荧光原位杂交(FISH) 检测人精子染色体非整倍体率方法的研究

doi:10.3969/j.issn.1004-616x.1998.05.001

癌变·畸变·突变 ›› 1998, Vol. 10 ›› Issue (5): 267-271.doi: 10.3969/j.issn.1004-616x.1998.05.001

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荧光原位杂交(FISH) 检测人精子染色体非整倍体率方法的研究

郑履康　邓丽霞　张　桥

中山医科大学公共卫生学院遗传毒理研究室　广州　510089

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1998-09-30 发布日期:1998-09-30

STUDY ON THE ANEUPLOIDY FREQUENCY IN HUMAN SPERM CHROMOSOME USING FLUORESCENCE IN SITU HYBRIDIZATION ( FISH)

Zheng Lukang , Deng Lixia , Zhang qiao

L ab. of Genetic Toxicology , School of Public Heal th , S un Yat2Sen University of Medical Sciences , Guangz hou 510089

Received:1900-01-01 Revised:1900-01-01 Online:1998-09-30 Published:1998-09-30

摘要/Abstract

摘要： 本研究建立了荧光原位杂交(FISH) 方法,并测定了十名正常健康男子精子X 染色体非整倍体率。取少量精液经洗后制片,用二梳苏糖醇(DTT) 和二碘水杨酸锂(L IS) 处理,使精子头部紧密的染色质去凝集,然后与生物素标记的α- 卫星X 染色体特异DNA 探针(DXZ1) 作原位杂交;用Cy3 - 链亲和素检测杂交信号。结果在荧光显微镜下可清楚见到带有红色杂交点信号的X 精子。杂交率平均为48.62 % ( 47.54 % -49.36 %) 。此值与X 精子占50 %的理论值差别无显著性。总共计数55453 个X 精子,X 染色体双体率为0.06 %;此结果与国外一些学者先前报导的结果一致,本研究提供的正常人X 精子双体率的资料,为今后研究职业或环境潜在非整倍体诱导剂对人精子的影响提供有价值的材料。

关键词: 荧光原位杂交, 人精子染色体, 非整倍体率

Abstract: The f requency of 　Human sperm chromosome aneuploidy was assessed from 10 normal and health adult s by using the method of FISH. Sperm heads were decondensed with dithilthreitol (DTT) and lithium diiodosalicylate (LIS) , then the sperm nuclei were hybridized with biotin labeled alpha satellite X chromosome specific DNA probe (DXZ1) in 37 ℃ overnight . The hybridization signals were detected with Cy32St reptavidin. The red hybridization signals in X sperms were clearly visible under the Nikon fluorescence microscope. The hybridization efficiency was 48. 62 % ( rance 47. 54 % to 49. 36 %) . A total 55453 X sperm nuclei were observed. The frequency of disomic sperms was 0. 06 %. There was no significant difference observed in disomic frequency of X sperms between ours and that reported by former authors. The information provided by this paper is valuable for further studies on men exposed occupationally or environmentally to potential aneuploidogenic agents.

Key words: Fluorescence in situ hybridization (FISH), Human sperm chromosome, Aneuploidy frequency

郑履康　邓丽霞　张　桥. 荧光原位杂交(FISH) 检测人精子染色体非整倍体率方法的研究[J]. 癌变·畸变·突变, 1998, 10(5): 267-271.

Zheng Lukang , Deng Lixia , Zhang qiao. STUDY ON THE ANEUPLOIDY FREQUENCY IN HUMAN SPERM CHROMOSOME USING FLUORESCENCE IN SITU HYBRIDIZATION ( FISH)[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 1998, 10(5): 267-271.

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荧光原位杂交(FISH) 检测人精子染色体非整倍体率方法的研究

STUDY ON THE ANEUPLOIDY FREQUENCY IN HUMAN SPERM CHROMOSOME USING FLUORESCENCE IN SITU HYBRIDIZATION ( FISH)

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