seminiferous lobules,were observed.The reproduction cycle of testes can be divided into stages I–VI.March to May was confirmed as the breeding season for male L.polyactis,while April is the ideal period for artificial breeding.The male L.polyactis can attain sexual maturity within 1 year.The spermatogenesis of L.polyactis comprised spermatogonium,spermatocyte,spermatid,and mature spermatozoon.The morphology of spermatogenic cells changed obviously during spermiogenesis,including nuclear shaping,midpiece and flagellum formation.The mature sperms consist of an ellipsoidal head,a short midpiece,and a long flagellum.The anterior of the head with a kidney-shaped nucleus can be distinguished.The midpiece is located posterior to the head and includes four to six spherical mitochondria.The flagellum has irregular lateral fins.The testis of L.polyactis is an unrestricted lobular type,with cystic spermatogenesis,type II spermiogenesis,and type II spermatozoa.These features are highly similar to those of other Sciaenid species.Our findings provide useful insights into the mechanism underlying testis development and spermatogenesis of L.polyactis,which can facilitate the artificial breeding of this species.展开更多
The phosphoinositide-3-kinase/Akt(PI3K/AKT)signaling pathway is crucial for Sertoli cell development and completing spermatogenesis.Its main role is to promote proliferation and inhibit apoptosis.Many factors activate...The phosphoinositide-3-kinase/Akt(PI3K/AKT)signaling pathway is crucial for Sertoli cell development and completing spermatogenesis.Its main role is to promote proliferation and inhibit apoptosis.Many factors activate the PI3K/AKT pathway,like hormones,such as follicle stimulating hormone(FSH),androgen,estrogen,insulin to name a few.Many of these factors have receptors inside or on the surface of Sertoli cells(SCs).This review summarizes how these hormones directly regulate the PI3K/AKT signaling pathway in SCs,which in turn affects SC proliferation and differentiation.Further,hormone-mediated PI3K/AKT signaling also stimulates SC secretion,which is essential for germ cell development,suggesting an indirect role of PI3K/AKT signaling during spermatogenesis.These functions include promoting spermatogonia proliferation and differentiation,meiosis of spermatocytes,sperm maturation,and their release.This review also provides potential hints for clinically treating male infertility issues like cryptorchidism and Sertoli cell-only syndrome.展开更多
Background:Spermatogenesis is the process by which male gametes are formed from spermatogonial stem cells and it is essential for the reliable transmission of genetic information between generations.To date,the dynami...Background:Spermatogenesis is the process by which male gametes are formed from spermatogonial stem cells and it is essential for the reliable transmission of genetic information between generations.To date,the dynamic transcriptional changes of defined populations of male germ cells in pigs have not been reported.Results:To characterize the atlas of porcine spermatogenesis,we profiled the transcriptomes of~16,966 testicular cells from a 150-day-old pig testis through single-cell RNA-sequencing(scRNA-seq).The scRNA-seq analysis identified spermatogonia,spermatocytes,spermatids and three somatic cell types in porcine testes.The functional enrichment analysis demonstrated that these cell types played diverse roles in porcine spermatogenesis.The accuracy of the defined porcine germ cell types was further validated by comparing the data from scRNA-seq with those from bulk RNA-seq.Since we delineated four distinct spermatogonial subsets,we further identified CD99 and PODXL2 as novel cell surface markers for undifferentiated and differentiating spermatogonia,respectively.Conclusions:The present study has for the first time analyzed the transcriptome of male germ cells and somatic cells in porcine testes through scRNA-seq.Four subsets of spermatogonia were identified and two novel cell surface markers were discovered,which would be helpful for studies on spermatogonial differentiation in pigs.The datasets offer valuable information on porcine spermatogenesis,and pave the way for identification of key molecular markers involved in development of male germ cells.展开更多
Mammalian spermatogenesis contains three continuous and organized processes, by which spermatogonia undergo mitosis and differentiate to spermatocytes, follow on meiosis to form haploid spermatids and ultimately trans...Mammalian spermatogenesis contains three continuous and organized processes, by which spermatogonia undergo mitosis and differentiate to spermatocytes, follow on meiosis to form haploid spermatids and ultimately transform into spermatozoa. These processes require an accurately, spatially and temporally regulated gene expression patterns. The microRNAs are a novel class of post-transcriptional regulators. Cumulating evidences have demonstrated that microRNAs are expressed in a cell-specific or stage-specific manner during spermatogenesis. In this review, we focus on the roles of microRNAs in spermatogenesis. We highlight that N6-methyladenosine(m6A)is involved in the biogenesis of microRNAs and miRNA regulates the m6A modification on mRNA, and that specific mi RNAs have been exploited as potential biomarkers for the male factor infertility, which will provide insightful understanding of microRNA roles in spermatogenesis.展开更多
Background:Spermatogenesis is a cellular differentiation process that includes three major events:mitosis of spermatogonia,meiosis of spermatocytes and spermiogenesis.Steady-state spermatogenesis relies on functions o...Background:Spermatogenesis is a cellular differentiation process that includes three major events:mitosis of spermatogonia,meiosis of spermatocytes and spermiogenesis.Steady-state spermatogenesis relies on functions of spermatogonial stem cells(SSCs).Establishing and maintaining a foundational SSC pool is essential for continued spermatogenesis in mammals.Currently,our knowledge about SSC and spermatogenesis is severely limited in domestic animals.Results:In the present study,we examined transcriptomes of testes from domestic yaks at four different stages(3,5,8 and 24 months of age)and attempted to identify genes that are associated with key developmental events of spermatogenesis.Histological analyses showed that the most advanced germ cells within seminiferous tubules of testes from 3,5,8 and 24 months old yaks were gonocytes,spermatogonia,spermatocytes and elongated spermatids,respectively.RNA-sequencing(RNA-seq)analyses revealed that 11904,4381 and 2459 genes were differentially expressed during the gonocyte to spermatogonia transition,the mitosis to meiosis transition and the meiosis to post-meiosis transition.Further analyses identified a list of candidate genes than may regulate these important cellular processes.CXCR4,a previously identified SSC niche factor in mouse,was one of the up-regulated genes in the 5 months old yak testis.Results of immunohistochemical staining confirmed that CXCR4 was exclusively expressed in gonocytes and a subpopulation of spermatogonia in the yak testis.Conclusions:Together,these findings demonstrated histological changes of postnatal testis development in the domestic yak.During development of spermatogonial lineage,meiotic and haploid germ cells are supported by dynamic transcriptional regulation of gene expression.Our transcriptomic analyses provided a list of candidate genes that potentially play crucial roles in directing the establishment of SSC and spermatogenesis in yak.展开更多
