Article
Double probing of human spermatozoa for persistent histones, surplus cytoplasm, apoptosis and DNA fragmentation

https://doi.org/10.1016/S1472-6483(10)60464-6Get rights and content

Abstract

Individual spermatozoa were assessed with pairs of probes for persistent histones and cytoplasmic retention, persistent histones and DNA fragmentation, and persistent histones and apoptotic markers. The individual spermatozoa were treated sequentially with combinations of probes for these cytoplasmic and nuclear biochemical markers. Sperm fields were recorded with computer-assisted imaging, and staining patterns with the two probes in the same spermatozoa were examined and scored as light, intermediate or dark (mature to arrested-maturity spermatozoa). The effects of arrested sperm maturation were similar with respect to the cytoplasmic and nuclear characteristics of spermatozoa in 84% of cells, indicating that cytoplasmic and nuclear attributes of arrested sperm maturation are related. However, there were moderate (intermediate–dark or intermediate–light patterns, 14.5% of cells) or major (light–dark patterns, 1.6% of cells) discrepancies in the intensity of the double staining patterns. Thus, testing with single maturity markers may not be fully reliable. These findings are important with respect to: (i) arrested sperm maturation; (ii) potential efficacy of antioxidant and similar therapeutic strategies in subfertile men, as spermatozoa with infrastructure defects due to mismaturation or maturation arrest are unlikely to respond to interventions; and (iii) detection of adverse male environmental exposures.

Section snippets

Leyla Sati MSc is currently a PhD student in the Department of Histology and Embryology, Akdeniz University, Turkey, having previously worked as a pre-doctoral Fellow in the Sperm Physiology Laboratory, Yale University School of Medicine with Dr Gabor Huszar. Her research interests include the various biochemical markers of human sperm maturity and function, and the relationship between tyrosine phosphorylation in human spermatozoa and sperm binding to the zona pellucida and hyaluronic acid.

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    Leyla Sati MSc is currently a PhD student in the Department of Histology and Embryology, Akdeniz University, Turkey, having previously worked as a pre-doctoral Fellow in the Sperm Physiology Laboratory, Yale University School of Medicine with Dr Gabor Huszar. Her research interests include the various biochemical markers of human sperm maturity and function, and the relationship between tyrosine phosphorylation in human spermatozoa and sperm binding to the zona pellucida and hyaluronic acid. She has been nominated for and awarded prizes at international meetings, including the American Society of Reproductive Medicine and the 2007 International Dermatology and Andrology Conference in Egypt.

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