Rouhier Matthew F, Piermarini Peter M
Department of Entomology, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH 44691, USA.
Department of Entomology, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH 44691, USA.
Insect Biochem Mol Biol. 2014 May;48:91-9. doi: 10.1016/j.ibmb.2014.03.003. Epub 2014 Mar 18.
Inward-rectifier potassium (Kir) channels play key roles in nerve, muscle, and epithelial cells in mammals, but their physiological roles in insects remain to be determined. The yellow fever mosquito (Aedes aegypti) possesses five different genes encoding Kir channel subunits: Kir1, Kir2A, Kir2B, Kir2B', and Kir3. We have recently cloned and characterized the Kir1, Kir2B, and Kir3 cDNAs in the renal (Malpighian) tubules of adult female Ae. aegypti. Here we characterize the expression of the Kir2A gene in Ae. aegypti, which was not abundantly expressed in Malpighian tubules. We find that the 1) Kir2A gene is expressed primarily in the midgut and hindgut of adult female mosquitoes, and 2) Kir2A mRNAs are alternatively spliced into three distinct variants (Kir2A-a, -b, and -c). The deduced Kir2A proteins from these splice forms share a completely conserved transmembrane domain (a pore-forming domain flanked by two transmembrane-spanning segments), but possess novel NH2-terminal and/or COOH-terminal domains. Semi-quantitative RT-PCR analyses indicate that the splice variants exhibit both developmental- and tissue-specific expression. Lastly, we provide insights into the conservation of alternative splicing among the Kir2A genes of dipterans, which may add molecular diversity that compensates for the relatively limited number of Kir channel genes in insects compared to mammals.
内向整流钾(Kir)通道在哺乳动物的神经、肌肉和上皮细胞中发挥关键作用,但其在昆虫中的生理作用仍有待确定。黄热病蚊子(埃及伊蚊)拥有五个编码Kir通道亚基的不同基因:Kir1、Kir2A、Kir2B、Kir2B'和Kir3。我们最近克隆并鉴定了成年雌性埃及伊蚊肾(马氏)小管中的Kir1、Kir2B和Kir3 cDNA。在此,我们鉴定了埃及伊蚊中Kir2A基因的表达情况,该基因在马氏小管中表达不丰富。我们发现:1)Kir2A基因主要在成年雌性蚊子的中肠和后肠中表达;2)Kir2A mRNA可选择性剪接成三种不同的变体(Kir2A-a、-b和-c)。这些剪接形式推导的Kir2A蛋白共享一个完全保守的跨膜结构域(一个由两个跨膜片段侧翼的孔形成结构域),但具有新的NH2末端和/或COOH末端结构域。半定量RT-PCR分析表明,这些剪接变体表现出发育和组织特异性表达。最后,我们深入了解了双翅目昆虫Kir2A基因之间选择性剪接的保守性,这可能增加分子多样性,以弥补与哺乳动物相比昆虫中Kir通道基因数量相对有限的不足。