Spermatogenesis is a complex differentiating developmental process in which undifferentiated spermatogonial germ cells differentiate into spermatocytes,spermatids,and finally,to mature spermatozoa.This multistage deve...Spermatogenesis is a complex differentiating developmental process in which undifferentiated spermatogonial germ cells differentiate into spermatocytes,spermatids,and finally,to mature spermatozoa.This multistage developmental process of spermatogenesis involves the expression of many male germ cell-specific long noncoding RNAs(lncRNAs)and highly regulated and specific gene expression.LncRNAs are a recently discovered large class of noncoding cellular transcripts that are still relatively unexplored.Only a few of them have postmeiotic;however,lncRNAs are involved in many cellular biological processes.The expression of lncRNAs is biologically relevant in the highly dynamic and complex program of spermatogenesis and has become a research focus in recent genome studies.This review considers the important roles and novel regulatory functions whereby lncRNAs modulate mammalian spermatogenesis.展开更多
The process of sperm production is well understood, but the studies of essential nutritional elements which are necessary for successful spermatogenesis are not deeply studied as yet. Our review focuses on integrating...The process of sperm production is well understood, but the studies of essential nutritional elements which are necessary for successful spermatogenesis are not deeply studied as yet. Our review focuses on integrating available information of various nutritional elements involved in spermatogenesis, sperm maturation and male reproductive system development, such as Zinc, Selenium, Folate, Vitamins and others. Antioxidants protect sperm from further oxidative damage during the entire sperm production. Other nutrients assist to improve sperm quality through different ways. The important roles of macronutrients like lipids, amino acids and proteins are emphasized here. These macronutrients constitute major components of the spermatozoa. Effects of nutritional elements on the development of Sertoli cells and Leydig cells, sperm motility and semen quality, capacity of capacitation and fertilization are discussed. A review of these areas will provide researchers with a better understanding of the compulsory participation of these nutrients in male reproductive processes. This review also pointed out gaps in current studies which will require further investigations.展开更多
Objective To determine the mitigating effects of sodium 4-phenylbutyrate(4-PBA) on high-fat diet(HFD)-induced spermatogenesis dysfunction. Methods Male rats(n = 30) were randomly divided into three groups: control, HF...Objective To determine the mitigating effects of sodium 4-phenylbutyrate(4-PBA) on high-fat diet(HFD)-induced spermatogenesis dysfunction. Methods Male rats(n = 30) were randomly divided into three groups: control, HFD, and 4-PBA(HFD +4-PBA). After 13 weeks, rats were euthanized. Testes and epididymis were harvested for further analysis. Sex hormones were detected, and hematoxylin and eosin staining was performed to examine the histological changes in the testes. Semen samples were collected to evaluate sperm quality. Spermatogenic cell apoptosis was detected by TUNEL assay. Results Compared with the control group, the final body weight and body weight gain were significantly higher in HFD-fed rats, while the testicle/body weight ratios were lower(P < 0.05). In HFD-fed rats, obvious pathological changes in the testicular tissue were observed. Treatment with 4-PBA attenuated HFD-induced histological damage, ameliorated the HFD-induced decrease in serum testosterone(T), and reduced the rate of testicular cell apoptosis(P < 0.05) in obese male rats. Finally, 4-PBA significantly improved semen parameters in HFD rats(P < 0.05). Conclusion HFD exposure induced detrimental effects on spermatogenesis, semen quality, serum T level, and testicular cell apoptosis in rats. Treatment with 4-PBA ameliorated HFD-induced impaired spermatogenesis via inhibition of apoptosis in rats. 4-PBA may have therapeutic value in the treatment of obesity-related impairment of spermatogenesis.展开更多
Prohibitin(PHB) participates in several biological processes including apoptosis, transcription regulation and suppression of cell proliferation in mammals. In this study, we cloned the full-length c DNA of prohibitin...Prohibitin(PHB) participates in several biological processes including apoptosis, transcription regulation and suppression of cell proliferation in mammals. In this study, we cloned the full-length c DNA of prohibitin 2(Cf-phb2) from the testis of scallop(Chlamys farreri). The deduced amino acid sequence presented a characteristic of PHB family with the PHB domain, and clustered with PHB2 of other species. Temporal and spatial expression of Cf-phb2 in testis during the reproductive cycle was detected by quantitative real-time PCR(q RT-PCR) and in situ hybridization. The expression of Cf-phb2 in the testis increased when testis developed from the resting stage to mature stage. The m RNA abundance of Cf-phb2 was the highest at mature stage, which was about 15-fold higher than that at proliferative stage. The expression of Cf-phb2 could be detected by in situ hybridization in all types of germ cells in testis, including spermatogonia, spermatocytes, spermatids and spermatozoa. The intensity of the signal increased with the spermatogenesis and was the highest in spermatids, which suggested that CF-PHB2 might affect the spermatogenesis of C. farreri.展开更多
Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during sp...Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during spermatogenesis is still un-known.In this study,the cytological features of spermatogenesis were investigated in Larimichthys crocea.In addition,the structure and function of prohibitin(PHB),which is associated with mitochondrial structure and dynamic,was also investigated.The full-length cDNA and protein(Lc-PHB)from the L.crocea phb gene(Lc-phb)contained 1625 base pairs and 271 amino acids,respec-tively.Lc-PHB had a conserved primary structure that resulted in a transmembrane,SPFH(the analogous region of proteins stomat-ins,prohibitins,flotillins and HflK/C),and coiled-coil domains.It was detected at high levels in the muscle,liver,and heart,and at intermediate levels in the testis,gill,and brain.Lc-phb mRNA expression was detected in spermatogenic cells by fluorescence in situ hybridization.An immunofluorescence assay revealed that PHB protein was localized in the mitochondria during spermatogenesis.Specifically,PHB expression was detected in the perinuclear cytoplasm of spermatogonia,spermatocytes,and spermatids in the early developmental stage,and mainly localized on one side of the nuclei in the cytoplasm of spermatids in a middle developmental stage,and finally on the sperm midpiece.Western blotting showed that PHB was located in the extracted mitochondria protein fraction but not in the cytoplasm protein fraction of testes.Conclusively,these results indicated that PHB was expressed in the mitochondria dur-ing spermatogenesis.In addition,the study explained the mitochondrial dynamic during fish spermatogenesis and proposed a possi-ble relationship among PHB,spermatogenesis,and male fertility.展开更多
