Evaluation of poly(ADP-ribose) polymerase cleavage (cPARP) in ejaculated human sperm fractions after induction of apoptosis
Received 30 January 2008; received in revised form 28 February 2008; accepted 28 February 2008. published online 23 May 2008.
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.
Design
Prospective pilot study.
Setting
Tertiary care academic hospital.
Patient(s)
Eight healthy men.
Intervention(s)
Semen specimens were collected, prepared with double-density-gradient centrifugation, and divided into control, hydrogen peroxide (H2O2), H2O2 + 3-aminobenzamide (3-ABA), staurosporine (STS), and STS + 3-ABA treated groups.
Main Outcome Measure(s)
Cleaved PARP and apoptosis markers by flow cytometry.
Result(s)
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 H2O2 and H2O2 + 3-ABA treated groups. Cleaved PARP was related to activated caspase-3.
Conclusion(s)
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.
aReproductive Research Center, Glickman Urological and Kidney Institute, and Department of Obstetrics and Gynecology, Cleveland Clinic, Cleveland, Ohio
bDepartment of Andrology, University of Leipzig, Leipzig, Germany
Reprint requests: Ashok Agarwal, Ph.D, H.C.L.D., Professor and Director, Reproductive Research Center, Staff, Glickman Urological and Kidney Institute, Department of Obstetrics and Gynecology, Cleveland Clinic, 9500 Euclid Avenue, Desk A19.1, Cleveland, Ohio 44195 (FAX: 216-445-6049).
R.M. has nothing to disclose. R.S. has nothing to disclose. P.K. has nothing to disclose. R.J. has nothing to disclose. U.P. has nothing to disclose. S.G. has nothing to disclose. A.A. has nothing to disclose.
Presented at the 63rd Annual Meeting of the American Society of Reproductive Medicine, Washington, DC, October 13–19, 2007.
Supported by a research grant from the Research Programs Committee, Cleveland Clinic (RPC no. 1067, IRB 8530).
ABSTRACT