Free Access
Genet. Sel. Evol.
Volume 33, Number 1, January-February 2001
Page(s) 89 - 98
DOI: 10.1051/gse:2001111

Genet. Sel. Evol. 33 (2001) 89-98

Specific cytogenetic labeling of bovine spermatozoa bearing X or Y chromosomes using fluorescent in situ hybridization (FISH)

François Piumia, Daniel Vaimanb, Edmond-Paul Cribiub, Bernard Guérina and Patrice Humblota

a  UNCEIA, Services techniques, Maisons-Alfort, France
b  Département de génétique animale, Laboratoire de génétique biochimique et de cytogénétique, Groupe petits ruminants, Institut national de la recherche agronomique, 78352 Jouy-en-Josas, France

(Received 14 August 2000; accepted 24 October 2000)

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.

Key words: fluorescent in situ hybridization / spermatozoon / X chromosome / Y chromosome / bacterial artificial chromosome

Correspondence and reprints: F. Piumi

© INRA, EDP Sciences 2001