Plasma membrane calcium ATPase 4 (PMCA4) co-ordinates calcium and nitric oxide signaling in regulating murine sperm functional activity.
Plasma membrane calcium ATPase 4 (PMCA4) co-ordinates calcium and nitric oxide signaling in regulating murine sperm functional activity.
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DOI：
10.1002/jcp.25882
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发表时间：
2018-01
影响因子：
5.6
通讯作者：
Martin-DeLeon PA
中科院分区：
文献类型：
--
作者：
Olli KE;Li K;Galileo DS;Martin-DeLeon PA
精子活力降低（弱精子症）和不育是由质膜Ca2+- atp酶4 （Pmca4）基因的缺失引起的，该基因编码高度保守的Ca2+外排泵Pmca4。这是小鼠精子中主要的Ca2+清除蛋白。由于弱精子症中PMCA4缺失或活性降低的机制尚不清楚，我们研究了精子PMCA4是否负调控一氧化氮合酶（nos），以及当NO缺失时，过氧亚硝酸盐和氧化应激水平是否升高。利用共免疫沉淀（Co-IP）和荧光共振能量转移（FRET），我们发现PMCA4与被激活和被Ca2+离子载体处理的精子中升高的细胞质[Ca2+]中的nos以及与基础[Ca2+]（无激活精子）中的神经元（nNOS）存在关联。PMCA4-eNOS在失能和失能精子中的FRET效率分别为35%和23%，差异极显著（P<0.01），且分子间距<10 nm。对于PMCA4-nNOS，这种相互作用仅在有能力精子中出现，其FRET效率为24%，显著（P<0.05）高于无能力精子（6%）。PMCA4和nos被鉴定为包括Caveolin1在内的四元配合物的相互作用伙伴，该配合物以Ca2+依赖的方式与eNOS共同免疫沉淀。在Pmca4−/−精子中，NOS活性在失能/失能精子中升高2倍（与野生型相比），同时过氧亚硝酸盐水平升高2倍，凋亡生殖细胞数量显著增加（P<0.001）。这些数据支持一个四元复合物模型，其中PMCA4协调Ca2+和NO信号来维持运动，NO水平升高导致PMCA4−/−雄性弱精子症。他们认为人类弱精子症与PMCA4突变有关，具有诊断意义。
Reduced sperm motility (asthenospermia) and resulting infertility arise from deletion of the Plasma Membrane Ca2+-ATPase 4 (Pmca4) gene which encodes the highly conserved Ca2+ efflux pump, PMCA4. This is the major Ca2+ clearance protein in murine sperm. Since the mechanism underlying asthenospermia in PMCA4’s absence or reduced activity is unknown, we investigated if sperm PMCA4 negatively regulates nitric oxide synthases (NOSs) and when absent NO, peroxynitrite, and oxidative stress levels are increased. Using co-immunoprecipitation (Co-IP) and Fluorescence Resonance Energy Transfer (FRET), we show an association of PMCA4 with the NOSs in elevated cytosolic [Ca2+] in capacitated and Ca2+ ionophore-treated sperm and with neuronal (nNOS) at basal [Ca2+] (ucapacitated sperm). FRET efficiencies for PMCA4-eNOS were 35% and 23% in capacitated and uncapacitated sperm, significantly (P<0.01) different, with the molecules being <10 nm apart. For PMCA4-nNOS, this interaction was seen only for capacitated sperm where FRET efficiency was 24%, significantly (P<0.05) higher than in uncapacitated sperm (6%). PMCA4 and the NOSs were identified as interacting partners in a quaternary complex that includes Caveolin1, which co-immunoprecipitated with eNOS in a Ca2+-dependent manner. In Pmca4−/− sperm NOS activity was elevated 2-fold in capacitated/uncapacitated sperm (versus wild-type), accompanied by a 2-fold increase in peroxynitrite levels and significantly (P<0.001) increased numbers of apoptotic germ cells. The data support a quaternary complex model in which PMCA4 co-ordinates Ca2+ and NO signaling to maintain motility, with increased NO levels resulting in asthenospermia in Pmca4−/− males. They suggest the involvement of PMCA4 mutations in human asthenospermia, with diagnostic relevance.
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DOI：
10.1530/rep-09-0134
发表时间：
2009-09
期刊：
Reproduction (Cambridge, England)
影响因子：
--
作者：
Costello S;Michelangeli F;Nash K;Lefievre L;Morris J;Machado-Oliveira G;Barratt C;Kirkman-Brown J;Publicover S
通讯作者：
Publicover S
影响因子：
4
作者：
Herrero, MB;de Lamirande, E;Gagnon, C
通讯作者：
Gagnon, C
影响因子：
4.8
作者：
Ju, H;Zou, R;Venema, RC
通讯作者：
Venema, RC
影响因子：
6.7
作者：
O'Bryan, MK;Zini, A;Schlegel, PN
通讯作者：
Schlegel, PN
影响因子：
3.9
作者：
Oceandy, D.;Stanley, P. J.;Neyses, L.
通讯作者：
Neyses, L.