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Sexing river buffalo ( Bubalus bubalis L.), sheep ( Ovis aries L.), goat ( Capra hircus L.), and cattle spermatozoa by double color FISH using bovine ( Bos taurus L.) X‐ and Y‐painting probes
Author(s) -
Di Berardino Dino,
Vozdova Miluse,
Kubickova Svatava,
Cernohorska Halina,
Coppola Giuseppe,
Coppola Gianfranco,
Enne Giuseppe,
Rubes Jiri
Publication year - 2004
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.20016
Subject(s) - sexing , biology , capra hircus , sperm , bubalus , semen , ovis , andrology , bovidae , y chromosome , ploidy , water buffalo , zoology , veterinary medicine , anatomy , genetics , ecology , gene , medicine
River buffalo, sheep, and goat spermatozoa were cross‐hybridized using double color fluorescence in situ hybridization (FISH) with bovine Xcen‐ and Y‐chromosome painting probes, prepared by DOP‐PCR of laser‐microdissected‐catapulted chromosomes, to investigate the possibility of using bovine probes for sexing sperm of other members of the family Bovidae. Before sperm analysis, the probes were hybridized on metaphase chromosomes of each species, as control. Frozen‐thawed spermatozoa of cattle, river buffalo, sheep, and goat were decondensed in suspension with 5 mM DTT. Sperm samples obtained from three individuals of each species were investigated, more than 1,000 spermatozoa were scored in each animal. FISH analysis of more than 12,000 sperm revealed high level of sperm with X‐ or Y‐signals in all of the species investigated, indicating FISH efficiency over 99%. Significant interspecific differences were detected in the frequency of aberrant spermatozoa (aneuploid and diploid) between goat (0.393%) and sheep (0.033%) ( P < 0.01), goat and cattle (0.096%) ( P < 0.5), as well as between river buffalo (0.224%) and sheep ( P < 0.5). There was no significant difference between river buffalo and cattle. The present study demonstrated that it is possible to use bovine X‐Y painting probes for sexing and analyzing sperm of other species of the family, thus facilitating future studies on the incidence of chromosome abnormalities in sperm as well as on sex predetermination of embryos for the livestock industry. Mol. Reprod. Dev. 67: 108–115, 2004. © 2004 Wiley‐Liss, Inc.