Relationship between caspase activity and apoptotic markers in human sperm in response to hydrogen peroxide and progesterone

• Published in: Journal of Reproduction and Development 2009, Vol.55(6), pp.615-621
• Main Authors: Lozano, G.M., University of Extremadura, Badajoz (Spain). Infantile Hospital, Bejarano, I, Espino, J, Gonzalez, D, Ortiz, A, Garcia, J.F, Rodriguez, A.B, Pariente, J.A
• Format: Article
• Language: English
• Subjects: Adult; Annexin A5 - metabolism; APOPTOSE; APOPTOSIS; Apoptosis - drug effects; Biomarkers; CALCIO; CALCIUM; Calcium Signaling - drug effects; Caspases; Caspases - metabolism; Cell Membrane - metabolism; DNA Fragmentation - drug effects; ENZIMAS; ENZYME; ENZYMES; ESPERMATOGENESIS; ESPERMATOZOO; http://www.fao.org/aos/agrovoc#c_1196; http://www.fao.org/aos/agrovoc#c_2603; http://www.fao.org/aos/agrovoc#c_36634; http://www.fao.org/aos/agrovoc#c_3725; http://www.fao.org/aos/agrovoc#c_6218; http://www.fao.org/aos/agrovoc#c_7285; http://www.fao.org/aos/agrovoc#c_7287; Humans; HYDROGEN PEROXIDE; Hydrogen Peroxide - pharmacology; In Situ Nick-End Labeling; Infertility, Male - physiopathology; Male; Middle Aged; PEROXIDO DE HIDROGENO; PEROXYDE D'HYDROGENE; Phosphatidylserine exposure; Phosphatidylserines - metabolism; PROGESTERONA; PROGESTERONE; Progesterone - pharmacology; SPERMATOGENESE; SPERMATOGENESIS; SPERMATOZOA; Spermatozoa - drug effects; SPERMATOZOIDE; TUNEL; Young Adult
• ISSN: 0916-8818; 1348-4400
• DOI: 10.1262/jrd.20250

Apoptosis plays an essential role in normal spermatogenesis, but deregulations of this biological process, which is closely associated with male infertility, have been found. Whereas calcium homeostasis is a key regulator of cell survival, sustained elevation of intracellular calcium plays a role in apoptosis. The aim of this research was to determine the role of two different calcium mobilizing agents, hydrogen peroxide (H2O2) and the physiological agonist progesterone, on the apoptosis process of human ejaculated spermatozoa. Translocation of membrane phosphatidylserine was examined with an annexin V binding assay, DNA damage was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL assay) and caspase-3 activity was assessed using a fluorometric assay. After incubation of spermatozoa for 1 h with either 10 micro M H2O2 or 20 microM of progesterone, there was a significant increase in both caspase-3 activity and the percentage of annexin V-positive cells. Similarly, the TUNEL results were significantly higher 1 h after incubation with either 10 micro M H2O2 or 20 micro M of progesterone. In fact, progesterone-treated cells showed a three-fold increase (from 17.6 to 52.9%) of TUNEL-positive cells compared to untreated cells, while H2O2-treated cells exhibited a two-fold increase (from 17.6 to 37.9%). In sum, our results suggest that spermatozoa treated with calcium mobilizing agents, such as H2O2 and progesterone, seem to undergo an apoptosis process that is dependent on caspase-3 activation.