The effect of anti-eppin antibodies on ionophore A23187-induced calcium influx and acrosome reaction of human spermatozoa

Zhang, Jie; Ding, Xinliang; Bian, Zenghui; Xia, Yankai; Lu, Chuncheng; Wang, Shoulin; Lee, Song; Wang, Xinru

doi:10.1093/humrep/dep356

Cited by 23 publications

(18 citation statements)

References 32 publications

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“…EPPIN, characterized by both WAP-type and Kunitz-type consensus sequences (Wang et al 2005, O'Rand et al 2007, acts a decapacitating factor by modulating the activity of PSA in semenogelin, which inhibits human sperm capacitation (de Lamirande et al 2001). However, it has been found that anti-EPPIN antibodies significantly inhibited the human sperm AR induced by A23187, reduced intracellular Ca 2C concentration, and did not change tyrosine phosphorylation of sperm proteins (Zhang et al 2010). Among all these molecules described as decapacitating factors and also serine protease inhibitors, it is clear that EPPIN and SERPINE2 have a role outside the sperm.…”

Section: Spink3 Involves No Signalling In Spermmentioning

confidence: 99%

SPINK3 modulates mouse sperm physiology through the reduction of nitric oxide level independently of its trypsin inhibitory activity

Zalazar

Lancellotti²,

Clementi

et al. 2012

Reproduction

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Serine protease inhibitor Kazal-type (SPINK3)/P12/PSTI-II is a small secretory protein from mouse seminal vesicle which contains a KAZAL domain and shows calcium (Ca 2C )-transport inhibitory (caltrin) activity. This molecule was obtained as a recombinant protein and its effect on capacitated sperm cells was examined. SPINK3 inhibited trypsin activity in vitro while the fusion protein GST-SPINK3 had no effect on this enzyme activity. The inactive GST-SPINK3 significantly reduced the percentage of spermatozoa positively stained for nitric oxide (NO) with the specific probe DAF-FM DA and NO concentration measured by Griess method in capacitated mouse sperm; the same effect was observed when sperm were capacitated under low Ca 2C concentration, using either intracellular (BAPTA-AM) or extracellular Ca 2C (EDTA) chelators. The percentage of sperm showing spontaneous and progesterone-induced acrosomal reaction was significantly lower in the presence of GST-SPINK3 compared to untreated capacitated spermatozoa. Interestingly, this decrease was overcome by the exogenous addition of the NO donors, sodium nitroprusside (SNP), and S-nitrosoglutathione (GSNO). Phosphorylation of sperm proteins in tyrosine residues was partially affected by GST-SPINK3, however, only GSNO was able to reverse this effect. Sperm progressive motility was not significantly diminished by GST-SPINK3 or BAPTA-AM but enhanced by the addition of SNP. This is the first report that demonstrates that SPINK3 modulates sperm physiology through a downstream reduction of endogenous NO concentration and independently of SPINK3 trypsin inhibitory activity.

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Section: Spink3 Involves No Signalling In Spermmentioning

confidence: 99%

SPINK3 modulates mouse sperm physiology through the reduction of nitric oxide level independently of its trypsin inhibitory activity

Zalazar

Lancellotti²,

Clementi

et al. 2012

Reproduction

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“…The method was performed as described previously [Zhang et al 2010]. Briefly, a suspension of the sperm (1 Â 10 6 cells) was washed twice in PBS and centrifuged (500 g, 5 min).…”

Section: Sperm Protein Extractsmentioning

confidence: 99%

“…Membranes were washed tree times in TBS and then probed with peroxidase conjugated goat anti-mouse IgG (1:1000; Beijing ZhongShan Biotechnology Co., China) for 1 h at 37 C. After washing, the specific signals were detected by a Molecular Imager ChemiDoc XRS+ (Bio-Rad, USA). The relative expression of ACO2 was calculated as the ratio of their average pixel intensity to that of the b-tubulin loading control, which was similarly described previously [Zhang et al 2010]. …”

Section: Western Blotmentioning

confidence: 99%

The role of mitochondrial aconitate (ACO2) in human sperm motility

Tang

Liu

Wang

et al. 2014

Systems Biology in Reproductive Medicine

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Deficiencies in tricarboxylic acid (TCA) cycle enzymes have been shown to cause a wide spectrum of human diseases, including malignancies and neurological and cardiac diseases. In mammalian spermatozoa mitochondria, the TCA cycle is known to be a crucial metabolic pathway that contributes to produce ATP. There is little known about the role and mechanism of mitochondrial aconitase (ACO2), which is an important regulatory enzyme of the TCA cycle, in asthenozoospermia. In the current study, immunofluorescence staining localized ACO2 to the human sperm mid-piece. By immunoblotting, we demonstrated that the level of ACO2 protein in asthenozoospermic samples was significantly decreased compared with that in normal fertile men. Importantly, we first observed that co-incubation of isocitrate with low motile sperm suspensions significantly improved sperm motility, which might be due to elevated intracellular ATP. The improvement of the sperm motility by isocitrate may have important clinical implications in the treatment of asthenozoospermia and certainly warrants further investigation.

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“…2 Functional studies of eppin have focused on the form that lacks a signal sequence. [4][5][6][7] The importance of eppin in male reproduction was confirmed by the demonstration of a reversible contraceptive effect in male monkeys immunized with recombinant human eppin and by the upregulation of EPPIN gene expression in non-obstructive azoospermia patients. 1,8 In further studies, we found that variants of the EPPIN gene affect the risk of idiopathic male infertility in the Han-Chinese population.…”

Section: Introductionmentioning

confidence: 93%

Preparation and immunogenicity of tag-free recombinant human eppin

Zhang¹,

Ding²,

Bian³

et al. 2011

Asian J Androl

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The effect of anti-eppin antibodies on ionophore A23187-induced calcium influx and acrosome reaction of human spermatozoa

Abstract: These results demonstrate that eppin can modulate intracellular calcium concentrations and subsequently affect the calcium ionophore A23187-induced acrosome reaction.

Cited by 23 publications

References 32 publications

SPINK3 modulates mouse sperm physiology through the reduction of nitric oxide level independently of its trypsin inhibitory activity

SPINK3 modulates mouse sperm physiology through the reduction of nitric oxide level independently of its trypsin inhibitory activity

The role of mitochondrial aconitate (ACO2) in human sperm motility

Preparation and immunogenicity of tag-free recombinant human eppin

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