DETECTION OF ANEUPLOID HUMAN SPERM BY FLUORESCENCE INSITU HYBRIDIZATION - EVIDENCE FOR A DONOR DIFFERENCE IN FREQUENCY OF SPERM DISOMIC FORCHROMOSOMES-I AND CHROMOSOMES-Y

Fluorescence in situ hybridization with repetitive-sequence DNA probes was used to detect human sperm disomic for chromosomes 1 and Y in thr ee healthy men. Data on these same men had been obtained previously, u sing the human-sperm/hamster-egg cytogenetic technique, providing a cy togenetic reference for validating sperm hybridization measurements. A ir-dried smears were prepared from semen samples and treated with DTT and lithium diiodosalicylate to expand sperm chromatin. Hybridization with fluorescently tagged DNA probes for chromosomes 1 (pUC177) or Y ( pY3.4) yielded average frequencies of sperm with two fluorescent domai ns of 14.2+/-2.4/10,000 and 5.6+/-1.6/10,000 sperm, respectively. Thes e frequencies did not differ statistically from frequencies of hyperpl oidy observed for these chromosomes with the hamster technique. In add ition, frequencies of disomic sperm from one donor were elevated appro ximately 2.5-fold above those of other donors, for both chromosomes 1 (P = .045) and Y (P = .01), consistent with a trend found with the ham ster technique. We conclude that fluorescence in situ hybridization to sperm chromosomes provides a valid and promising measure of the frequ ency of disomic human sperm.