《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (5): 1421-1428.doi: 10.16431/j.cnki.1671-7236.2019.05.020

• 遗传繁育 • 上一篇    下一篇

PP1γ2对树鼩精子成熟和运动性的调控作用研究

朱方方, 李雪, 杨明华, 吴文静, 匡海鸥, 李亚辉   

  1. 云南农业大学动物科学技术学院, 昆明 650201
  • 收稿日期:2018-09-29 出版日期:2019-05-20 发布日期:2019-05-20
  • 通讯作者: 李亚辉 E-mail:kmliyh@163.com
  • 作者简介:朱方方(1990-),女,河南许昌人,硕士生,研究方向:动物生殖生物学,E-mail:807213784@qq.com;李雪(1993-),女,云南昆明人,硕士生,研究方向:动物生殖生物学,E-mail:1416506791@qq.com
  • 基金资助:

    国家自然科学基金(31460308、31860330);云南中科灵长类生物医学重点实验室开放基金(云中科字2016-008号)

Study on the Regulation of PP1γ2 on Sperm Maturation and Motility in Tree Shrews

ZHU Fangfang, LI Xue, YANG Minghua, WU Wenjing, KUANG Haiou, LI Yahui   

  1. Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China
  • Received:2018-09-29 Online:2019-05-20 Published:2019-05-20

摘要:

为了阐明树鼩精子中是否存在丝氨酸/苏氨酸蛋白磷酸酶1γ2(PP1γ2)及其在附睾精子中的存在形式,进而探究PP1γ2对精子成熟和运动性的调控作用,本试验以树鼩为研究对象,采用Western blotting分析了不同条件下树鼩附睾头和附睾尾精子中PP1γ2的存在形式和磷酸化程度,探讨了双丁酰环腺苷酸(db-cAMP)、3-异丁基-1-甲基黄嘌呤(IBMX)或Ca2+对树鼩精子中PP1γ2磷酸化表达水平的影响,并进一步研究了磷酸酶抑制剂冈田酸(okadaic acid,OA)和花萼海绵诱癌素A (calyculin A,CA)对树鼩精子中PP1γ2磷酸化程度的影响及其对树鼩附睾头和附睾尾精子运动度的影响。结果显示,在树鼩附睾头和附睾尾精子中均存在PP1γ2,且在等量的附睾头和附睾尾精子蛋白中,PP1γ2在附睾尾精子的磷酸化程度远高于附睾头精子;db-cAMP、IBMX或Ca2+不改变PP1γ2的磷酸化水平;磷酸酶抑制剂OA和CA能明显提高附睾头和附睾尾中PP1γ2磷酸化的程度,且能显著提高精子(尤其是附睾头精子)的运动度(P<0.05),OA和CA的最佳作用浓度分别为1 μmol/L和10 nmol/L,最佳作用时间分别为15、20 min。本研究结果表明,蛋白磷酸酶PP1γ2对树鼩精子成熟及运动性具有重要的调控作用,其主要通过磷酸化和去磷酸化的变化发挥作用。

关键词: 树鼩; 丝氨酸/苏氨酸蛋白磷酸酶1γ 2(PP1γ 2); 精子成熟; 运动性; 附睾

Abstract:

In order to elucidate whether PP1γ2 is present or not in tree shrews'spermatozoa and its form in epididymal sperm,and further explore the regulation of PP1γ2 on sperm maturation and motility in tree shrews,the present form and phosphorylation levels of PP1γ2 in caput and cauda epididymal spermatozoa under various conditions have been analyzed by Western blotting.The effects of db-cAMP,IBMX or Ca2+ on the phosphorylation levels of PP1γ2 in tree shrew sperm were investigated.The effects of okadaic acid (OA) and calyculin A (CA) on the phosphorylation level of PP1γ2 and motility in caput and cauda epididymal spermatozoa were further studied with tree shrews as experimental animals in the present study.The results showed that PP1γ2 was present in caput and cauda epididymal spermatozoa of tree shrew,and the phosphorylation level of PP1γ2 was far higher in cauda epididymal spermatozoa than that of caput spermatozoa.db-cAMP,IBMX or Ca2+ did not change the phosphorylation level of PP1γ2,OA or CA could significantly increase not only the phosphorylation level of PP1γ2 in caput and cauda epididymal spermatozoa but also sperm motility (P<0.05),especially for caput epididymal spermatozoa.The optimal concentrations of OA and CA were 1 μmol/L and 10 nmol/L,respectively,and the optimal incubation time were 15 and 20 min,respectively.This study indicated that PP1γ2 played an important role in regulating sperm maturation and motility in tree shrews through phosphorylation and dephosphorylation.

Key words: tree shrew, PP1γ2, sperm maturation, motility, epididymis

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