Specific cytogenetic labeling of bovine spermatozoa bearing X or Y chromosomes using fluorescent in situ hybridization (FISH).
X and Y specific probes were identified in order to apply the fluorescent in situ hybridization (FISH) technique to bovine spermatozoa. For Y chromosome detection, the BRY4a repetitive probe, covering three quarters of the chromosome, was used. For X chromosome detection, a goat Bacterial Artificial Chromosome (BAC) specific to the X chromosome of bovine and goats and giving a strong FISH signal was used. Each probe labeled roughly 45% of sperm cells. The hybridization method will be useful for evaluating the ratio of X- and Y- bearing spermatozoa in a sperm sample and consequently can be used to evaluate the efficiency of sperm sorting by different techniques such as flow cytometry.
UNCEIA, Services techniques, Maisons-Alfort, France. piumi@biotec.jouy.inra.fr
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F
Piumi
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D
Vaiman
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EP
Cribiu
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B
Guérin
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P
Humblot
D000818:Animals; D002417:Cattle / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence / Q000662:veterinary; D008297:Male; D016133:Polymerase Chain Reaction; D012744:Sex Ratio; D013094:Spermatozoa; D014960:X Chromosome / Q000737:chemistry; D014998:Y Chromosome / Q000737:chemistry
D015342:DNA Probes