Erica Miraglia, Ph. D. , Cristina Lussiana, B. Sc

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The pentose phosphate pathway plays an essential role in supporting human sperm capacitation Erica Miraglia, Ph.D., Cristina Lussiana, B.Sc., Daniele Viarisio, Ph.D., Cinzia Racca, B.Sc., Alessia Cipriani, B.Sc., Elena Gazzano, Ph.D., Amalia Bosia, M.D., Alberto Revelli, M.D., Dario Ghigo, M.D. Fertility and Sterility Volume 93, Issue 7, Pages 2437-2440 (May 2010) DOI: 10.1016/j.fertnstert.2009.09.005 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) The expression of glucose-6-phosphate dehydrogenase (G6PD) mRNA was investigated by reverse-transcription polymerase chain reaction in freshly ejaculated samples after 2 hours of capacitating conditions (CAP) or noncapacitating conditions (NC), in the absence or presence of menadione (MEN, 100 μmol/L) or of the protein synthesis inhibitor cycloheximide (chx, 20 μg/mL). The figure is representative of three experiments with similar results. β-Actin mRNA was checked as product of a housekeeping gene. (B) Effect of apocynin, diphenylene iodonium (DPI), and DHEA on pentose phosphate pathway (PPP) activity in NC and CAP human spermatozoa, in the absence (CTRL) or presence of 300 μmol/L apocynin (APO), 10 μmol/L DPI, or 500 μmol/L DHEA. The PPP activity is expressed as percentage of control PPP activity. Each measurement was performed in duplicate, and data are presented as mean ± SEM (n = 5). ∗P<.0001 vs. NC in the corresponding experimental condition. °P<.0001 vs. CAP CTRL. (C) Effect of apocynin, DPI, and DHEA on the CTC staining patterns of human spermatozoa. Human fresh samples after 2 h of capacitating conditions (+) or noncapacitating conditions (−), were treated with 300 μmol/L apocynin (APO), 10 μmol/L DPI or 500 μmol/L DHEA for 30 minutes and used for the chlortetracycline staining assay. Each experiment was performed in quadruplicate, and values are presented as mean ± SEM (n = 7). ∗P<.001 vs. CTRL in the absence of the capacitating medium (in the corresponding staining pattern); °P<.001 vs. CTRL in the presence of the capacitating medium (in the corresponding staining pattern). Fertility and Sterility 2010 93, 2437-2440DOI: (10.1016/j.fertnstert.2009.09.005) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions