Infertility is a severe problem that affects 10%-15% of couples worldwide, among which male infertility is a factor in ~50% of the cases [1]. Azoospermia, which is defined as the absence of sperm in at least two ejacu...Infertility is a severe problem that affects 10%-15% of couples worldwide, among which male infertility is a factor in ~50% of the cases [1]. Azoospermia, which is defined as the absence of sperm in at least two ejaculates, can be divided into two types: obstructive azoospermia (OA) and non-OA (NOA)[1]. OA patients have normal spermatogenesis and mature sperm in testes and can father a biological child through testicular sperm retrieval and intracytoplasmic sperm injection (ICSI)[2]. By contrast, NOA patients usually have various degrees of spermatogenesis impairment and can be faced with complicated assisted reproductive technology (ART) strategies to father a biological child [2]. The precise diagnosis of spermatogenesis in NOA patients is crucial for clinical decision making regarding ART.展开更多
Mammalian spermatogenesis is maintained by a rare population of spermatogonial stem cells(SSCs),which are important for male fertility. SSCs remain a subset of undifferentiated spermatogonia, which can be isolated by ...Mammalian spermatogenesis is maintained by a rare population of spermatogonial stem cells(SSCs),which are important for male fertility. SSCs remain a subset of undifferentiated spermatogonia, which can be isolated by a combination of surface markers. Specific markers to identify and isolate undifferentiated spermatogonia are lacking. Ussp1, a transcript previously annotated as long noncoding RNA(RIKEN cDNA 4933427D06, Gene ID: 232217), virtually encodes a membrane protein, USSP1, in a highly testisspecific manner in mouse. We demonstrate its expression on the membrane of undifferentiated spermatogonia by a homemade polyclonal rabbit antibody against the protein. In vivo, USSP1~+ clusters consist mainly of As, Apr(GFRa1~+) and Aal(PLZF~+) cells. USSP1~+ cells exhibit enrichment of undifferentiated spermatogonia, as shown by increased expression of SSC self-renewal molecular markers and the potential to form SSC clones in vitro and in vivo. However, Ussp1 knockout did not affect the number of SSCs or spermatogenesis in mice. Thy1~+ cells from Ussp1 null mice did not show any defect in the SSC colony formation capacity, indicating that USSP1 is not essential for SSC self-renewal. Our data demonstrate that Ussp1 is specifically expressed in undifferentiated murine spermatogonia, indicating the potential to sort undifferentiated spermatogonia with USSP1 antibodies. Ussp1 might be a good maker for SSC enrichment in neonatal mice.展开更多
In this study,three cDNA sequences corresponding to cytochrome P450C17(CYP17 I),3β-hydroxysteroid dehydrogenase(3β-HSD)and androgen receptor(AR)were isolated from spotted sea bass(Lateolabrax maculatus).The mRNA abu...In this study,three cDNA sequences corresponding to cytochrome P450C17(CYP17 I),3β-hydroxysteroid dehydrogenase(3β-HSD)and androgen receptor(AR)were isolated from spotted sea bass(Lateolabrax maculatus).The mRNA abundances of CYP17 I and 3β-HSD increased from stageⅡto stageⅤwith a significant increase at stageⅤ,and the highest abundance of AR mRNA was detected at stageⅢin testicular development cycle.CYP17 I,3β-HSD and AR transcripts were obviously abundant in steroidogenesis tissues such as testis,brain,head kidney among others.Strong and positive signals were observed mainly in interstitial cell regions of L.maculatus testis as were measured with in situ hybridization method.Significant increases of CYP17 I and 3β-HSD transcripts were detected after 12–48 h hCG(human chorionic gonadotropin)and GnRHa(gonadotropin-releasing hormone analogue)treatments.However,an opposite relationship was found for AR in testis at the same time.In addition,decreasing trends of CYP17 I and 3β-HSD mRNA were observed in testis of L.maculatus in freshwater group(FW)from day 2 to day 6,and mRNA abundance of AR increased in brackish water(BW)group from day 4 to day 8.These findings revealed that these three steroid synthesis genes are import for testicular development,hormone and salinity treatment,and provided also an insight into the mechanism of reproductive endocrine of L.maculatus.展开更多
基金This work was supported by the grants from the National Natural Science Foundation of China (No.81571486)the Open Project of Shanghai Key Laboratory of Molecular Andrology (No.SLMA-09).
