Cyclic AMP and glycogen synthase kinase 3 form a regulatory loop in spermatozoa.
Cyclic AMP and glycogen synthase kinase 3 form a regulatory loop in spermatozoa.
复制标题
DOI：
10.1002/jcp.26557
复制
发表时间：
2018-09
影响因子：
5.6
通讯作者：
Vijayaraghavan S
中科院分区：
文献类型：
--
作者：
Dey S;Goswami S;Eisa A;Bhattacharjee R;Brothag C;Kline D;Vijayaraghavan S
多方面糖原合成酶激酶(GSK3)在精子和男性生育中起着至关重要的作用。鉴于环磷酸腺苷(CAMP)在精子功能中起重要作用，我们研究了GSK3和cAMP途径是否可能相互关联。我们使用GSK3和可溶性腺酰环化酶(SAC)基因敲除小鼠和药物调节剂来研究这种关系。缺乏Gsk3α或Gsk3β的精子细胞内cAMP水平显著降低。当精子细胞被GSK3抑制剂SB216763处理时，也观察到了类似的结果。这种cAMP水平的降低不是由于对精子腺酰环化酶的影响，而是由于磷酸二酯酶(PDE)活性的升高。PDE4抑制剂RS25344或常规PDE抑制剂IBMX均可使缺乏GSK3、α或β异构体的精子的cAMP水平恢复。PDE活性测定还表明，在Gsk3α缺失精子中，过度激活的PDE4有助于降低cAMP水平，这表明在野生型精子中，PDE4的活性受到Gsk3的抑制。相反，由cAMP触发的PKA通过增加其磷酸化来影响GSK3的活性。抑制精子特异性蛋白磷酸酶1，γ-2，也可增加γ-3的磷酸化。用重组Pp1γ2进行下拉实验表明，PkA、Gsk3和Pp1γ2可能以复合体的形式存在。GSK3在成熟精子中的药理抑制导致卵子的体外受精显著减少。我们的结果表明，cAMP、PKA和GSK3在精子功能的调节中相互关联。
The multifaceted glycogen synthase kinase (GSK3) has an essential role in sperm and male fertility. Since cyclic AMP (cAMP) plays an important role in sperm function, we investigated whether GSK3 and cAMP pathways may be interrelated. We used GSK3 and soluble adenylyl cyclase (sAC) knockout mice and pharmacological modulators to examine this relationship. Intracellular cAMP levels were found to be significantly lower in sperm lacking GSK3α or GSK3β. A similar outcome was observed when sperm cells were treated with SB216763, a GSK3 inhibitor. This reduction of cAMP levels was not due to an effect on sperm adenylyl cyclase but was caused by elevated phosphodiesterase (PDE) activity. The PDE4 inhibitor RS25344 or the general PDE inhibitor IBMX could restore cAMP levels in sperm lacking GSK3α or β-isoform. PDE activity assay also showed that hyperactivated PDE4 contributes in lowering of cAMP levels in GSK3α null sperm suggesting that in wild-type sperm PDE4 activity is kept in check by GSK3. Conversely, PKA being triggered by cAMP, affected GSK3 activity through increasing its phosphorylation. Increased GSK3 phosphorylation also occurred by inhibition of sperm specific protein phosphatase type 1, PP1γ2. The relationship between cAMP, GSK3, and PP1γ2 activities was also confirmed in sperm from sAC null mice. Pull-down assay using recombinant PP1γ2 indicated that PKA, GSK3, and PP1γ2 could exist as a complex. Pharmacological inhibition of GSK3 in mature spermatozoa resulted in significantly reduced fertilization of eggs in vitro. Our results show that cAMP, PKA, and GSK3 are interrelated in regulation of sperm function.
登录
查看更多内容
DOI：
10.1038/nrd4228
发表时间：
2014-04
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1073/pnas.0400050101
发表时间：
2004-03-02
影响因子：
11.1
作者：
Esposito, G;Jaiswal, BS;Gossen, JA
通讯作者：
Gossen, JA
DOI：
10.1073/pnas.220413597
发表时间：
2000-10-24
影响因子：
11.1
作者：
Fang, XJ;Yu, SX;Mills, GB
通讯作者：
Mills, GB
影响因子：
4.4
作者：
Aquila, Saveria;Guido, Carmela;Ando, Sebastiano
通讯作者：
Ando, Sebastiano
影响因子：
5.8
作者：
Aquila, S;Gentile, M;Andò, S
通讯作者：
Andò, S