Evaluation of poly(ADP-ribose) polymerase cleavage (cPARP) in ejaculated human sperm fractions after induction of apoptosis Reda Z. Mahfouz, M.D., Rakesh.

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Presentation transcript:

Evaluation of poly(ADP-ribose) polymerase cleavage (cPARP) in ejaculated human sperm fractions after induction of apoptosis Reda Z. Mahfouz, M.D., Rakesh K. Sharma, Ph.D., Kerstin Poenicke, Rajesh Jha, Ph.D., Uwe Paasch, M.D., Sonja Grunewald, M.D., Ashok Agarwal, Ph.D., H.C.L.D. Fertility and Sterility Volume 91, Issue 5, Pages 2210-2220 (May 2009) DOI: 10.1016/j.fertnstert.2008.02.173 Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Various steps involved in processing of semen sample after collection, modulation and analysis for various markers by flow cytometry. 3-ABA = 3-aminobenzamide; cPARP = cleaved poly(ADP-ribose) polymerase; FACS = fuorescence-activated cell sorter; PARP = poly(ADP-ribose) polymerase; PI = propidium iodide; TUNEL = terminal deoxynucleotidyl transferase–mediated fluorescein-dUTP nick-end labeling. Fertility and Sterility 2009 91, 2210-2220DOI: (10.1016/j.fertnstert.2008.02.173) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Cleaved poly(ADP-ribose) polymerase (cPARP)+ve sperm overlay histograms: (A) Mature vs. immature with the negative control; (B) oxidative vs. chemical stress–induced cell damage with unstained negative control in mature spermatozoa; (C) oxidative vs. chemical stress–induced cell damage with unstained negative control in immature spermatozoa; (D) effect of PARP inhibition in oxidative or chemical stress–induced cell damage on PARP cleavage. Fertility and Sterility 2009 91, 2210-2220DOI: (10.1016/j.fertnstert.2008.02.173) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 3 TUNEL+ve in mature (A) and immature (B) sperm fractions after chemical and oxidative stress–induced damage in presence or absence of PARP inhibitor; cPARP+ve in mature (C) and immature (D) sperm fractions after chemical and oxidative stress–induced damage in presence or absence of PARP inhibitor. STS = staurosporine; other abbreviations as in Figures 1 and 2. Fertility and Sterility 2009 91, 2210-2220DOI: (10.1016/j.fertnstert.2008.02.173) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Early apoptosis in mature (A) and immature (B) sperm fraction and late apoptosis in mature (C) and immature (D) sperm fraction after chemical and oxidative stress-induced damage in presence or absence of PARP inhibitor. Fertility and Sterility 2009 91, 2210-2220DOI: (10.1016/j.fertnstert.2008.02.173) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions