›› 2016, Vol. 43 ›› Issue (8): 2128-2134.doi: 10.16431/j.cnki.1671-7236.2016.08.027

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Effect of Different Concentrations of Ca2+ on in vitro Capacitation and Protein Tyrosine Phosphorylation of Tarim Red Deer (Cervus elaphus yarkandensis) Frozen-thawed Sperm

MUSHA Tuoheti1, ZHANG Jun-jie1, AI Hai-ti1, LI He-ping2, HUA Xing-yao3, JING Bin-yu4, KUERBAN Tulake1   

  1. 1. College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China;
    2. College of Wildlife Resources, Northeast Forestry University, Harbin 150040, China;
    3. Xinjiang Changji Shenghua Domesticated Wapiti Breeding Base, Changji 831100, China;
    4. Xinjiang Houshi Biotechnology Co., Ltd., Korla 841001, China
  • Received:2015-12-17 Online:2016-08-20 Published:2016-08-23

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Abstract:

In order to investigate the effect of different concentrations of Ca2+ on in vitro capacitation of wapiti sperm,frozen-thawed sperm of Tarim Red deer (Cervus elaphus yarkandensis) was used as experimental materials in this study.Sperm capacitation status were assessed by chlortetracycline (CTC) staining,the expression levels of tyrosine phosphorylated protein were detected with Western blotting analysis of sperm membrane protein separated by SDS-PAGE,following the sperm were suspended in sp-TALP liquids,which contained different concentrations of Ca2+(0,1.1,2.2,3.5 and 5.0 mmol/L),and cultured for 0,2,4 h.The results showed that the low concentration of Ca2+ (1.1,2.2 mmol/L) was conducive to the maintenance of sperm motility (P<0.05),sperm capacitation rate was extremely significant higher than that of control group and high concentration groups (3.5,5.0 mmol/L;P<0.01),sperm survival time was the longest (P<0.01),but high concentration of Ca2+ (5.0 mmol/L) significantly inhibited sperm motility (P<0.05),sperm capacitation rate was extremely significantly lower than that in low concentration group (P<0.01),sperm survival time was the shortest (P<0.01). In addition,the expression levels of tyrosine phosphorylated proteins were different,and the levels of sperm protein phosphorylation in low concentration group (1.1 mmol/L) were extremely significant higher than those in other groups (P<0.01),and the expression levels of tyrosine phosphorylated proteins in high concentration groups of Ca2+ (3.5,5.0 mmol/L) were extremely significantly decreased (P<0.01). These results suggested that the appropriate Ca2+ concentration required for Tarim Red deer sperm in vitro capacitation was 1.1 mmol/L,and the presence of Ca2+ in process of capacitation was necessary.

Key words: Cervus elaphus yarkandensis; sperm capacitation; Ca2+; protein tyrosine phosphorylation

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