Department of Biology, The University of Iowa, Iowa City, IA 52242, USA.
Proc Natl Acad Sci U S A. 2013 Jan 2;110(1):181-6. doi: 10.1073/pnas.1208866110. Epub 2012 Dec 17.
Ion homeostasis is a fundamental cellular process particularly important in excitable cell activities such as hearing. It relies on the Na(+)/K(+) ATPase (also referred to as the Na pump), which is composed of a catalytic α subunit and a β subunit required for its transport to the plasma membrane and for regulating its activity. We show that α and β subunits are expressed in Johnston's organ (JO), the Drosophila auditory organ. We knocked down expression of α subunits (ATPα and α-like) and β subunits (nrv1, nrv2, and nrv3) individually in JO with UAS/Gal4-mediated RNAi. ATPα shows elevated expression in the ablumenal membrane of scolopale cells, which enwrap JO neuronal dendrites in endolymph-like compartments. Knocking down ATPα, but not α-like, in the entire JO or only in scolopale cells using specific drivers, resulted in complete deafness. Among β subunits, nrv2 is expressed in scolopale cells and nrv3 in JO neurons. Knocking down nrv2 in scolopale cells blocked Nrv2 expression, reduced ATPα expression in the scolopale cells, and caused almost complete deafness. Furthermore, knockdown of either nrv2 or ATPα specifically in scolopale cells causes abnormal, electron-dense material accumulation in the scolopale space. Similarly, nrv3 functions in JO but not in scolopale cells, suggesting neuron specificity that parallels nrv2 scolopale cell-specific support of the catalytic ATPα. Our studies provide an amenable model to investigate generation of endolymph-like extracellular compartments.
离子稳态是一种基本的细胞过程,在听觉等兴奋细胞活动中尤为重要。它依赖于 Na(+)/K(+)ATP 酶(也称为 Na 泵),该酶由一个催化 α 亚基和一个 β 亚基组成,β 亚基对于其向质膜的转运和调节其活性是必需的。我们表明,α 和 β 亚基在果蝇听觉器官 Johnston's organ(JO)中表达。我们使用 UAS/Gal4 介导的 RNAi 在 JO 中单独敲低 α 亚基(ATPα 和 α-like)和 β 亚基(nrv1、nrv2 和 nrv3)的表达。ATPα 在包裹 JO 神经元树突的栉板细胞的腔外膜中表达升高,位于类似于内淋巴的隔室中。使用特定的驱动子在整个 JO 或仅在栉板细胞中敲低 ATPα,但不是 α-like,导致完全失聪。在 β 亚基中,nrv2 在栉板细胞中表达,nrv3 在 JO 神经元中表达。在栉板细胞中敲低 nrv2 会阻断 Nrv2 的表达,降低栉板细胞中 ATPα 的表达,并导致几乎完全失聪。此外,特异性敲低栉板细胞中的 nrv2 或 ATPα 会导致栉板空间中异常的、电子致密物质积累。同样,nrv3 在 JO 中发挥作用,但不在栉板细胞中,这表明神经元特异性与 nrv2 在栉板细胞中特异性支持催化 ATPα 相平行。我们的研究提供了一个可行的模型来研究内淋巴样细胞外隔室的生成。