Objective:To evaluate the effects of follicle-stimulating hormone (FSH) and estradiol benzoate (EB) on the recovery of spermatogenesis, histology, sexual hormones levels and testicular gene expression in testes of tam...Objective:To evaluate the effects of follicle-stimulating hormone (FSH) and estradiol benzoate (EB) on the recovery of spermatogenesis, histology, sexual hormones levels and testicular gene expression in testes of tamoxifen-injured rats.Methods:Forty adult rats were divided into eight groups in a factorial arrangement of tamoxifen and hormonal treatments. Half of the groups orally received 0.6 mg/kg tamoxifen, and 30 d later tamoxifen and no-tamoxifen groups (controls) were paired and assigned into four hormonal treatments with daily intramuscular injections for 10 consecutive days: 1 mL saline (control);7.5 IU FSH;12 μg/kg EB;and 7.5 IU FSH+12 μg/kg EB. One day after the last treatment, spermatozoa were recovered from epididymis, blood was processed for sex hormones concentration (testosterone, FSH and luteinizing hormone) and testes were processed for histology and RNA extraction for expression of genes related to apoptosis [caspase 3, inducible nitric oxide synthase (iNOS) and B-cell lymphoma-2 (Bcl-2)].Results: Control groups did not show significant changes in most parameters, but hormonal treatments decreased caspase 3 and iNOS and increased Bcl-2 expression. Tamoxifen significantly decreased counts, motility and viability of spermatozoa, Bcl-2 expression and sex hormones. It increased intertubular space, caspase 3 and iNOS expression, and induced seminiferous tubular atrophy. The hormonal treatments reverted spermatogenesis, hormonal levels and histology compared with controls, however not attaining the same sperm quality as controls.Conclusions:Tamoxifen is clearly detrimental to spermatogenesis and overall testicular structure and function, whereas hormonal therapy with FSH and EB can improve testicular function and revert tamoxifen-induced azoospermia.展开更多
Spermatogenesis is a complicated and highly regulated male gamete differentiation process that begins with the proliferation and differentiation of spermatogonia to the release of the mature spermatozoa.The autophagy-...Spermatogenesis is a complicated and highly regulated male gamete differentiation process that begins with the proliferation and differentiation of spermatogonia to the release of the mature spermatozoa.The autophagy-related gene atg7 has been reported as closely related to spermatogenesis and communication of Sertoli cell-germ cells in mice,including acrosome biogenesis,sperm flagellum development,and ectoplasmic specialization assembly.However,the function of es-ATG7 and its molecular regulatory mechanism during spermatogenesis in Crustacea remain largely unknown.Here,we cloned and identified es-atg7 from the testes of the Chinese mitten crabs Eriocheir sinensis and found that the expression of es-atg7 was relatively high in testes through semi-quantitative RT-PCR.The dynamic localization of es-ATG7 detected by immunofluorescence may convey information about its role in the spermatogenesis of E.sinensis.Furthermore,a knockdown of es-atg7 revealed that the malformed sperm with irregular sperm shape or loose nuclear cup and germ cell apoptosis were increased significantly.Accompanying this,we found an up-regulated expression of es-p53 during spermatogenesis in es-atg7 knockdown groups.Altogether,our results indicate that es-ATG7 plays an essential role during spermatogenesis of E.sinensis,and we demonstrated that es-ATG7 acts as an antagonist for p53-dependent apoptosis induction in this process.展开更多
Objective: To investigate the correlation between epidermal growth factor (EGF)/testicular epidermal growth factor receptor (EGF-R) and spermatogenesis in rat.Methods: Forty mature male Spraque-Dauley (SD) rats were r...Objective: To investigate the correlation between epidermal growth factor (EGF)/testicular epidermal growth factor receptor (EGF-R) and spermatogenesis in rat.Methods: Forty mature male Spraque-Dauley (SD) rats were randomly assigned to four groups, ten rats in each: sham operation group (SOG), sialoadenectomy group (SG), sialoadeand blood and testes were obtained on the 48th day after the operation. Serum EGF concentrations were determined by radioimmunoassay (RIA), expression of EGF-R in testes was examined by the immunohistochemical method, and the spermatogenesis was pathologically checked.Results:Serum EGF levels in SG-EGFIand SG decreased significantly when compared with those of SOG (P＜0.05 and P＜ 0.01, respectively). The testicular function of spermatogenesis showed a moderate to severe impairment in SG. The expression of EGF-R in Leydig cells decreased in SG (P＜ 0. 05). The two dosage groups of EGF replacement had different effects.There were no significant differences of EGF-R expression in testicular germ cells, Sertoli cells and Leydig cells in SOG, SG-EGFIand SG-EGFⅡ(P＞0.05).Conclusion: EGF may play an important role in the regulation of spermatogenesis. Serum EGF concentration and high expression of EGF-R in Leydig cells have a positive correlation with spermatogenic function of the testes.展开更多
The present study was conducted on some aspects of histology and morphology of gonadal development and spermatogenesis of the striped piggy fish,Pomadasys stridens from the Arabian Sea near Karachi coast.A preliminary...The present study was conducted on some aspects of histology and morphology of gonadal development and spermatogenesis of the striped piggy fish,Pomadasys stridens from the Arabian Sea near Karachi coast.A preliminary investigative study was carried on genetic-variability,morpho-histological characters to observe the differences between both gonads of both the sexes along with the size of the associated fat bodies.Both the gonads were investigated and classified in seven different stages of maturity.Testes were found asymmetrical in measurement with the right testis larger than the left one.It was found that meiotic activity and spermatid development showed the opposite relationship.The left testis showed a relatively greater activity than the right one.展开更多
Spermatogenesis is a critical part of reproduction in insects;however,its molecular mechanism is still largely unknown.In this study,we identified a testis-specific gene CG3526 in Drosophila melanogaster.Bioinformatic...Spermatogenesis is a critical part of reproduction in insects;however,its molecular mechanism is still largely unknown.In this study,we identified a testis-specific gene CG3526 in Drosophila melanogaster.Bioinformatics analysis showed that CG3526 contains a zinc binding domain and 2 C2H2 type zinc fingers,and it is clustered to the vertebrate really interesting new gene(RING)family E3 ubiquitin-protein ligases.When CG3526 was knocked down by RNA interference(RNAi),the testis became much smaller in size,and the apical tip exhibited a sharp and thin end instead of the blunt and round shape in the control testis.More importantly,compared to the control flies,only a few mature sperm were present in the seminal vesicle of C587-Gal4>CG3526 RNAi flies.Immunofluorescence staining of the testis from CG3526 RNAi flies showed that the homeostasis of testis stem cell niche was disrupted,cell distribution in the apical tip was scattered,and the process of spermatogenesis was not completed.Furthermore,we found that the phenotype of CG3526 RNAi flies’testis was similar to that of testis of Stat92E RNAi flies,the expression level of CG3526 was significantly downregulated in the Stat92EF06346 mutant flies,and the promoter activity of CG3526 was upregulated by STAT92E.Taken together,our results indicated that CG3526 is a downstream effector gene in the JAK-STAT signaling pathway that plays a key role in the spermatogenesis of Drosophila.展开更多
Male germ cells employ specific metabolic pathways throughout their developmental stages.In a previous study,we discovered heightened expression of pyruvate kinase M(PKM),a pivotal glycolytic enzyme,in spermatogonia a...Male germ cells employ specific metabolic pathways throughout their developmental stages.In a previous study,we discovered heightened expression of pyruvate kinase M(PKM),a pivotal glycolytic enzyme,in spermatogonia and spermatids.To gain deeper insights into PKM's roles in spermatogenesis,sperm function,and male fertility,we engineered a conditional-knockout mouse model(Pkm-vkO mice)to selectively disrupt the Pkm gene within germ cells.Despite maintaining regular testicular histology and sperm morphology,the male Pkm-vko mice were infertility,characterized by significant impairments in sperm motility and adenosine triphosphate(ATP)generation.In addition,Pkm-null spermatozoa exhibited similar deficits in protein tyrosine phosphorylation linked to capacitation,as well as compromised performance in in vitro fertilization experiments.To conclude,PKM's presence is not obligatory for the entirety of spermatogenesis in male germ cells;however,it emerges as a critical factor influencing sperm motility and overall male fertility.展开更多
