ADULT UROLOGY CME ARTICLE HUMAN CYTOCHEM I zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA ARMAND 211: ELSEVIER SPERM NADH AND NADPH DIAPHORASE STRY: CORRELATION WITH SPERM MOTILITY NI, MOIRA K. O’BRYAN, LAUREN ISRAEL, AND PETER N. SCHLEGEL ABSTRACT Objectives. We have examined the correlation between the retention of residual sperm cytoplasm and sperm motility in semen from men presenting for infertility evaluation. Methods. Semen samples (n = 12) were obtained from nonazoospermic men presenting for infertility evaluation at our institution. Samples were fractionated into high-, intermediate-, and low-density subpopu- lations by Percoll gradients in order to examine the correlation between the retention of residual sperm cytoplasm and sperm motility. Residual sperm cytoplasm retention was detected by cytochemical staining of sperm for nicotinamide adenine dinucleotide (NADH)- or nicotinamide adenine dinucleotide phosphate (NADPH)-dependent diaphorase activity. Results. The different sperm subpopulations (low, intermediate, and high density) had significantly different percentages of sperm with droplet retention (analysis of variance, P ~0.05). Using either NADH or NADPH diaphorase staining as a marker of the cytoplasmic space, a significant negative correlation was observed between the percentage of sperm with residual cytoplasmic droplets and the percentage of motile sperm (r = -0.58 and -0.61, respectively, P ~0.05). Conclusions. Assessment of residual sperm cytoplasm retention is a simple diagnostic test. Although this test is of unproven value in the management of infertile men, this and other studies suggest that it may provide useful data on sperm function. UROLOGY 51: 464-468, 1998. 0 1998, Elsevier Science Inc. All rights reserved. zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA T he detrimental effects of reactive oxygen spe- cies (ROS) on spermatozoa are well recog- nized.++ Independent studies have confirmed that human spermatozoa have the capacity to generate ROS.5,6 These and other studies have also shown that excess production of ROS in semen is clearly associated with male infertility.2,5,6 The clinical rel- evance is highlighted by the finding that significant levels of ROS are detectable in the semen of up to 25% of infertile men, whereas fertile men do not produce detectable levels of ROS in their semen.2,7 Separation of cellular elements from the semen revealed that seminal plasma does not produce From the Division of Urology, University of Toronto, Toronto, Canada; Monash University Institute ofReproduction and Devel- opment, Clayton, Australia; andJames Buchanan Brady Founda- tion, Department of Urology, The New York Hospital-Cornell Medical Center and The Population Council, Centerfor Biomed- ical Research, New York, New York Reprint requests: Armand Zini, M.D., Mount Sinai Hospital, 600 University Avenue, Suite 45.5, Toronto, Ontario, Canada M5G 1X5 Submitted: August 15, 1997, accepted (with revisions): Octo- ber 1, 1997 0 1998, ELSEVIER SCIENCE INC. 464 AU RIGHTS RESERVED ROS, but in fact, spermatozoa and leukocytes (ac- tivated neutrophils) are the source of ROS.5,8 In general, it is primarily those sperm with abnormal morphology that are believed to generate high lev- els of ROS. Recent work has shown that spermato- zoa that have retained excess cytoplasm may be responsible for high semen ROS production. This excess cytoplasm contains a number of enzymes, many of which are probably detrimental to normal sperm function. ~0 It has been shown that the nic- otinamide adenine dinucleotide (NADH)-depen- dent diaphorase (or oxidase) activity, which is as- sociated with residual sperm cytoplasm, may be responsible for high semen ROS production.g-ll Gomez et al. have reported that in a subset of men with unproven fertility, the retention of residual sperm cytoplasm is positively correlated with ROS production in semen and negatively correlated with sperm motility. l l We have examined the possible influence of re- sidual sperm cytoplasm on human sperm motility in a subset of men presenting for infertility. We have assessed the retention of residual sperm cyto- plasm using cytochemical stains for NADH- and 0090-4295l98LS19.00 PI1 SOO90-4295(97)00631-6