Evaluation of poly(ADP-ribose) polymerase cleavage (cPARP) in ejaculated human sperm fractions after induction of apoptosis.
OBJECTIVE
To examine the presence of cleaved poly(ADP-ribose) polymerase(s) (cPARP) in ejaculated spermatozoa and determine cPARP levels following exposure to chemical or oxidative stress.


METHODS
Prospective pilot study.


METHODS
Tertiary care academic hospital.


METHODS
Eight healthy men.


METHODS
Semen specimens were collected, prepared with double-density-gradient centrifugation, and divided into control, hydrogen peroxide (H(2)O(2)), H(2)O(2) + 3-aminobenzamide (3-ABA), staurosporine (STS), and STS + 3-ABA treated groups.


METHODS
Cleaved PARP and apoptosis markers by flow cytometry.


RESULTS
Cleaved PARP was detected in both neat and mature fractions. The cPARP levels were similar in both mature and immature spermatozoa. In combined mature and immature fractions, a higher percentage of late apoptotic sperm was seen in STS + 3-ABA versus STS. Higher levels of late apoptotic spermatozoa were seen in immature versus mature fractions within STS and STS + 3-ABA groups. Lower levels of cPARP were seen in immature versus mature fractions in H(2)O(2) and H(2)O(2) + 3-ABA treated groups. Cleaved PARP was related to activated caspase-3.


CONCLUSIONS
Cleaved PARP is present in ejaculated human spermatozoa. Poly(ADP-ribose) polymerase inhibitors may play a different role in chemical versus oxidative stress-induced sperm damage.
Mahfouz
Reda Z
RZ
Reproductive Research Center, Glickman Urological and Kidney Institute, Department of Obstetrics and Gynecology, Cleveland Clinic, Cleveland, Ohio 44195, USA.
Sharma
Rakesh K
RK
Poenicke
Kerstin
K
Jha
Rajesh
R
Paasch
Uwe
U
Grunewald
Sonja
S
Agarwal
Ashok
A
D017209:Apoptosis / Q000502:physiology*; D001549:Benzamides / Q000494:pharmacology; D002499:Centrifugation, Density Gradient; D004249:DNA Damage / Q000187:drug effects; D004542:Ejaculation; D005434:Flow Cytometry; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006868:Hydrolysis; D020287:In Situ Nick-End Labeling; D008297:Male; D018384:Oxidative Stress; D011065:Poly(ADP-ribose) Polymerases / Q000187:drug effects / Q000378:metabolism*; D012661:Semen / Q000166:cytology / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D019311:Staurosporine / Q000494:pharmacology
D001549:Benzamides; C025160:3-aminobenzamide; D006861:Hydrogen Peroxide; D011065:Poly(ADP-ribose) Polymerases; D019311:Staurosporine