文摘Infertility is a severe problem that affects 10%-15% of couples worldwide, among which male infertility is a factor in ~50% of the cases [1]. Azoospermia, which is defined as the absence of sperm in at least two ejaculates, can be divided into two types: obstructive azoospermia (OA) and non-OA (NOA)[1]. OA patients have normal spermatogenesis and mature sperm in testes and can father a biological child through testicular sperm retrieval and intracytoplasmic sperm injection (ICSI)[2]. By contrast, NOA patients usually have various degrees of spermatogenesis impairment and can be faced with complicated assisted reproductive technology (ART) strategies to father a biological child [2]. The precise diagnosis of spermatogenesis in NOA patients is crucial for clinical decision making regarding ART.
基金the National Key Research and Development Program of China(2017YFA0102801 and 2017YFC1001901)the Science and Technology Planning Project of Guangdong Province(2015B020228002)+2 种基金the National Natural Science Foundation of China(31671540)the Natural Science Foundation of Guangdong Province(2015A020212005 and 2014A030312011)the Guangzhou Science and Technology Project(201803010020).
文摘Mammalian spermatogenesis is maintained by a rare population of spermatogonial stem cells(SSCs),which are important for male fertility. SSCs remain a subset of undifferentiated spermatogonia, which can be isolated by a combination of surface markers. Specific markers to identify and isolate undifferentiated spermatogonia are lacking. Ussp1, a transcript previously annotated as long noncoding RNA(RIKEN cDNA 4933427D06, Gene ID: 232217), virtually encodes a membrane protein, USSP1, in a highly testisspecific manner in mouse. We demonstrate its expression on the membrane of undifferentiated spermatogonia by a homemade polyclonal rabbit antibody against the protein. In vivo, USSP1~+ clusters consist mainly of As, Apr(GFRa1~+) and Aal(PLZF~+) cells. USSP1~+ cells exhibit enrichment of undifferentiated spermatogonia, as shown by increased expression of SSC self-renewal molecular markers and the potential to form SSC clones in vitro and in vivo. However, Ussp1 knockout did not affect the number of SSCs or spermatogenesis in mice. Thy1~+ cells from Ussp1 null mice did not show any defect in the SSC colony formation capacity, indicating that USSP1 is not essential for SSC self-renewal. Our data demonstrate that Ussp1 is specifically expressed in undifferentiated murine spermatogonia, indicating the potential to sort undifferentiated spermatogonia with USSP1 antibodies. Ussp1 might be a good maker for SSC enrichment in neonatal mice.
基金the Chinese Agriculture Research System(No.CARS-47)Shandong Provincial Natural Science Foundation(No.ZR2016CQ21)the Key Laboratory of Mariculture(Ocean University of China),Ministry of Education(No.2018008).
文摘In this study,three cDNA sequences corresponding to cytochrome P450C17(CYP17 I),3β-hydroxysteroid dehydrogenase(3β-HSD)and androgen receptor(AR)were isolated from spotted sea bass(Lateolabrax maculatus).The mRNA abundances of CYP17 I and 3β-HSD increased from stageⅡto stageⅤwith a significant increase at stageⅤ,and the highest abundance of AR mRNA was detected at stageⅢin testicular development cycle.CYP17 I,3β-HSD and AR transcripts were obviously abundant in steroidogenesis tissues such as testis,brain,head kidney among others.Strong and positive signals were observed mainly in interstitial cell regions of L.maculatus testis as were measured with in situ hybridization method.Significant increases of CYP17 I and 3β-HSD transcripts were detected after 12–48 h hCG(human chorionic gonadotropin)and GnRHa(gonadotropin-releasing hormone analogue)treatments.However,an opposite relationship was found for AR in testis at the same time.In addition,decreasing trends of CYP17 I and 3β-HSD mRNA were observed in testis of L.maculatus in freshwater group(FW)from day 2 to day 6,and mRNA abundance of AR increased in brackish water(BW)group from day 4 to day 8.These findings revealed that these three steroid synthesis genes are import for testicular development,hormone and salinity treatment,and provided also an insight into the mechanism of reproductive endocrine of L.maculatus.