Mammalian testis exhibits remarkably high transcriptome complexity,and spermatogenesis undergoes two periods of transcriptional cessation.These make the RNA-binding proteins(RBPs)the utmost importance during male germ...Mammalian testis exhibits remarkably high transcriptome complexity,and spermatogenesis undergoes two periods of transcriptional cessation.These make the RNA-binding proteins(RBPs)the utmost importance during male germ cell development.Heterogeneous nuclear ribonucleoproteins(hnRNPs)are a large family of RBPs implicated in many steps of RNA processing;however,their roles in spermatogenesis are largely unknown.Here,we investigated the expression pattern of 12 hnRNP family members in mouse testes and found that most detected members are highly expressed in the testis.Furthermore,we found that most of the detected hnRNP proteins(hnRNPD,hnRNPK,hnRNPQ,hnRNPU,and hnRNPUL1)display the highest signals in the nuclei of pachytene spermatocytes,round spermatids,and Sertoli cells,whereas hnRNPE1 exclusively concentrates in the manchette of elongating spermatids.The expression of these hnRNP proteins showed both similarities and specificity,suggesting their diverse roles in spermatogenesis.展开更多
Congenital bilateral absence of the vas deferens(CBAVD)is observed in 1%–2%of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator(CFTR)mutations.CFTR i...Congenital bilateral absence of the vas deferens(CBAVD)is observed in 1%–2%of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator(CFTR)mutations.CFTR is one of the most well-known genes related to male fertility.The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia(NOA).CFTR mutations are highly polymorphic and have established ethnic specificity.Compared with F508Del in Caucasians,the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis.However,whether p.G970D participates in male infertility remains unknown.Herein,a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA.Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation(4.1%,5/122),excluding polymorphic sites.Furthermore,we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients.The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells.In spermatocyte GC-2(spd)ts(GC2)Cftr p.G965D cells,RNA splicing variants were detected and CFTR expression decreased,which may contribute to the phenotypes associated with impaired spermatogenesis.Thus,this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.展开更多
Spermatogenesis is a continual process that occurs in the testes,in which diploid spermatogonial stem cells(SSCs)differentiate and generate haploid spermatozoa.This highly efficient and intricate process is orchestrat...Spermatogenesis is a continual process that occurs in the testes,in which diploid spermatogonial stem cells(SSCs)differentiate and generate haploid spermatozoa.This highly efficient and intricate process is orchestrated at multiple levels.N^(6)-methyladenosine(m^(6)A),an epigenetic modification prevalent in mRNAs,is implicated in the transcriptional regulation during spermatogenesis.However,the dynamics of m^(6)A modification in non-rodent mammalian species remains unclear.Here,we systematically investigated the profile and role of m^(6)A during spermatogenesis in pigs.By analyzing the transcriptomic distribution of m^(6)A in spermatogonia,spermatocytes,and round spermatids,we identified a globally conserved m^(6)A pattern between porcine and murine genes with spermatogenic function.We found that m^(6)A was enriched in a group of genes that specifically encode the metabolic enzymes and regulators.In addition,transcriptomes in porcine male germ cells could be subjected to the m^(6)A modification.Our data show that m^(6)A plays the regulatory roles during spermatogenesis in pigs,which is similar to that in mice.Illustrations of this point are three genes(SETDB1,FOXO1,and FOXO3)that are crucial to the determination of the fate of SSCs.To the best of our knowledge,this study for the first time uncovers the expression profile and role of m^(6)A during spermatogenesis in large animals and provides insights into the intricate transcriptional regulation underlying the lifelong male fertility in non-rodent mammalian species.展开更多
Phosphoribosyl-pyrophosphate synthetase 2(PRPS2)is a rate-limiting enzyme and plays an important role in purine and pyrimidine nucleotide synthesis.Recent studies report that PRPS2 is involved in male infertility.Howe...Phosphoribosyl-pyrophosphate synthetase 2(PRPS2)is a rate-limiting enzyme and plays an important role in purine and pyrimidine nucleotide synthesis.Recent studies report that PRPS2 is involved in male infertility.However,the role of PRPS2 in hypospermatogenesis is unknown.In this study,the relationship of PRPS2 with hypospermatogenesis and spermatogenic cell apoptosis was investigated.The results showed that PRPS2 depletion increased the number of apoptotic spermatogenic cells in vitro.PRPS2 was downregulated in a mouse model of hypospermatogenesis.When PRPS2 expression was knocked down in mouse testes,hypospermatogenesis and accelerated apoptosis of spermatogenic cells were noted.E2F transcription factor 1(E2F1)was confirmed as the target gene of PRPS2 and played a key role in cell apoptosis by regulating the P53/Bcl-xl/Bcl-2/Caspase 6/Caspase 9 apoptosis pathway.Therefore,these data indicate that PRPS2 depletion contributes to the apoptosis of spermatogenic cells and is associated with hypospermatogenesis,which may be helpful for the diagnosis of male infertility.展开更多
基金the NSFC-Zhejiang Joint Fund for the Integration of Industrialization and Informatization(No.U1809212)the Scientific and Technical Project of Zhejiang Province(Nos.2021C02055,2017C02013)+2 种基金the National Natural Science Foundation of China(No.31272642)Healthy Aquaculture,the K.C.Wong Magna Fund in Ningbo Universitythe Collaborative Innovation Center for Zhejiang Marine High-Efficiency。
文摘The small yellow croaker Larimichthys polyactis is an economically important marine fish in Northeast Asia.Currently,its natural resources are threatened by overfishing and environmental pollution.Therefore,research on the reproductive system of the fish is crucial.Here,we studied the testis development and ultrastructural features of spermatogenesis in cultured L.polyactis using anatomical,histological,and ultrastructural techniques.A pair of testes,consisting of a central sperm duct and radial seminiferous lobules,were observed.The reproduction cycle of testes can be divided into stages I–VI.March to May was confirmed as the breeding season for male L.polyactis,while April is the ideal period for artificial breeding.The male L.polyactis can attain sexual maturity within 1 year.The spermatogenesis of L.polyactis comprised spermatogonium,spermatocyte,spermatid,and mature spermatozoon.The morphology of spermatogenic cells changed obviously during spermiogenesis,including nuclear shaping,midpiece and flagellum formation.The mature sperms consist of an ellipsoidal head,a short midpiece,and a long flagellum.The anterior of the head with a kidney-shaped nucleus can be distinguished.The midpiece is located posterior to the head and includes four to six spherical mitochondria.The flagellum has irregular lateral fins.The testis of L.polyactis is an unrestricted lobular type,with cystic spermatogenesis,type II spermiogenesis,and type II spermatozoa.These features are highly similar to those of other Sciaenid species.Our findings provide useful insights into the mechanism underlying testis development and spermatogenesis of L.polyactis,which can facilitate the artificial breeding of this species.
