不同浓度Ca2+对塔里木马鹿冻融精子体外获能及蛋白酪氨酸磷酸化的影响

doi:10.16431/j.cnki.1671-7236.2016.08.027

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (8): 2128-2134.doi: 10.16431/j.cnki.1671-7236.2016.08.027

不同浓度Ca²⁺对塔里木马鹿冻融精子体外获能及蛋白酪氨酸磷酸化的影响

木沙·托合提¹, 张俊杰¹, 艾海提¹, 李和平², 华兴耀³, 敬斌宇⁴, 库尔班·吐拉克¹

1. 新疆农业大学动物科学学院, 乌鲁木齐 830052;
2. 东北林业大学野生动物资源学院, 哈尔滨 150040;
3. 新疆昌吉市盛华马鹿驯养繁育基地, 昌吉 831100;
4. 新疆厚拾生物科技有限责任公司, 库尔勒 841001

收稿日期:2015-12-17 出版日期:2016-08-20 发布日期:2016-08-23
通讯作者: 库尔班·吐拉克 E-mail:kturak@sina.com
作者简介:木沙·托合提(1991-),男,新疆人,硕士生,研究方向:动物遗传育种与繁殖,E-mail:1350896482@qq.com
基金资助:
新疆维吾尔自治区自然科学基金项目（2013211A029）

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. 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

摘要/Abstract

摘要：

为探讨不同浓度Ca²⁺对马鹿精子体外获能的影响，本研究以塔里木马鹿冻融精子为试验材料，将精子分别悬浮于含不同浓度Ca²⁺（0、1.1、2.2、3.5、5.0 mmol/L）的台氏液（sp-TALP液）中，在培养0、2、4 h时，采用金霉素（CTC）染色法评价精子获能状态，采用SDS-PAGE分离精子膜蛋白，进行免疫印迹分析，检测酪氨酸磷酸化蛋白的表达水平。结果表明，Ca²⁺浓度为1.1、2.2 mmol/L有利于精子活力的维持（P<0.05），精子获能率极显著高于对照组和高浓度组（3.5、5.0 mmol/L；P<0.01），精子存活时间最长（P<0.01），但高浓度Ca²⁺（5.0 mmol/L）对精子活力具有显著抑制作用（P<0.05），精子获能率极显著低于低浓度组（P<0.01），精子存活时间最短（P<0.01）；另外，随着培养时间的推移精子发生酪氨酸磷酸化蛋白的表达水平有所不同，培养2、4 h时，1.1 mmol/L组精子蛋白磷酸化水平极显著高于其他各组（P<0.01），高浓度Ca²⁺（3.5、5.0 mmol/L）组酪氨酸磷酸化蛋白的表达水平极显著下降（P<0.01）。结果表明，塔里木马鹿精子体外获能所需的适宜Ca²⁺浓度为1.1 mmol/L，且获能过程中Ca²⁺的存在是必要的。

关键词: 塔里木马鹿; 精子获能; Ca²⁺; 蛋白酪氨酸磷酸化

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

中图分类号:

S814

木沙·托合提, 张俊杰, 艾海提, 李和平, 华兴耀, 敬斌宇, 库尔班·吐拉克. 不同浓度Ca²⁺对塔里木马鹿冻融精子体外获能及蛋白酪氨酸磷酸化的影响[J]. 《中国畜牧兽医》, 2016, 43(8): 2128-2134.

MUSHA Tuoheti, ZHANG Jun-jie, AI Hai-ti, LI He-ping, HUA Xing-yao, JING Bin-yu, KUERBAN Tulake. Effect of Different Concentrations of Ca²⁺ on in vitro Capacitation and Protein Tyrosine Phosphorylation of Tarim Red Deer (Cervus elaphus yarkandensis) Frozen-thawed Sperm[J]. , 2016, 43(8): 2128-2134.

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不同浓度Ca²⁺对塔里木马鹿冻融精子体外获能及蛋白酪氨酸磷酸化的影响

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

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