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タイトル: Sexing of Dog Sperm by Fluorescence In Situ Hybridization
著者: OI, Maya
YAMADA, Keisuke
HAYAKAWA, Hiroyuki
SUZUKI, Hiroshi
鈴木, 宏志
キーワード: Dog
Fluorescence in situ hybridization (FISH)
Sexing
Sperm
発行日: 2013
出版者: 日本繁殖生物学会
誌名: Journal of Reproduction and Development
巻: 59
号: 1
開始ページ: 92-96
抄録: Effective preselection of sex has been accomplished in several species of livestock and also in humans using the flow cytometric sperm sorting method. A guaranteed high sorting accuracy is a key prerequisite for the widespread use of sperm sexing. The standard validation method is flow cytometric remeasurement of the DNA content of the sexed sperm. Since this method relies on the same instrument that produced the original sperm separation, it is not truly independent. Therefore, to be able to specifically produce either male or female offspring in the dog, we developed a method of direct visualization of sex chromosomes in a single sperm using fluorescence in situ hybridization (FISH) as a validation method. Denaturation of canine spermatozoa by immersion in 1 M NaOH for 4 min yielded consistent hybridization results with over 97% hybridization efficiency and a good preservation of sperm morphology. There was no significant difference between the theoretical ratio (50:50) and the observed ratio of X- and Y-chromosome-bearing spermatozoa in any of the three dogs. In addition, the mean purities of flow-sorted sex chromosomes in spermatozoa of the three dogs were 90.8% for the X chromosome fraction and 89.6% for the Y chromosome fraction. This sorting was evaluated by using the dual color FISH protocol. Therefore, our results demonstrated that the FISH protocol worked reliably for both unsorted and sexed sperm samples.
URI: http://ir.obihiro.ac.jp/dspace/handle/10322/3858
ISSN: 1348-4400
0916-8818
出現コレクション:05001学術雑誌論文

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