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Rapid and simultaneous detection of chromosome Y‐ and 1‐bearing porcine spermatozoa by fluorescence in situ hybridization
Author(s) -
Kawarasaki Tatsuo,
Sone Masaru,
Yoshida Mitsutoshi,
Bamba Kimio
Publication year - 1996
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/(sici)1098-2795(199604)43:4<548::aid-mrd18>3.0.co;2-v
Subject(s) - biology , in situ hybridization , fluorescence in situ hybridization , in situ , chromosome , y chromosome , microbiology and biotechnology , bearing (navigation) , fluorescence , genetics , gene , gene expression , physics , cartography , quantum mechanics , meteorology , geography
This study was carried out to develop a rapid and simultaneous detection system of chromosome Y‐ and 1‐bearing porcine spermatozoa by fluorescence in situ hybridization (FISH). Chromosome Y‐ and 1‐specific DNA probes were produced by polymerase chain reaction with digoxigenin (Dig)‐ or biotin‐dUTP. The hybridization probe mixture of labeled Y‐chromosome and chromosome 1‐specific DNA was applied to the preparation, immediately denatured at 75°C for 8 min, hybridized for 5 min at 37°C and overall FISH steps were done within a few hours. When double FISH with Dig‐labeled chromosome Y‐specific and biotin‐labeled chromosome 1‐specific probes was applied to sperm nuclei pretreated with dithiothreitol, the average of 50.9% of sperm nuclei had the Dig‐signal, 99.2% of the sperm nuclei had the biotin‐signal and the average of 0.3% of sperm nuclei showed no signal. The putative rate of Y‐bearing spermatozoa ranged from 49.8% to 52.8% among 5 boars and the average putative rate of Y‐bearing spermatozoa was 51.0%. The results indicated that a rapid and simultaneous FISH with chromosome Y‐ and 1‐specific porcine DNA probes produced by PCR made possible more accurate assessment of Y‐bearing porcine spermatozoa. © 1996 Wiley‐Liss, Inc.