NDLI: Sexing of Dog Sperm by Fluorescence In Situ Hybridization

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Sexing of Dog Sperm by Fluorescence In Situ Hybridization

Content Provider	Semantic Scholar
Author	Oi, Maya Yamada, Keisuke Hayakawa, Hiroyuki Suzuki, Hiroshi
Copyright Year	2013
Abstract	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.
Starting Page	92
Ending Page	96
Page Count	5
File Format	PDF HTM / HTML
Alternate Webpage(s)	https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/28/aa/jrd-59-092.PMC3943228.pdf
PubMed reference number	23059640v1
Volume Number	59
Journal	The Journal of reproduction and development
Language	English
Access Restriction	Open
Subject Keyword	Biologic Preservation Canis familiaris Chromosomes Congenital chromosomal disease Crossbreeding Dual Fluorescence Fluorescent in Situ Hybridization Nucleic Acid Hybridization Sex Factors Spermatozoa Cell Count
Content Type	Text
Resource Type	Article

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