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
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
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
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 complex process regulated by endocrine and testicular paracrine/autocrine factors.Gonadotropins are involved in the regulation of several testicular paracrine factors, mainly of the IL-1 family an...Spermatogenesis is a complex process regulated by endocrine and testicular paracrine/autocrine factors.Gonadotropins are involved in the regulation of several testicular paracrine factors, mainly of the IL-1 family and testicular hormones. Testicular cytokines and growth factors (such as IL-1, IL-6, TNF, IFN-T, LIF and SCF) were shown to affect both the germ cell proliferation and the Leydig and Sertoli cells functions and secretion. Cytokines and growth factors are produced by immune cells and in the interstitial and seminiferous tubular compartments by various testicular cells, including Sertoli, Leydig, peritubular cells, spermatogonia, differentiated spermatogonia and even spermatozoa. Corresponding cytokine and growth factor receptors were demonstrated on some of the testicular cells. These cytokines also control the secretion of the gonadotropins and testosterone in the testis. Under pathological conditions the levels of pro-inflammatory cytokines are increased and negatively affected spermatogenesis. Thus,the expression levels and the mechanisms involved in the regulation of testicular paracrine/autocrine factors should be considered in future therapeutic strategies for male infertility. (Asian J Androl 2004 Sep; 6: 259-268)展开更多
Aim: To determine the effect of oral administration of an aqueous extract from the roots of Lepidium meyenii (maca)on spermatogenesis in adult male rats. Methods: Male rats received an aqueous extract of the root (66....Aim: To determine the effect of oral administration of an aqueous extract from the roots of Lepidium meyenii (maca)on spermatogenesis in adult male rats. Methods: Male rats received an aqueous extract of the root (66.7 mg in one mL) twice a day for 14 consecutive days. Results: Treatment with Lepidium meyenii resulted in an increase in the weights of testis and epididymis but not the seminal vesicle weight. The length and frequency of stages IX-XIV seminiferous tubules, where mitosis occurred, were increased and stages Ⅰ-Ⅵ were reduced in rats treated with Lepidium meyenii. Conclusion: The Lepidium meyenii root invigorates spermatogenesis in male rats by acting on its initial stages (IX-XIV).展开更多
Aim:To investigate the effect of arsenic on spermatogenesis.Methods:Mature(4 months old)Wistar rats were intraperitoneally administered sodium arsenite at doses of 4,5 or 6mg·kg^-1·day^-1 for 26 days.Differe...Aim:To investigate the effect of arsenic on spermatogenesis.Methods:Mature(4 months old)Wistar rats were intraperitoneally administered sodium arsenite at doses of 4,5 or 6mg·kg^-1·day^-1 for 26 days.Different varieties of germ cells at stage Ⅶ seminiferous epithelium cycle,namely,type A spermatogonia(ASg),preleptotene spermatocytes(pLSc),midpachytene spermatocytes(mPSc) and step 7 spermatids(7Sd) were quantitatively evaluated, along with radioimmunoassay of plasma follicle-stimulating hormone(FSH),lutuneizing hormone(LH),testosterone and assessment of the epididymal sperm count.Results:In the 5 and 6 mg/kg groups,there were significant dosedependent decreases in the accessory sex organ weights,epididymal sperm count and plasma concentrations of LH,FSH and testosterone with massive degeneration of all the germ cells at stage Ⅶ,The changes were insignificant in the 4 mg/kg group.Conclusion:Arsenite has a suppressive influence on spermatogenesis and gonadotrophin and testosterone release in rats.展开更多
Aim: To identify and characterize a novel gene with potential roles in testis development and spermatogenesis.Methods: A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized wit...Aim: To identify and characterize a novel gene with potential roles in testis development and spermatogenesis.Methods: A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes. Differentially expressed genes were isolated and sequenced. RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis. A range of bioinformatical programs including Gene Runner, SMART, NCBI Blast and Emboss CpGPlot were used to characterize the new gene's feature. Results: A novel testis-specific gene,NYD-SPS, was differentially expressed in fetal and adult testes. The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved lie and Gin residues), a Carbamate kinase-like domain, a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain. RT-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined. In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes. Conclusion: NYD-SP5 is a newly found testisspecific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.展开更多
Aim: To study the effect of testosterone undecanoate (TU) injection on spermatogenesis in rats. Methods: Twenty adult SD rats received vehicle or TU (8 mg/kg, 19 mg/kg or 025 mg/kg) injection, im, every 15 days for da...Aim: To study the effect of testosterone undecanoate (TU) injection on spermatogenesis in rats. Methods: Twenty adult SD rats received vehicle or TU (8 mg/kg, 19 mg/kg or 025 mg/kg) injection, im, every 15 days for days, and another 38 animals received similar treatments for 130 days with half of them undergoing a recovery phase of 120 days (5 rats for each treatment). At the end of the treatment, testes were removed and the diameter of the seminiferous tubules and the number of late elongated spermatids ( steps 15 - 19 ) per testis were estimated with stereological methods as a measure of the spermatogenic efficiency. Results: Low dose (8 mg/kg) TU treatment virtually had no effect on spermatogenesis. A dose of 19 mg/kg slightly suppressed spermatogenesis 60 days after treatment, and severe suppression occurred after another 70 days of dosing. Spermatogenesis was completely recovered at the end of the recovery phase. Large dose (625 mg/kg) TU treatment did not significantly affect spermatogenesis and was well tolerated by animals. Conclusion: TU injection reversibly suppresses spermatogenesis in rats.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.
