DNA strand breaks in ejaculated human spermatozoa: comparison of susceptibility to the nick translation and terminal transferase assays.
The nick translation and terminal transferase assays have been compared to test their relative efficiency in detecting DNA breakage in ejaculated human spermatozoa. The results have been correlated with the percentage of chromomycin A3 positive sperm, a fluorochrome that is indicative of the protamination state of sperm. Examination of the ejaculated sperm of 30 subjects revealed that the percentage of positivity to the nick translation and terminal transferase assays did not differ, even when using different fixatives. It is concluded that the inability of the two assays to distinguish the type of DNA damage, as is possible in somatic nuclei, is most probably linked to the unique nature of sperm chromatin. It is proposed that the presence of the damaged DNA may be the remnants of an imperfect spermiogenesis, probably related to an inadequate protamine deposition. This is supported by the strong correlation between the presence of DNA damage and underprotamination as evidenced by chromomycin A3.
Manicardi
G C
GC
Department of Animal Biology, University of Modena, Italy.
Tombacco
A
A
Bizzaro
D
D
Bianchi
U
U
Bianchi
P G
PG
Sakkas
D
D
D014128:Chromomycin A3; D004249:DNA Damage; D004542:Ejaculation; D005456:Fluorescent Dyes; D005821:Genetic Techniques; D006801:Humans; D007248:Infertility, Male / Q000473:pathology*; D008297:Male; D013094:Spermatozoa / Q000473:pathology*
D005456:Fluorescent Dyes; D014128:Chromomycin A3