Aim： To investigate whether adriamycin induces DNA damage and the formation of γH2AX （the phosphorylated form of histone H2AX） foci in mature spermatozoa. Methods： Human spermatozoa were treated with adriamycin a...Aim： To investigate whether adriamycin induces DNA damage and the formation of γH2AX （the phosphorylated form of histone H2AX） foci in mature spermatozoa. Methods： Human spermatozoa were treated with adriamycin at different concentrations, γH2AX was analyzed by immunofluorescent staining and flow cytometry and doublestrand breaks （DSB） were detected by the comet assay. Results： The neutral comet assay revealed that the treatment with adriamycin at 2 μg/mL for different times （0.5, 2, 8 and 24 h）, or for 8 h at different concentrations （0,4, 2 and 10 μg/mL）, induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of γH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP 1 with γH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase （PI3K） family, abolished the co-appearance of these two proteins with γH2AX. Conclusion： Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/ repair proteins as somatic cells.展开更多
Chromosomal abnormalities and Y chromosome microdeletions are considered to be the two more common genetic causes of spermatogenic failure.However,the relati on ship between chromosomal aberrations and Y chromosome mi...Chromosomal abnormalities and Y chromosome microdeletions are considered to be the two more common genetic causes of spermatogenic failure.However,the relati on ship between chromosomal aberrations and Y chromosome microdeletio ns is still un clear.This study was to investigate the incidenee and characteristics of chromosomal aberrations and Y chromosome microdeletions in infertile men,and to explore whether there was a correlation between the two genetic defects of spermatogenic failure.A 7-year retrospective study was conducted on 5465 infertile men with nonobstructive azoospermia or oligozoospermia.Karyotype analysis of peripheral blood lymphocytes was performed by standard G-banding techniques.Y chromosome microdeletions were screened by multiplex PCR amplification with six specific sequence-tagged site(STS)markers.Among the 5465 infertile men analyzed,371(6.8%)had Y chromosome microdeletions and the prevalence of microdeletions in azoospermia was 10.5%(259/2474)and in severe oligozoospermia was 6.3%(107/1705).A total of 4003(73.2%)infertile men underwent karyotyping;370(9.2%)had chromosomal abnormalities and 222(5.5%)had chromosomal polymorphisms.Karyotype analysis was performed on 272(73.3%)patients with Y chromosome microdeletions and 77(28.3%)had chromosomal aberrations,all of which involved sex chromosomes but not autosomes.There was a sign ifica nt d iff ere nee in the frequency of chromosomal abno rmalities betwee n men with and without Y chromosome microdeletions(P<0.05).展开更多
文摘Aim： To investigate whether adriamycin induces DNA damage and the formation of γH2AX （the phosphorylated form of histone H2AX） foci in mature spermatozoa. Methods： Human spermatozoa were treated with adriamycin at different concentrations, γH2AX was analyzed by immunofluorescent staining and flow cytometry and doublestrand breaks （DSB） were detected by the comet assay. Results： The neutral comet assay revealed that the treatment with adriamycin at 2 μg/mL for different times （0.5, 2, 8 and 24 h）, or for 8 h at different concentrations （0,4, 2 and 10 μg/mL）, induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of γH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP 1 with γH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase （PI3K） family, abolished the co-appearance of these two proteins with γH2AX. Conclusion： Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/ repair proteins as somatic cells.
基金the National Key Research and Development Program of China(No.2016YFC1000703)the Medicine and Health Science and Technology Plan Projects in Zhejiang Province(No.2014KYA246)the National Natural Science Foundation of China(Nos.81801441 and 81300532)
基金Project supported by the National Natural Science Foundation of China(Nos.81370725 and 81370726)the Natural Science Foundation of Zhejiang Province(No.LQ14H040004)the Key Discipline of Obstetrics of Zhejiang Province,China
基金We should like to thank our patients for agreeing to donate their personal data and allowing the data to be published.We are grateful to Dr.Jiong Gao(BGI Genomics,BGI-Shenzhen,Shenzhen,China)for improving the article.The study was funded by the National Key Research and Development Program of China(Grant No.2018YFC1004903,and 2016YFC1000703)Key Research and Development Program of Zhejiang Province(Grant No.2019C03025)+1 种基金the National Nature Science Foundation of China(Grant No.81801441)and Zhejiang Provincial Natural Science Foundation of China(Grant No.LQ19H090019).
文摘Chromosomal abnormalities and Y chromosome microdeletions are considered to be the two more common genetic causes of spermatogenic failure.However,the relati on ship between chromosomal aberrations and Y chromosome microdeletio ns is still un clear.This study was to investigate the incidenee and characteristics of chromosomal aberrations and Y chromosome microdeletions in infertile men,and to explore whether there was a correlation between the two genetic defects of spermatogenic failure.A 7-year retrospective study was conducted on 5465 infertile men with nonobstructive azoospermia or oligozoospermia.Karyotype analysis of peripheral blood lymphocytes was performed by standard G-banding techniques.Y chromosome microdeletions were screened by multiplex PCR amplification with six specific sequence-tagged site(STS)markers.Among the 5465 infertile men analyzed,371(6.8%)had Y chromosome microdeletions and the prevalence of microdeletions in azoospermia was 10.5%(259/2474)and in severe oligozoospermia was 6.3%(107/1705).A total of 4003(73.2%)infertile men underwent karyotyping;370(9.2%)had chromosomal abnormalities and 222(5.5%)had chromosomal polymorphisms.Karyotype analysis was performed on 272(73.3%)patients with Y chromosome microdeletions and 77(28.3%)had chromosomal aberrations,all of which involved sex chromosomes but not autosomes.There was a sign ifica nt d iff ere nee in the frequency of chromosomal abno rmalities betwee n men with and without Y chromosome microdeletions(P<0.05).
