Molecular Characterization and Tissue Distribution of Mitochondrial Ca2+ -Dependent Solute Carrier Protein during Prevention of Diapause by HCl Treatment in the Silkworm, Bombyx mori
Molecular Characterization and Tissue Distribution of Mitochondrial Ca2+ -Dependent Solute Carrier Protein during Prevention of Diapause by HCl Treatment in the Silkworm, Bombyx mori
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DOI：
10.2108/zs180041
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发表时间：
2018-12
影响因子：
0.9
通讯作者：
Ryo Kitta;Y. Yamahama;Takayuki Yamamoto;K. Mase;H. Sawada
中科院分区：
文献类型：
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作者：
Ryo Kitta;Y. Yamahama;Takayuki Yamamoto;K. Mase;H. Sawada
为阐明盐酸处理防止家蚕进入滞育的分子机制，对盐酸处理滞育卵的线粒体Ca 2+依赖性溶质载体蛋白（MCSC）进行了生化分析。我们以前的研究表明，盐酸处理引起Ca ~（2+）从滞育卵中流出。因此，我们试图分析MCSC，已知其与Ca 2+相关。将分离的B. MoriMCSC（BmMCSC）具有一个667个氨基酸的开放阅读框架（ORF），包含2个EF-手形钙结合结构域和3个线粒体溶质载体超家族的特征。盐酸处理后BmMCSC基因表达水平增加。Ca ~（2+）结合实验表明，重组BmMCSC（rBmMCSC）对Ca ~（2+）具有亲和力。用抗BmMCSC的免疫组化技术研究了滞育卵和盐酸处理的滞育卵中BmMCSC的分布。BmMCSC定位于两个卵的浆膜细胞中。这些结果提示，BmMCSC是由胞内Ca ~（2+）激活或被HCl处理后外排Ca ~（2+）激活的，可能参与了家蚕人工滞育预防或解除滞育的分子机制。
To clarify the molecular mechanism of prevention of entry into diapause in Bombyx mori by HCl treatment, we biochemically analyzed mitochondrial Ca2+ -dependent solute carrier protein (MCSC) in diapause eggs treated with HCl solution. Our previous studies revealed that HCl treatment causes Ca2+ to efflux from diapause eggs. Therefore, we attempted to analyze MCSC, which is known to associate with Ca2+ . The isolated cDNA of B. mori MCSC (BmMCSC) had an open reading flame (ORF) of 667 amino acid residues, and the ORF contained two EF-hand calcium-binding domains and three characteristic features of the mitochondrial solute carrier superfamily. The gene expression level of BmMCSC increased by HCl treatment. A Ca2+ binding assay indicated that recombinant BmMCSC (rBmMCSC) shows an affinity with Ca2 + . The distribution of BmMCSC was investigated with an immunohistochemical technique using antisera against BmMCSC in diapause eggs and HCl-treated diapause eggs. BmMCSC was localized in serosa cells in both eggs. These data may suggest that BmMCSC is activated by intracellular Ca2+ or efflux Ca2+ by HCl treatment, and that it plays a role in the molecular mechanisms of artificial diapause prevention or the breaking of diapause in the silkworm.