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 ch...

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Bibliographic Details
Published in:Molecular human reproduction 1998, Vol.4 (1), p.61-66
Main Authors: WEISSENBERG, R, AVIRAM, A, GOLAN, R, LEWIN, L. M, LEVRON, J, MADGAR, I, DOR, J, BARKAI, G, GOLDMAN, B
Format: Article
Language:English
Subjects:
Adult
Aneuploidy
Biological and medical sciences
Birth control
Chromatin
Chromosomes, Human, Pair 18 - genetics
Flow Cytometry - methods
Gynecology. Andrology. Obstetrics
Humans
In Situ Hybridization, Fluorescence - methods
Male
Medical sciences
Meiosis
Nondisjunction, Genetic
Oligospermia - pathology
Spermatozoa - abnormalities
Spermatozoa - cytology
Spermatozoa - physiology
Sterility. Assisted procreation
X Chromosome - genetics
Y Chromosome - genetics
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Summary: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.
ISSN:1360-9947
1460-2407
1460-2407
DOI:10.1093/molehr/4.1.61