基金supported in part by the National Natural Science Foundation of China(Nos.32270555 and 32072954).
文摘The phosphoinositide-3-kinase/Akt(PI3K/AKT)signaling pathway is crucial for Sertoli cell development and completing spermatogenesis.Its main role is to promote proliferation and inhibit apoptosis.Many factors activate the PI3K/AKT pathway,like hormones,such as follicle stimulating hormone(FSH),androgen,estrogen,insulin to name a few.Many of these factors have receptors inside or on the surface of Sertoli cells(SCs).This review summarizes how these hormones directly regulate the PI3K/AKT signaling pathway in SCs,which in turn affects SC proliferation and differentiation.Further,hormone-mediated PI3K/AKT signaling also stimulates SC secretion,which is essential for germ cell development,suggesting an indirect role of PI3K/AKT signaling during spermatogenesis.These functions include promoting spermatogonia proliferation and differentiation,meiosis of spermatocytes,sperm maturation,and their release.This review also provides potential hints for clinically treating male infertility issues like cryptorchidism and Sertoli cell-only syndrome.
基金This study was supported in part by the National Natural Science Foundation of China(Grant No.31772605)to WXZResearch Project of Shaanxi Science and Technology Department(2020NY-003)to TZ.
文摘Background:Spermatogenesis is the process by which male gametes are formed from spermatogonial stem cells and it is essential for the reliable transmission of genetic information between generations.To date,the dynamic transcriptional changes of defined populations of male germ cells in pigs have not been reported.Results:To characterize the atlas of porcine spermatogenesis,we profiled the transcriptomes of~16,966 testicular cells from a 150-day-old pig testis through single-cell RNA-sequencing(scRNA-seq).The scRNA-seq analysis identified spermatogonia,spermatocytes,spermatids and three somatic cell types in porcine testes.The functional enrichment analysis demonstrated that these cell types played diverse roles in porcine spermatogenesis.The accuracy of the defined porcine germ cell types was further validated by comparing the data from scRNA-seq with those from bulk RNA-seq.Since we delineated four distinct spermatogonial subsets,we further identified CD99 and PODXL2 as novel cell surface markers for undifferentiated and differentiating spermatogonia,respectively.Conclusions:The present study has for the first time analyzed the transcriptome of male germ cells and somatic cells in porcine testes through scRNA-seq.Four subsets of spermatogonia were identified and two novel cell surface markers were discovered,which would be helpful for studies on spermatogonial differentiation in pigs.The datasets offer valuable information on porcine spermatogenesis,and pave the way for identification of key molecular markers involved in development of male germ cells.
基金funded,in part,by the National Basic Research Program of China(973 programNo.2013CB943103)+1 种基金the National Natural Science Foundation of China(No.31272439,No.31230048 and No.31572401)Programs Foundation of Ministry of Education of China(No.20130204110017)
文摘Mammalian spermatogenesis contains three continuous and organized processes, by which spermatogonia undergo mitosis and differentiate to spermatocytes, follow on meiosis to form haploid spermatids and ultimately transform into spermatozoa. These processes require an accurately, spatially and temporally regulated gene expression patterns. The microRNAs are a novel class of post-transcriptional regulators. Cumulating evidences have demonstrated that microRNAs are expressed in a cell-specific or stage-specific manner during spermatogenesis. In this review, we focus on the roles of microRNAs in spermatogenesis. We highlight that N6-methyladenosine(m6A)is involved in the biogenesis of microRNAs and miRNA regulates the m6A modification on mRNA, and that specific mi RNAs have been exploited as potential biomarkers for the male factor infertility, which will provide insightful understanding of microRNA roles in spermatogenesis.
基金supported by National Key Research&Development Project grant(2016YFC0501805)Qinghai Department of Science and Technology grants(2017-NK-154 and 2016-ZJ-917Q)+2 种基金a STS grant from Chinese Academy of Sciences(KFJ-STS-QYZD-113)supported by the CAS“100 Talents” and Qinghai “1000 Talents” programsfunded by CAS “Light of West China Foundation”
文摘Background:Spermatogenesis is a cellular differentiation process that includes three major events:mitosis of spermatogonia,meiosis of spermatocytes and spermiogenesis.Steady-state spermatogenesis relies on functions of spermatogonial stem cells(SSCs).Establishing and maintaining a foundational SSC pool is essential for continued spermatogenesis in mammals.Currently,our knowledge about SSC and spermatogenesis is severely limited in domestic animals.Results:In the present study,we examined transcriptomes of testes from domestic yaks at four different stages(3,5,8 and 24 months of age)and attempted to identify genes that are associated with key developmental events of spermatogenesis.Histological analyses showed that the most advanced germ cells within seminiferous tubules of testes from 3,5,8 and 24 months old yaks were gonocytes,spermatogonia,spermatocytes and elongated spermatids,respectively.RNA-sequencing(RNA-seq)analyses revealed that 11904,4381 and 2459 genes were differentially expressed during the gonocyte to spermatogonia transition,the mitosis to meiosis transition and the meiosis to post-meiosis transition.Further analyses identified a list of candidate genes than may regulate these important cellular processes.CXCR4,a previously identified SSC niche factor in mouse,was one of the up-regulated genes in the 5 months old yak testis.Results of immunohistochemical staining confirmed that CXCR4 was exclusively expressed in gonocytes and a subpopulation of spermatogonia in the yak testis.Conclusions:Together,these findings demonstrated histological changes of postnatal testis development in the domestic yak.During development of spermatogonial lineage,meiotic and haploid germ cells are supported by dynamic transcriptional regulation of gene expression.Our transcriptomic analyses provided a list of candidate genes that potentially play crucial roles in directing the establishment of SSC and spermatogenesis in yak.
