Characterization of human sperm populations using conventional parameters, surface ubiquitination, and apoptotic markers

Abstract

Objective: To directly compare distinct assays proposed to monitor human sperm quality and possibly preselect sperm populations for assisted reproductive technology (ART). Design: Analysis of human sperm sample quality using several methodologies. Setting: Academic and clinical institutions. Patient(s): Samples from consenting patients undergoing routine semen analysis or ART. Interventions: Human sperm samples were analyzed in terms of World Health Organization parameters and processed for annexin V, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling of DNA (TUNEL), and the sperm-ubiquitin tag immunoassay (SUTI). Samples were analyzed both by flow cytometry and fluorescence microscopy. Main Outcome Measure(s): Correlations among apoptotic markers (outer leaflet phosphatidylserine exposure, membrane integrity, and DNA fragmentation), external ubiquitination, and semen parameters in human spermatozoa. Result(s):Nonviable sperm, TUNEL-positive cells, and ubiquitin fluorescence intensity means inversely correlate with semen parameters. Apoptotic markers do not correlate with sperm surface ubiquitination. Normozoospermic samples have a higher number of viable cells and lower DNA fragmentation compared with samples with abnormal parameters. Nonviable sperm are more prevalent in samples with low counts and poor morphology but not low motility. Not all sperm with morphologic abnormalities present surface ubiquitination. Conclusion(s):Sperm quality is inversely correlated with lack of viability, DNA fragmentation, and ubiquitin fluorescence intensity means. However, none of the apoptotic markers correlate with ubiquitin labeling. Elimination of defective sperm cells prior to ART using surface markers (annexin V, ubiquitin) seems unwarranted at this stage. (Fertil Steril 2007;87:572-83.

Evaluation of ubiquitin and annexin V in sperm population selected based on density gradient centrifugation and zeta potential (DGC-Zeta)

“…Therefore, our results are also consistent with this report which shows a concomitant increase in ubiquitination and annexin staining [36].…”

Mitochondria functionality and sperm quality

“…PS exposure can be monitored using fluorescent Annexin V in unpermeabilised (live) cells. In fact, Annexin V staining revealed more viable cells in normozoospermic patients (Varum et al 2007) and seemed to correlate with sperm parameters in other studies (Shen et al 2002, Weng et al 2002. Importantly, the use of magnetic activated cell sorting (MACS) with Annexin V microbeads to select sperm reduced the percentage of altered cells (Lee et al 2010, Rawe et al 2010, Tavalaee et al 2012.…”

Critical Aspects of Detection of Sperm DNA Fragmentation by Tunel/Flow Cytometry

“…By reviewing studies conducted in raw semen from similar groups Muciaccia et al 2007;O'Flaherty et al 2008;Cohen-Bacrie et al 2009;Tunc et al 2008;Martin et al 2005Formaldeyde d Muratori et al 2003Caglar et al 2007;Tarozzi et al 2009;Domínguez-Fandos et al 2007;Young et al 2003;Avendañ o et al 2009a and2009b;Stronati et al 2006;Ramos et al 2008;Tesarik et al 2004 Permeabilization with detergents Absent Martin et al 2005;O'Flaherty et al 2008;Young et al 2003;Said et al 2006Present e Chohan et al 2006Aoki et al 2006;Caglar et al 2007;Varum et al 2007;Muratori et al 2008a andTunc et al 2008;Frydman et al 2008;Avendañ o et al 2009a and2009b Labelling Direct labeling f O'Flaherty et al 2008;Zhang et al 2008;Aoki et al 2006;Torregrosa et al 2006;Young et al 2003;Muratori et al 2008a;.Greco et al 2005;Paasch et al 2004;Brugnon et al 2006;Donnelly et al 2000 Indirect labeling g Varum et al 2007;Sergerie et al 2005;P...…”