Sequential G-banding FISH on human sperm chromosomes.
A method that allows the performance of double-colour chromosome painting (FISH) on previously G-banded human sperm metaphases has been developed. Sperm chromosomes were obtained by using the fusion technique between zona-free hamster oocytes and human spermatozoa. Single- and double-colour chromosome painting was performed using DNA libraries specific for chromosomes X, Y and 21 on either unstained or G-banded preparations. The hybridization efficiency was very high (98%). The sequential staining technique is very useful for analyses of structural (stable) and numerical chromosome aberrations in human sperm and thus can increase the efficiency of the human sperm-hamster oocytes fusion system to assess the risk to human germ cells as a result of endogenous and exogenous factors.
Colls
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Departament de Biologia Cellular i Fisiologia, Facultat de Medicina, Universitat Autonoma de Barcelona, Bellaterra, Spain.
Templado
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Martínez-Pasarell
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Darroudi
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F
Natarajan
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AT
D000328:Adult; D000818:Animals; D002871:Chromosome Banding; D002891:Chromosomes, Human, Pair 21 / Q000737:chemistry / Q000235:genetics; D006224:Cricetinae; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007211:Indoles; D008297:Male; D008677:Metaphase / Q000235:genetics; D012235:Rhodamines; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology*; D013194:Staining and Labeling; D014960:X Chromosome / Q000737:chemistry / Q000235:genetics; D014998:Y Chromosome / Q000737:chemistry / Q000235:genetics
D005456:Fluorescent Dyes; D007211:Indoles; D012235:Rhodamines; C009434:tetramethylrhodamine isothiocyanate; C007293:DAPI; D016650:Fluorescein-5-isothiocyanate