Concurrent use of flow cytometry and fluorescence in-situ hybridization techniques for detecting faulty meiosis in a human sperm sample.
Routine semen analysis in an infertile patient revealed severe teratospermia associated with malformation of head and tail in 100% of the sperm cells. Flow cytometry and fluorescence in-situ hybridization (FISH) were shown to supplement routine semen analysis by providing information on the sperm chromatin. Using flow cytometry, propidium iodide-stained spermatozoa from the same sperm sample were compared with a normal reference pool, and with human lymphocytes. The results point to a population of diploid sperm cells rather than to mature haploid spermatozoa. Numerical chromosomal abnormalities of the spermatozoa were subsequently evaluated using FISH. A total of 1000 sperm cells were scored for X and Y chromosomes, and an additional 1128 sperm cells for chromosome 18. Aneuploidy of chromosomes X and Y was revealed in 96.9% of the cells and of chromosome 18 in 90.3% of the cells. Non-disjunction of chromosome X and Y in meiosis I and II occurred in 54.8 and 2.7% of the sperm cells respectively. Non-disjunction in both meiosis I and II occurred in 39.4% of the sperm cells. A normal haploid pattern for chromosomes X and Y was observed in only 3.1%, and for chromosome 18 in 9.7%, of the cells. Using three colour FISH for the sex chromosomes and for chromosome 18, diploidy was demonstrated in 19.4% of 500 sperm cells and aneuploidy in virtually all sperm cells (99.2%). The use of flow cytometry and FISH in cases where genetic and developmental chromatin abnormalities are suspected is a valuable adjunct to other available techniques, and can guide the clinicians to decide which samples are unsuitable for intracytoplasmic injection.
Weissenberg
R
R
Andrology Unit, Sheba Medical Center, Tel Hashomer, Ramat Gan, Israel.
Aviram
A
A
Golan
R
R
Lewin
L M
LM
Levron
J
J
Madgar
I
I
Dor
J
J
Barkai
G
G
Goldman
B
B
D000328:Adult; D000782:Aneuploidy; D002843:Chromatin; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D008540:Meiosis; D009630:Nondisjunction, Genetic; D009845:Oligospermia / Q000473:pathology*; D013094:Spermatozoa / Q000002:abnormalities* / Q000166:cytology / Q000502:physiology; D014960:X Chromosome / Q000235:genetics; D014998:Y Chromosome / Q000235:genetics
D002843:Chromatin