基金supported by the National Natural Science Foundation of China(3187131067)the Agricultural Science and Technology Innovation Program(ASTIP-2016-IAS-06 and CAAS-XTCX2016010)the China Agriculture Research System(CARS-36).
文摘Spermatogenesis is a complex differentiating developmental process in which undifferentiated spermatogonial germ cells differentiate into spermatocytes,spermatids,and finally,to mature spermatozoa.This multistage developmental process of spermatogenesis involves the expression of many male germ cell-specific long noncoding RNAs(lncRNAs)and highly regulated and specific gene expression.LncRNAs are a recently discovered large class of noncoding cellular transcripts that are still relatively unexplored.Only a few of them have postmeiotic;however,lncRNAs are involved in many cellular biological processes.The expression of lncRNAs is biologically relevant in the highly dynamic and complex program of spermatogenesis and has become a research focus in recent genome studies.This review considers the important roles and novel regulatory functions whereby lncRNAs modulate mammalian spermatogenesis.
文摘The process of sperm production is well understood, but the studies of essential nutritional elements which are necessary for successful spermatogenesis are not deeply studied as yet. Our review focuses on integrating available information of various nutritional elements involved in spermatogenesis, sperm maturation and male reproductive system development, such as Zinc, Selenium, Folate, Vitamins and others. Antioxidants protect sperm from further oxidative damage during the entire sperm production. Other nutrients assist to improve sperm quality through different ways. The important roles of macronutrients like lipids, amino acids and proteins are emphasized here. These macronutrients constitute major components of the spermatozoa. Effects of nutritional elements on the development of Sertoli cells and Leydig cells, sperm motility and semen quality, capacity of capacitation and fertilization are discussed. A review of these areas will provide researchers with a better understanding of the compulsory participation of these nutrients in male reproductive processes. This review also pointed out gaps in current studies which will require further investigations.
基金supported by the National Natural Science Foundation of China [Grant No.81703230]the Key Scientific Research Project of Universities in Henan Province [Grant No.16B330001 and No.17A330005]
文摘Objective To determine the mitigating effects of sodium 4-phenylbutyrate(4-PBA) on high-fat diet(HFD)-induced spermatogenesis dysfunction. Methods Male rats(n = 30) were randomly divided into three groups: control, HFD, and 4-PBA(HFD +4-PBA). After 13 weeks, rats were euthanized. Testes and epididymis were harvested for further analysis. Sex hormones were detected, and hematoxylin and eosin staining was performed to examine the histological changes in the testes. Semen samples were collected to evaluate sperm quality. Spermatogenic cell apoptosis was detected by TUNEL assay. Results Compared with the control group, the final body weight and body weight gain were significantly higher in HFD-fed rats, while the testicle/body weight ratios were lower(P < 0.05). In HFD-fed rats, obvious pathological changes in the testicular tissue were observed. Treatment with 4-PBA attenuated HFD-induced histological damage, ameliorated the HFD-induced decrease in serum testosterone(T), and reduced the rate of testicular cell apoptosis(P < 0.05) in obese male rats. Finally, 4-PBA significantly improved semen parameters in HFD rats(P < 0.05). Conclusion HFD exposure induced detrimental effects on spermatogenesis, semen quality, serum T level, and testicular cell apoptosis in rats. Treatment with 4-PBA ameliorated HFD-induced impaired spermatogenesis via inhibition of apoptosis in rats. 4-PBA may have therapeutic value in the treatment of obesity-related impairment of spermatogenesis.
基金supported by the National High Technology Research and Development Program of China(863 Program)(2012AA10A402)the Natural Science Foundation of Qingdao(11-2-4-1(10)-jch)
文摘Prohibitin(PHB) participates in several biological processes including apoptosis, transcription regulation and suppression of cell proliferation in mammals. In this study, we cloned the full-length c DNA of prohibitin 2(Cf-phb2) from the testis of scallop(Chlamys farreri). The deduced amino acid sequence presented a characteristic of PHB family with the PHB domain, and clustered with PHB2 of other species. Temporal and spatial expression of Cf-phb2 in testis during the reproductive cycle was detected by quantitative real-time PCR(q RT-PCR) and in situ hybridization. The expression of Cf-phb2 in the testis increased when testis developed from the resting stage to mature stage. The m RNA abundance of Cf-phb2 was the highest at mature stage, which was about 15-fold higher than that at proliferative stage. The expression of Cf-phb2 could be detected by in situ hybridization in all types of germ cells in testis, including spermatogonia, spermatocytes, spermatids and spermatozoa. The intensity of the signal increased with the spermatogenesis and was the highest in spermatids, which suggested that CF-PHB2 might affect the spermatogenesis of C. farreri.
基金funded by the Natural Science Foun dation of Zhejiang Province(No.LY18C190007)the Na tural Science Foundation of China-Zhejiang Joint Fund for the Integration of Industrialization and Informatization(No.U1809212)+3 种基金the Natural Science Foundation of Ningbo City(No.2018A610228)the Scientific and Technical Project of Zhejiang Province(No.2021C02069-1)the Scientific and Technical Project of Ningbo City(No.2021Z002),the Scientific Research Foundation of Ningbo University(No.XYL19023)the Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture,the K.C.Wong Magna Fund in Ningbo University.
文摘Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during spermatogenesis is still un-known.In this study,the cytological features of spermatogenesis were investigated in Larimichthys crocea.In addition,the structure and function of prohibitin(PHB),which is associated with mitochondrial structure and dynamic,was also investigated.The full-length cDNA and protein(Lc-PHB)from the L.crocea phb gene(Lc-phb)contained 1625 base pairs and 271 amino acids,respec-tively.Lc-PHB had a conserved primary structure that resulted in a transmembrane,SPFH(the analogous region of proteins stomat-ins,prohibitins,flotillins and HflK/C),and coiled-coil domains.It was detected at high levels in the muscle,liver,and heart,and at intermediate levels in the testis,gill,and brain.Lc-phb mRNA expression was detected in spermatogenic cells by fluorescence in situ hybridization.An immunofluorescence assay revealed that PHB protein was localized in the mitochondria during spermatogenesis.Specifically,PHB expression was detected in the perinuclear cytoplasm of spermatogonia,spermatocytes,and spermatids in the early developmental stage,and mainly localized on one side of the nuclei in the cytoplasm of spermatids in a middle developmental stage,and finally on the sperm midpiece.Western blotting showed that PHB was located in the extracted mitochondria protein fraction but not in the cytoplasm protein fraction of testes.Conclusively,these results indicated that PHB was expressed in the mitochondria dur-ing spermatogenesis.In addition,the study explained the mitochondrial dynamic during fish spermatogenesis and proposed a possi-ble relationship among PHB,spermatogenesis,and male fertility.
