Detection of water buffalo sex chromosomes in spermatozoa by fluorescence in situ hybridization.
In order to identify X- and Y-bearing spermatozoa in water buffalo by fluorescence in situ hybridization (FISH), some available probes of closely related species were examined. An X- and Y-specific probe set, made from flow sorted yak chromosomes, labelled in somatic metaphases of water buffalo the whole X and Y, respectively, except their centromere regions. A cattle Y-chromosome repeat sequence (BC1.2) showed strong signal on the telomere region of the buffalo Y-chromosome, demonstrating the evolutionary conservation of this locus in water buffalo. In hybridization experiments with spermatozoa from five buffaloes, the yak X-Y paint set demonstrated clear signals in more than 92% (46.8% X and 45.8% Y) of the cells. Using the cattle Y-chromosome specific BC1.2 probe, clear hybridization signal was detected in more than 48% of the cells. Statistical analysis showed that there was no significant difference between bulls or from the expected 50 : 50 ratio of X- and Y-bearing cells. The probes presented here are reliable to assess separation of X- and Y-bearing spermatozoa.
Révay
T
T
Budapest University of Technology and Economics, Department of Biochemistry and Food Technology, Budapest, Megyetem rkp. 3. Hungary.
Kovács
A
A
Presicce
G A
GA
Rens
W
W
Gustavsson
I
I
D000818:Animals; D002020:Buffaloes; D015342:DNA Probes; D017404:In Situ Hybridization, Fluorescence / Q000662:veterinary*; D008297:Male; D012730:Sex Chromosomes / Q000737:chemistry*; D013094:Spermatozoa / Q000737:chemistry*
D015342:DNA Probes