FISH assessment of aneuploidy frequencies in mature and immature human spermatozoa classified by the absence or presence of cytoplasmic retention.
Previously, a relationship has been found between diminished cellular maturity of human spermatozoa and low-level expression of the testis-specific chaperone protein, HspA2. Because HspA2 is a component of the synaptonemal complex in rodents, and assuming that this is also the case in men, it was postulated that the frequency of chromosomal aneuploidies would be higher in immature versus mature spermatozoa. This question was examined in spermatozoa from semen and from 80% Percoll pellets (enriched for mature spermatozoa) of the same ejaculate in 10 oligozoospermic men. Immature spermatozoa with retained cytoplasm, which signifies spermiogenetic arrest, were identified by immunocytochemistry. Using fluorescence in-situ hybridization (FISH), approximately 7000 sperm nuclei were evaluated in each of the 20 fractions (142 086 spermatozoa in all) using centromeric probes for the X, Y and 17 chromosomes. The proportions of immature spermatozoa were 45.4 +/- 3.4 versus 26.6 +/- 2.2% in the two semen versus the Percoll groups (medians: 48.2 versus 25%, P < 0.001, n = 300 spermatozoa per fraction, total 6000 spermatozoa). There was also a concomitant decline in total disomy, total diploidy and total aneuploidy frequencies in the 80% Percoll versus semen fractions (0.17 versus 0.54%, 0.14 versus 0.26% and 0.31 versus 0.81% respectively, P < 0.001 in all comparisons). The mean decline of aneuploidies was 2.7-fold. With regard to the hypothesis that aneuploidies are related to sperm immaturity, there was a close correlation between the incidence of immature spermatozoa and disomies (r = 0.7, P < 0.001) but no correlation with diploidies (r = 0.03), indicating that disomies originate primarily in immature spermatozoa. It is suggested that the common factor underlying sperm immaturity and aneuploidies is the diminished expression of HspA2. In addition, the lack of this chaperone may also cause diminished cellular transport of proteins, such as DNA-repair enzymes or of the retention of cytoplasm that is extruded from normally maturing spermatozoa during spermiogenesis.
Kovanci
E
E
The Sperm Physiology Laboratory, Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, CT, USA.
Kovacs
T
T
Moretti
E
E
Vigue
L
L
Bray-Ward
P
P
Ward
D C
DC
Huszar
G
G
D000782:Aneuploidy; D002467:Cell Nucleus / Q000648:ultrastructure; D002469:Cell Separation; D016922:Cellular Senescence; D002499:Centrifugation, Density Gradient; D003402:Creatine Kinase / Q000032:analysis; D003593:Cytoplasm / Q000648:ultrastructure*; D015342:DNA Probes; D004171:Diploidy; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D007527:Isoenzymes / Q000032:analysis; D008297:Male; D009845:Oligospermia / Q000473:pathology; D012661:Semen / Q000166:cytology; D013094:Spermatozoa / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure*; D014960:X Chromosome; D014998:Y Chromosome
D015342:DNA Probes; D018840:HSP70 Heat-Shock Proteins; C091879:HSPA2 protein, human; D007527:Isoenzymes; D003402:Creatine Kinase