Intron retention facilitates splice variant diversity in calcium-activated big potassium channel populations
Intron retention facilitates splice variant diversity in calcium-activated big potassium channel populations
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DOI：
10.1073/pnas.1015264107
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发表时间：
2010-12-07
影响因子：
11.1
通讯作者：
Eberwine, James H.
中科院分区：
文献类型：
--
作者：
Bell, Thomas J.;Miyashiro, Kevin Y.;Eberwine, James H.
我们报告说，应力轴调节外显子（STREX）含钙激活大钾（BKCa）通道剪接变异体的表达和生理调节的部分细胞质剪接和内含子保留。NextGen对混合海马树突样本的mRNA互补序列进行测序，发现BKCa mRNA中的内含子17 a（i17 a），即紧接在STREX之前的内含子。对i17 a的进一步分子分析显示，大多数含有i17 a的BKCa通道mRNA与STREX相关。i17 a siRNA处理后，STREX蛋白免疫细胞化学显示含STREX的BKCa通道蛋白的水平降低和亚细胞分布改变。选择性减少i17 a-BKCa或STREX-BKCa mRNA诱导海马神经元爆发放电特性的类似变化。总的来说，这些数据表明，通过细胞质剪接和内含子保留的STREX剪接变体调节有助于在海马神经元中产生STREX依赖性BKCa电流多样性。
We report that the stress axis-regulated exon (STREX)-containing calcium-activated big potassium (BKCa) channel splice variant expression and physiology are regulated in part by cytoplasmic splicing and intron retention. NextGen sequencing of the mRNA complement of pooled hippocampal dendrite samples found intron 17a (i17a), the intron immediately preceding STREX, in the BKCa mRNA. Further molecular analyses of i17a revealed that the majority of i17a-containing BKCa channel mRNAs associate with STREX. i17a siRNA treatment followed by STREX protein immunocytochemistry demonstrated both reduced levels and altered subcellular distribution of STREX-containing BKCa channel protein. Selective reduction of i17a-BKCa or STREX-BKCa mRNAs induced similar changes in the burst firing properties of hippocampal neurons. Collectively, these data show that STREX splice variant regulation via cytoplasmic splicing and intron retention helps generate STREX-dependent BKCa current diversity in hippocampal neurons.