Increased DNA damage in sperm from leukocytospermic semen samples as determined by the sperm chromatin structure assay.
OBJECTIVE
To determine DNA damage as measured by the sperm chromatin structure assay (SCSA) in subsets of human spermatozoa at different stages of maturation in patients who are undergoing infertility evaluation.


METHODS
Prospective study.


METHODS
Andrology laboratory at a tertiary care hospital.


METHODS
Fifty-six patients undergoing infertility evaluation. Patients with normal semen parameters (n = 17), abnormal semen parameters (n = 29), leukocytospermia (n = 10), and a group of healthy fertile men (n = 18) were included in the study.


METHODS
None.


METHODS
The shift of green (native DNA) to red (denatured, single-stranded DNA) fluorescence was measured and quantified using the expression alpha(t) (red fluorescence/[red + green fluorescence] per cell). Sperm DNA damage was examined in subsets of spermatozoa isolated by a three-step density gradient. The DNA damage was correlated with classic semen characteristics.


RESULTS
Leukocyte concentration in semen was directly correlated with chromatin alterations in immature and mature sperm. Leukocyte concentration in semen was also directly correlated with immature germ cell concentration and the percentage of abnormal forms in semen.


CONCLUSIONS
The increase in chromatin alterations and DNA damage in sperm, as defined by the sperm chromatin structure assay from leukocytospermic samples may be related to alterations in the regulation of spermatogenesis.
Department of Obstetrics and Gynecology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
Juan G
JG
Alvarez
Rakesh K
RK
Sharma
Mario
M
Ollero
Ramadan A
RA
Saleh
Mari C
MC
Lopez
Anthony J
AJ
Thomas
Donald P
DP
Evenson
Ashok
A
Agarwal
D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000737:chemistry; D004249:DNA Damage; D006801:Humans; D007248:Infertility, Male / Q000235:genetics*; D007962:Leukocytes / Q000737:chemistry*; D008297:Male; D009691:Nucleic Acid Denaturation; D011446:Prospective Studies; D017382:Reactive Oxygen Species; D012661:Semen / Q000166:cytology*; D013091:Spermatogenesis; D013094:Spermatozoa / Q000166:cytology*
D002843:Chromatin; D017382:Reactive Oxygen Species