A proteomic analysis on human sperm tail: comparison between normozoospermia and asthenozoospermia

Abstract

Purpose Asthenozoospermia is a common cause of human male infertility characterized by reduced sperm motility. The molecular mechanism that impairs sperm motility is not fully understood. This study proposed to identify novel biomarkers by focusing on sperm tail proteomic analysis of asthenozoospermic patients. Methods Sperm were isolated from normozoospermic and asthenozoospermic semen samples. Tail fractions were obtained by sonication followed by Percoll gradient. The proteins were extracted by solubilization and subjected to two-dimensional gel electrophoresis (2-DE); then, the spots were analyzed using Image Master 2D Platinum software. The significantly increased/ decreased amounts of proteins in the two groups were exploited by matrix-assisted laser desorption-ionization time-of-flight/ time-of-flight (MALDI-TOF-TOF) mass spectrometry. Results Three hundred ninety protein spots were detected in both groups. Twenty-one protein spots that had significantly altered amounts (p<0.05) were excised and exploited using MALDI-TOF-TOF mass spectrometry. They led to the identification of the following 14 unique proteins: Tubulin beta 2B; glutathione S-transferase Mu 3; keratin, type II cytoskeletal 1; outer dense fiber protein 2; voltage-dependent anionselective channel protein 2; A-kinase anchor protein 4; cytochrome c oxidase subunit 6B; sperm protein associated with the nucleus on the X chromosome B; phospholipid hydroperoxide glutathione peroxidase-mitochondrial; isoaspartyl peptidase/L-asparaginase; heat shock-related 70 kDa protein 2; stress-70 protein, mitochondrial; glyceraldehyde-3-phosphate dehydrogenase, testis-specific and clusterin. Conclusion Fourteen proteins present in different amounts in asthenozoospermic sperm tail samples were identified, four of which are reported here for the first time. These proteins might be used as markers for the better diagnosis of sperm dysfunctions, targets for male contraceptive development, and to predict embryo quality.

Could the glycosylation analysis of seminal plasma clusterin become a novel male infertility biomarker?

“…Zalata et al (2012) showed that it correlated negatively with, inter alia, sperm count, motility, linear velocity, and significantly correlated positively with the percentage of abnormal forms of spermatozoa and sperm DNA fragmentation. Hashemitabar, Sabbagh, Orazizadeh, Ghadiri, and Bahmanzadeh (2015) compared the concentration of sperm tail proteins in normozoospermic and astenozoospermic men. The experiment showed increased levels of 14 proteins in astenozoospermic patients, CLU among them.…”

Towards the identification of reliable sperm biomarkers for male infertility: A sperm proteomic approach

“…It has been hypothesised that the sperm protein profile in infertile men is altered, and some researchers have assessed these profiles in several male infertility conditions including varicocele (Agarwal, Sharma, Samanta, Durairajanayagam, & Sabanegh, ; Agarwal, Sharma, Durairajanayagam, Ayaz, et al., ; Agarwal, Sharma, Durairajanayagam, et al., ; Hosseinifar et al., ), obesity (Kriegel et al., ; Liu et al., ), smoking (Chen et al., ), failed fertilisation after assisted reproduction techniques (ART; Frapsauce et al., ; Legare et al., ; Pixton et al., ), asthenozoospermia (Amaral et al., ; Hashemitabar, Sabbagh, Orazizadeh, Ghadiri, & Bahmanzadeh, ; Liu et al., ; Martinez‐Heredia, de Mateo, Vidal‐Taboada, Ballesca, & Oliva, ; Parte et al., ; Shen, Wang, Liang, & He, ; Siva et al., ; Zhao et al., ) and idiopathic infertility (Legare et al., ; McReynolds et al., ; Xu et al., ). Sperm protein profiles have also been evaluated in men with semen oxidative stress (Ayaz et al., ; Sharma et al., ) and sperm DNA fragmentation (Intasqui et al., ; de Mateo et al., ), which are common causes of male infertility.…”

Differential gene regulation in selected lines for high and low sperm production in male guppies

“…LOC103474644 (product ID: XP_008423986) on chromosome 2, showed higher expression ( p adj = .0008, LogFC = 0.95) in the LS than the HS line and it codifies for a keratin Type II cytoskeletal protein (KRT) which has been found to be upregulated in asthenozoospermic human males (Hashemitabar, Sabbagh, Orazizadeh, Ghadiri, & Bahmanzadeh, 2015).…”

“…Only three genes were significantly upregulated in the LS line but in all three cases, the fold change between selection lines was low. The most interesting of these genes was KRT, which has higher expression levels in asthenozoospermic human male patients than in normal sperm donors (Hashemitabar et al, 2015). In humans, it seems that the higher expression of KRT in asthenozoospermia is a compensatory effect on lower levels of tyrosine phosphorylation (Hashemitabar et al, 2015).…”

“…The most interesting of these genes was KRT, which has higher expression levels in asthenozoospermic human male patients than in normal sperm donors (Hashemitabar et al, 2015). In humans, it seems that the higher expression of KRT in asthenozoospermia is a compensatory effect on lower levels of tyrosine phosphorylation (Hashemitabar et al, 2015). A compensatory effect of the LS production seems to characterize the upregulation of the gene sets identified by the GSEA analysis also.…”