文摘Spermatogenesis is a complex process regulated by endocrine and testicular paracrine/autocrine factors.Gonadotropins are involved in the regulation of several testicular paracrine factors, mainly of the IL-1 family and testicular hormones. Testicular cytokines and growth factors (such as IL-1, IL-6, TNF, IFN-T, LIF and SCF) were shown to affect both the germ cell proliferation and the Leydig and Sertoli cells functions and secretion. Cytokines and growth factors are produced by immune cells and in the interstitial and seminiferous tubular compartments by various testicular cells, including Sertoli, Leydig, peritubular cells, spermatogonia, differentiated spermatogonia and even spermatozoa. Corresponding cytokine and growth factor receptors were demonstrated on some of the testicular cells. These cytokines also control the secretion of the gonadotropins and testosterone in the testis. Under pathological conditions the levels of pro-inflammatory cytokines are increased and negatively affected spermatogenesis. Thus,the expression levels and the mechanisms involved in the regulation of testicular paracrine/autocrine factors should be considered in future therapeutic strategies for male infertility. (Asian J Androl 2004 Sep; 6: 259-268)
文摘Aim: To determine the effect of oral administration of an aqueous extract from the roots of Lepidium meyenii (maca)on spermatogenesis in adult male rats. Methods: Male rats received an aqueous extract of the root (66.7 mg in one mL) twice a day for 14 consecutive days. Results: Treatment with Lepidium meyenii resulted in an increase in the weights of testis and epididymis but not the seminal vesicle weight. The length and frequency of stages IX-XIV seminiferous tubules, where mitosis occurred, were increased and stages Ⅰ-Ⅵ were reduced in rats treated with Lepidium meyenii. Conclusion: The Lepidium meyenii root invigorates spermatogenesis in male rats by acting on its initial stages (IX-XIV).
文摘Aim:To investigate the effect of arsenic on spermatogenesis.Methods:Mature(4 months old)Wistar rats were intraperitoneally administered sodium arsenite at doses of 4,5 or 6mg·kg^-1·day^-1 for 26 days.Different varieties of germ cells at stage Ⅶ seminiferous epithelium cycle,namely,type A spermatogonia(ASg),preleptotene spermatocytes(pLSc),midpachytene spermatocytes(mPSc) and step 7 spermatids(7Sd) were quantitatively evaluated, along with radioimmunoassay of plasma follicle-stimulating hormone(FSH),lutuneizing hormone(LH),testosterone and assessment of the epididymal sperm count.Results:In the 5 and 6 mg/kg groups,there were significant dosedependent decreases in the accessory sex organ weights,epididymal sperm count and plasma concentrations of LH,FSH and testosterone with massive degeneration of all the germ cells at stage Ⅶ,The changes were insignificant in the 4 mg/kg group.Conclusion:Arsenite has a suppressive influence on spermatogenesis and gonadotrophin and testosterone release in rats.
文摘Aim: To identify and characterize a novel gene with potential roles in testis development and spermatogenesis.Methods: A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes. Differentially expressed genes were isolated and sequenced. RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis. A range of bioinformatical programs including Gene Runner, SMART, NCBI Blast and Emboss CpGPlot were used to characterize the new gene's feature. Results: A novel testis-specific gene,NYD-SPS, was differentially expressed in fetal and adult testes. The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved lie and Gin residues), a Carbamate kinase-like domain, a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain. RT-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined. In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes. Conclusion: NYD-SP5 is a newly found testisspecific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.
基金Financially supported by a"9th five-year" National Key Grant of Science and Technology (Grant number:969040401),and by Sichuan Committee of Education.
文摘Aim: To study the effect of testosterone undecanoate (TU) injection on spermatogenesis in rats. Methods: Twenty adult SD rats received vehicle or TU (8 mg/kg, 19 mg/kg or 025 mg/kg) injection, im, every 15 days for days, and another 38 animals received similar treatments for 130 days with half of them undergoing a recovery phase of 120 days (5 rats for each treatment). At the end of the treatment, testes were removed and the diameter of the seminiferous tubules and the number of late elongated spermatids ( steps 15 - 19 ) per testis were estimated with stereological methods as a measure of the spermatogenic efficiency. Results: Low dose (8 mg/kg) TU treatment virtually had no effect on spermatogenesis. A dose of 19 mg/kg slightly suppressed spermatogenesis 60 days after treatment, and severe suppression occurred after another 70 days of dosing. Spermatogenesis was completely recovered at the end of the recovery phase. Large dose (625 mg/kg) TU treatment did not significantly affect spermatogenesis and was well tolerated by animals. Conclusion: TU injection reversibly suppresses spermatogenesis in rats.
