Evaluation of HSPA2 in fertile and infertile individuals

Abstract

Heat-shock protein A2 (HSPA2) is a testis-specific member of the HSP70 family known to correlate with sperm maturity, function and fertility. The aim of this study was to compare expression of HSPA2 in fertile and infertile individuals using a recently marketed highly purified polyclonal antibody. Thus, after analysing sperm concentration, motility and morphology in semen sample of 49 individuals with male factor infertility and 47 fertile individuals according to World Health Organization guidelines, we evaluated HSPA2 by microscope florescence, flow cytometry, Western blot and RT-PCR. We observed higher percentage of sperm expressing HSPA2 in anterior and equatorial regions in fertile than infertile individuals by microscope fluorescence. Percentage of sperm expressing HSPA2 in two conditions (fixed permeabilised and capacitated) by flow cytometry showed that percentage of HSPA2-positive sperm in fixed permeabilised and also capacitated samples was significantly different between fertile and infertile individuals. Western blot analysis of HSPA2 in semen samples revealed high variation within the fertile and infertile population. The results of RT-PCR revealed higher expression of HSPA2 in the fertile compared to infertile individuals, but this difference was not significant. According to the results, we suggest that HSPA2 expression is heterogeneously expressed on different part of fixed permeabilised sperm and its expression is significantly higher in fertile compared to infertile individuals. The surface expression of this protein significantly increases following capacitation in both fertile and infertile individuals. HSPA2 expression significantly correlates with sperm concentration and morphology. Therefore, aberrant HSPA2 expression may play an important role in capacitation and fertilisation processes.

Aquaporin-11 control of testicular fertility markers in Syrian hamsters

“…Expression of human CATSPER1 and murine Catsper1 are required for sperm hyperactivation and fertility (Lishko et al, 2011; Carlson et al, 2003). Low expression levels of heat shock protein A2 ( HSPA2 ) in human semen have been associated with infertility (Motiei et al, 2012) and previous studies suggest murine Hspa2 is required for meiosis and spermatogenesis (Zhu et al, 1997). Phosphoglycerate kinase 2 ( Pgk2 ), which catalyzes an ATP-generating step in glycolysis and is expressed only during spermatogenesis, has been found to be essential for normal sperm motility and male fertility in mice (Danshina et al, 2010).…”

“…In addition to that of Aqp11 , expression levels of Hspa2 , Sept12 , and Tnp2 were reduced by Aqp11 siRNA. Importantly, roles for Hspa2 , Sept12 , and Tnp2 in promoting sperm motility and spermatogenesis is conserved in humans and rodents (Zhao et al, 2001; Adham et al, 2001; Motiei et al, 2012; Kuo et al, 2012; Lin et al, 2009; Dix et al, 1997). Interestingly, this same treatment increased expression of steroidogenic 3 β Hsd .…”

Expression profile ofPLCζ,PAWP,andTR-KITin association with fertilization potential, embryo development, and pregnancy outcomes in globozoospermic candidates for intra-cytoplasmic sperm injection and artificial oocyte activation

“…Membranes were hybridized with primary antibodies (GAPDH and PAWP for 90 min at room temperature; PLCf and TR-KIT for overnight at 4°C). After washing thrice, bands were detected following 1 h incubation with secondary antibodies (goat anti-rabbit IgG-HRP and peroxidase-conjugated goat antimouse immunoglobulins) (Motiei et al, 2013).…”

Toward the identification of a subset of unexplained infertility: a sperm proteomic approach

“…They are members of the HSP70 proteins, and both have been identified on the surface of human sperm (29). HSPA2 is involved in sperm maturity and function as well as the regulation of sperm-egg recognition (30)(31)(32). HSPA5 is the major HSP70 protein of the endoplasmic reticulum and has a crucial role in the assembly of endoplasmic reticulum proteins and protein complexes as well as the unfolded protein response (33).…”