文摘Objective:To evaluate the effects of follicle-stimulating hormone (FSH) and estradiol benzoate (EB) on the recovery of spermatogenesis, histology, sexual hormones levels and testicular gene expression in testes of tamoxifen-injured rats.Methods:Forty adult rats were divided into eight groups in a factorial arrangement of tamoxifen and hormonal treatments. Half of the groups orally received 0.6 mg/kg tamoxifen, and 30 d later tamoxifen and no-tamoxifen groups (controls) were paired and assigned into four hormonal treatments with daily intramuscular injections for 10 consecutive days: 1 mL saline (control);7.5 IU FSH;12 μg/kg EB;and 7.5 IU FSH+12 μg/kg EB. One day after the last treatment, spermatozoa were recovered from epididymis, blood was processed for sex hormones concentration (testosterone, FSH and luteinizing hormone) and testes were processed for histology and RNA extraction for expression of genes related to apoptosis [caspase 3, inducible nitric oxide synthase (iNOS) and B-cell lymphoma-2 (Bcl-2)].Results: Control groups did not show significant changes in most parameters, but hormonal treatments decreased caspase 3 and iNOS and increased Bcl-2 expression. Tamoxifen significantly decreased counts, motility and viability of spermatozoa, Bcl-2 expression and sex hormones. It increased intertubular space, caspase 3 and iNOS expression, and induced seminiferous tubular atrophy. The hormonal treatments reverted spermatogenesis, hormonal levels and histology compared with controls, however not attaining the same sperm quality as controls.Conclusions:Tamoxifen is clearly detrimental to spermatogenesis and overall testicular structure and function, whereas hormonal therapy with FSH and EB can improve testicular function and revert tamoxifen-induced azoospermia.
基金supported by the National Natural Science Foundation of China(No.32072954 and No.41776144)Zhejiang Province Public Welfare Technology Application Research Project(including Natural Science Foundation)(No.LGF20C120001).
文摘Spermatogenesis is a complicated and highly regulated male gamete differentiation process that begins with the proliferation and differentiation of spermatogonia to the release of the mature spermatozoa.The autophagy-related gene atg7 has been reported as closely related to spermatogenesis and communication of Sertoli cell-germ cells in mice,including acrosome biogenesis,sperm flagellum development,and ectoplasmic specialization assembly.However,the function of es-ATG7 and its molecular regulatory mechanism during spermatogenesis in Crustacea remain largely unknown.Here,we cloned and identified es-atg7 from the testes of the Chinese mitten crabs Eriocheir sinensis and found that the expression of es-atg7 was relatively high in testes through semi-quantitative RT-PCR.The dynamic localization of es-ATG7 detected by immunofluorescence may convey information about its role in the spermatogenesis of E.sinensis.Furthermore,a knockdown of es-atg7 revealed that the malformed sperm with irregular sperm shape or loose nuclear cup and germ cell apoptosis were increased significantly.Accompanying this,we found an up-regulated expression of es-p53 during spermatogenesis in es-atg7 knockdown groups.Altogether,our results indicate that es-ATG7 plays an essential role during spermatogenesis of E.sinensis,and we demonstrated that es-ATG7 acts as an antagonist for p53-dependent apoptosis induction in this process.
基金The project supported by Anhui Provincial Natural Science Foundation (99044554), China.
文摘Objective: To investigate the correlation between epidermal growth factor (EGF)/testicular epidermal growth factor receptor (EGF-R) and spermatogenesis in rat.Methods: Forty mature male Spraque-Dauley (SD) rats were randomly assigned to four groups, ten rats in each: sham operation group (SOG), sialoadenectomy group (SG), sialoadeand blood and testes were obtained on the 48th day after the operation. Serum EGF concentrations were determined by radioimmunoassay (RIA), expression of EGF-R in testes was examined by the immunohistochemical method, and the spermatogenesis was pathologically checked.Results:Serum EGF levels in SG-EGFIand SG decreased significantly when compared with those of SOG (P＜0.05 and P＜ 0.01, respectively). The testicular function of spermatogenesis showed a moderate to severe impairment in SG. The expression of EGF-R in Leydig cells decreased in SG (P＜ 0. 05). The two dosage groups of EGF replacement had different effects.There were no significant differences of EGF-R expression in testicular germ cells, Sertoli cells and Leydig cells in SOG, SG-EGFIand SG-EGFⅡ(P＞0.05).Conclusion: EGF may play an important role in the regulation of spermatogenesis. Serum EGF concentration and high expression of EGF-R in Leydig cells have a positive correlation with spermatogenic function of the testes.
文摘The present study was conducted on some aspects of histology and morphology of gonadal development and spermatogenesis of the striped piggy fish,Pomadasys stridens from the Arabian Sea near Karachi coast.A preliminary investigative study was carried on genetic-variability,morpho-histological characters to observe the differences between both gonads of both the sexes along with the size of the associated fat bodies.Both the gonads were investigated and classified in seven different stages of maturity.Testes were found asymmetrical in measurement with the right testis larger than the left one.It was found that meiotic activity and spermatid development showed the opposite relationship.The left testis showed a relatively greater activity than the right one.
基金This work was supported by the National Natural Science Foundation of China(No.31970474 and No.32272606)China Postdoctoral Science Foundation(No.2022M721219).
文摘Spermatogenesis is a critical part of reproduction in insects;however,its molecular mechanism is still largely unknown.In this study,we identified a testis-specific gene CG3526 in Drosophila melanogaster.Bioinformatics analysis showed that CG3526 contains a zinc binding domain and 2 C2H2 type zinc fingers,and it is clustered to the vertebrate really interesting new gene(RING)family E3 ubiquitin-protein ligases.When CG3526 was knocked down by RNA interference(RNAi),the testis became much smaller in size,and the apical tip exhibited a sharp and thin end instead of the blunt and round shape in the control testis.More importantly,compared to the control flies,only a few mature sperm were present in the seminal vesicle of C587-Gal4>CG3526 RNAi flies.Immunofluorescence staining of the testis from CG3526 RNAi flies showed that the homeostasis of testis stem cell niche was disrupted,cell distribution in the apical tip was scattered,and the process of spermatogenesis was not completed.Furthermore,we found that the phenotype of CG3526 RNAi flies’testis was similar to that of testis of Stat92E RNAi flies,the expression level of CG3526 was significantly downregulated in the Stat92EF06346 mutant flies,and the promoter activity of CG3526 was upregulated by STAT92E.Taken together,our results indicated that CG3526 is a downstream effector gene in the JAK-STAT signaling pathway that plays a key role in the spermatogenesis of Drosophila.
