精子携带的HBV DNA在小鼠早期胚胎中的复制与表达

doi:10.3969/j.issn.1004-616x.2005.03.013

癌变·畸变·突变 ›› 2005, Vol. 17 ›› Issue (3): 175-178.doi: 10.3969/j.issn.1004-616x.2005.03.013

精子携带的HBV DNA在小鼠早期胚胎中的复制与表达

熊小芳;黄天华;谢庆东;王晓梅;陈桂兰

汕头大学医学院生殖医学研究中心，广东　汕头　515041

收稿日期:2004-11-30 修回日期:2005-02-28 出版日期:2005-05-30 发布日期:2005-05-30
通讯作者: 黄天华

Replication and Expression of Sperm-mediated HBV DNA in Early Mouse Embryo

XIONG Xiao-fang;HUANG Tian-hua;XIE Qing-dong;WANG Xiao-mei;CHEN Gui-lan

Research Center for Reproductive Medicine, Shantou University Medical College, Shantou 515041,Guangdong,China

Received:2004-11-30 Revised:2005-02-28 Online:2005-05-30 Published:2005-05-30
Contact: HUANG Tian-hua

摘要/Abstract

摘要： 背景与目的：研究乙型肝炎病毒(Hepatitis B virus,HBV) DNA在早期胚胎中能否复制与表达，探讨HBV经雄性生殖细胞垂直传递的可能性。材料与方法：成熟雄鼠麻醉后双侧睾丸注射经脂质体DOSPER包裹的HBV质粒，手术后雄鼠与超排雌鼠合笼交配，用荧光原位杂变(Fluorescence in situ hybridization,FISH)分别检测单细胞胚和二细胞胚间期核中HBV DNA的存在与复制，用逆转录聚含酶链反应(Reverse transcriptase-polymerase chain reaction,RT-PCR)和免疫荧光方法检测HBV基因在二细胞胚胎中的表达。结果：在单细胞胚和二细胞胚间期核内发现HBV DNA阳性杂交信号，RT-PCR可以扩增出HBx基因的特异条带，免疫荧光也可见到HBsAg阳性信号。结论： HBV DNA进入雄性生殖细胞后，可以整合到宿主基因组内，通过自然受精随精子进入卵母细胞，并在早期胚胎中复制与表达。提示HBV有可能通过雄性生殖细胞进行垂直传递。

关键词: 乙型肝炎病毒DNA, 复制与表达, 早期胚胎, 垂直传递

Abstract: BACKGROUND ＆ AIM: To explore the feasibility of hepatitis B virus (HBV) vertical transmission via male germ line, the replication and expression of sperm-mediated HBV DNA in mouse embryo were studied. MATERIAL AND METHODS: Young adult male mice were injected with HBV DNA-liposome complex. These males were serially mated with superovulated females 2 days after injection. Fluorescence in situ hybridization (FISH) was carried out to confirm the integration of HBV DNA into male pronucleus and its replication with cell division in embryonic development.(Reverse transcriptase polymerase chain reaction,RT-PCR) and immunofluoresence assay were performed to observe the expression of the HBV gene in two-cell stage. RESULTS: FISH demonstrated that the male pronuclei in some one-cell embryos and the each nucleus in some two-cell embryos presented positive signals. RT-PCR showed the specific bands of cDNA of HBx DNA in some two-cell embryos. Immunofluorescence assay presented the positive signals for HBsAg in some two-cell embryos. CONCLUSION: Our results provided the direct evidence for that HBV DNA are able to transmit vertically to next generation via male germ line.

Key words: hepatitis B virus DNA, replication and expression, early embryo, vertical transmission

熊小芳, 黄天华, 谢庆东, 王晓梅, 陈桂兰. 精子携带的HBV DNA在小鼠早期胚胎中的复制与表达[J]. 癌变·畸变·突变, 2005, 17(3): 175-178.

XIONG Xiao-fang, HUANG Tian-hua, XIE Qing-dong, WANG Xiao-mei, CHEN Gui-lan. Replication and Expression of Sperm-mediated HBV DNA in Early Mouse Embryo[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2005, 17(3): 175-178.

[1]	程琳, 孙平楠, 谢庆东, 周小玲. 预扩增qPCR方法检测少量小鼠早期胚胎细胞中DNA甲基化相关基因的表达[J]. 癌变·畸变·突变, 2016, 28(1): 51-55.
[2]	周莉,王永,王蓉,许丽,骆永伟,王华,孙祖越. 生育力与早期胚胎发育毒性试验中雄性大鼠阳性模型的建立[J]. 癌变·畸变·突变, 2013, 25(4): 311-315.
[3]	高郁森, 谢庆东. HIV-1 gag基因经卵母细胞垂直传递模型的建立[J]. 癌变·畸变·突变, 2010, 22(1): 39-0042.
[4]	倪永林(综述), 林少达(审校). Bcl_2蛋白家族对卵泡和早期胚胎生长发育的调节[J]. 癌变·畸变·突变, 2008, 20(2): 166-168.
[5]	徐珊珊, 黄天华, 谢庆东, 吴丛梅, 吴德生. pIRES2-EGFP-DAZ3质粒构建及金黄地鼠2-细胞胚中人DAZ基因的复制与表达[J]. 癌变·畸变·突变, 2006, 18(3): 180-184.
[6]	黄天华;谢庆东;陈霓彤. 精子携带的乙肝病毒基因在金黄地鼠胚胎中的复制与表达[J]. , 2005, 17(4): 193-197.
[7]	陈桂兰;黄天华;谢庆东;王晓梅;熊小芳. 卵母细胞带的HBV NA在小鼠期胚胎中的制与表达[J]. , 2005, 17(3): 179-182.

精子携带的HBV DNA在小鼠早期胚胎中的复制与表达

Replication and Expression of Sperm-mediated HBV DNA in Early Mouse Embryo

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 7

编辑推荐

Metrics

本文评价