基金supported by the National Natural Science Foundation of China(No.81571488 and No.81771637).
文摘Male germ cells employ specific metabolic pathways throughout their developmental stages.In a previous study,we discovered heightened expression of pyruvate kinase M(PKM),a pivotal glycolytic enzyme,in spermatogonia and spermatids.To gain deeper insights into PKM's roles in spermatogenesis,sperm function,and male fertility,we engineered a conditional-knockout mouse model(Pkm-vkO mice)to selectively disrupt the Pkm gene within germ cells.Despite maintaining regular testicular histology and sperm morphology,the male Pkm-vko mice were infertility,characterized by significant impairments in sperm motility and adenosine triphosphate(ATP)generation.In addition,Pkm-null spermatozoa exhibited similar deficits in protein tyrosine phosphorylation linked to capacitation,as well as compromised performance in in vitro fertilization experiments.To conclude,PKM's presence is not obligatory for the entirety of spermatogenesis in male germ cells;however,it emerges as a critical factor influencing sperm motility and overall male fertility.
基金This work,in part,was supported by grants from the National Natural Science Foundation of China(31801237 to XLW and 82171605 to SQY)the Science Technology and Innovation Commission of Shenzhen Municipality(JCYJ20170818160910316 to SQY).
文摘Mammalian testis exhibits remarkably high transcriptome complexity,and spermatogenesis undergoes two periods of transcriptional cessation.These make the RNA-binding proteins(RBPs)the utmost importance during male germ cell development.Heterogeneous nuclear ribonucleoproteins(hnRNPs)are a large family of RBPs implicated in many steps of RNA processing;however,their roles in spermatogenesis are largely unknown.Here,we investigated the expression pattern of 12 hnRNP family members in mouse testes and found that most detected members are highly expressed in the testis.Furthermore,we found that most of the detected hnRNP proteins(hnRNPD,hnRNPK,hnRNPQ,hnRNPU,and hnRNPUL1)display the highest signals in the nuclei of pachytene spermatocytes,round spermatids,and Sertoli cells,whereas hnRNPE1 exclusively concentrates in the manchette of elongating spermatids.The expression of these hnRNP proteins showed both similarities and specificity,suggesting their diverse roles in spermatogenesis.
基金support from the National Key Research and Developmental Program of China (No.2018YFC1003603)the National Natural Science Foundation of China (No.81971445).
文摘Congenital bilateral absence of the vas deferens(CBAVD)is observed in 1%–2%of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator(CFTR)mutations.CFTR is one of the most well-known genes related to male fertility.The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia(NOA).CFTR mutations are highly polymorphic and have established ethnic specificity.Compared with F508Del in Caucasians,the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis.However,whether p.G970D participates in male infertility remains unknown.Herein,a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA.Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation(4.1%,5/122),excluding polymorphic sites.Furthermore,we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients.The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells.In spermatocyte GC-2(spd)ts(GC2)Cftr p.G965D cells,RNA splicing variants were detected and CFTR expression decreased,which may contribute to the phenotypes associated with impaired spermatogenesis.Thus,this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.
基金the National Natural Science Foundation of China(Grant No.31572401)to Wenxian Zengthe Research Project of Shaanxi Science and Technology Department(Grant No.2020NY-003)to Tao Zhangthe National Natural Science Foundation of China(Grant No.81703193)to Yinghua Lv.
文摘Spermatogenesis is a continual process that occurs in the testes,in which diploid spermatogonial stem cells(SSCs)differentiate and generate haploid spermatozoa.This highly efficient and intricate process is orchestrated at multiple levels.N^(6)-methyladenosine(m^(6)A),an epigenetic modification prevalent in mRNAs,is implicated in the transcriptional regulation during spermatogenesis.However,the dynamics of m^(6)A modification in non-rodent mammalian species remains unclear.Here,we systematically investigated the profile and role of m^(6)A during spermatogenesis in pigs.By analyzing the transcriptomic distribution of m^(6)A in spermatogonia,spermatocytes,and round spermatids,we identified a globally conserved m^(6)A pattern between porcine and murine genes with spermatogenic function.We found that m^(6)A was enriched in a group of genes that specifically encode the metabolic enzymes and regulators.In addition,transcriptomes in porcine male germ cells could be subjected to the m^(6)A modification.Our data show that m^(6)A plays the regulatory roles during spermatogenesis in pigs,which is similar to that in mice.Illustrations of this point are three genes(SETDB1,FOXO1,and FOXO3)that are crucial to the determination of the fate of SSCs.To the best of our knowledge,this study for the first time uncovers the expression profile and role of m^(6)A during spermatogenesis in large animals and provides insights into the intricate transcriptional regulation underlying the lifelong male fertility in non-rodent mammalian species.
基金the Science and Technology Projects of Shenzhen(No.JCYJ20160428173152329)the Fundamental Research Funds for the Central Universities(No.21617316)+1 种基金the Guangdong Medical Research Fund(No.A2019553)the National Natural Science Foundation of China(No.81773277).
文摘Phosphoribosyl-pyrophosphate synthetase 2(PRPS2)is a rate-limiting enzyme and plays an important role in purine and pyrimidine nucleotide synthesis.Recent studies report that PRPS2 is involved in male infertility.However,the role of PRPS2 in hypospermatogenesis is unknown.In this study,the relationship of PRPS2 with hypospermatogenesis and spermatogenic cell apoptosis was investigated.The results showed that PRPS2 depletion increased the number of apoptotic spermatogenic cells in vitro.PRPS2 was downregulated in a mouse model of hypospermatogenesis.When PRPS2 expression was knocked down in mouse testes,hypospermatogenesis and accelerated apoptosis of spermatogenic cells were noted.E2F transcription factor 1(E2F1)was confirmed as the target gene of PRPS2 and played a key role in cell apoptosis by regulating the P53/Bcl-xl/Bcl-2/Caspase 6/Caspase 9 apoptosis pathway.Therefore,these data indicate that PRPS2 depletion contributes to the apoptosis of spermatogenic cells and is associated with hypospermatogenesis,which may be helpful for the diagnosis of male infertility.
