Departments of Ophthalmology and Physiology, School of Medicine, University of California, San Francisco, San Francisco, California 94143, Albert Eye Research Institute, Duke University Medical Center, Durham, North Carolina 27710, and Department of Biology, University of Oregon, Eugene, Oregon 97403.
J Neurosci. 2013 Nov 6;33(45):17763-76. doi: 10.1523/JNEUROSCI.2659-13.2013.
The ligand sensitivity of cGMP-gated (CNG) ion channels in cone photoreceptors is modulated by CNG-modulin, a Ca(2+)-binding protein. We investigated the functional role of CNG-modulin in phototransduction in vivo in morpholino-mediated gene knockdown zebrafish. Through comparative genomic analysis, we identified the orthologue gene of CNG-modulin in zebrafish, eml1, an ancient gene present in the genome of all vertebrates sequenced to date. We compare the photoresponses of wild-type cones with those of cones that do not express the EML1 protein. In the absence of EML1, dark-adapted cones are ∼5.3-fold more light sensitive than wild-type cones. Previous qualitative studies in several nonmammalian species have shown that immediately after the onset of continuous illumination, cones are less light sensitive than in darkness, but sensitivity then recovers over the following 15-20 s. We characterize light sensitivity recovery in continuously illuminated wild-type zebrafish cones and demonstrate that sensitivity recovery does not occur in the absence of EML1.
在视锥细胞中,cGMP 门控(CNG)离子通道的配体敏感性受 CNG-调节蛋白(一种 Ca2+结合蛋白)调节。我们通过在斑马鱼中进行基于 morpholino 的基因敲低实验,研究了 CNG-调节蛋白在体内光转导中的功能作用。通过比较基因组分析,我们在斑马鱼中鉴定出了 CNG-调节蛋白的同源基因 eml1,这是一个在迄今为止所有测序的脊椎动物基因组中都存在的古老基因。我们比较了野生型视锥细胞和不表达 EML1 蛋白的视锥细胞的光反应。在缺乏 EML1 的情况下,暗适应的视锥细胞比野生型视锥细胞对光的敏感约 5.3 倍。之前在几种非哺乳动物物种中的定性研究表明,在连续光照开始后,视锥细胞对光的敏感性比在黑暗中要低,但在接下来的 15-20 秒内,敏感性会恢复。我们描述了在连续光照下野生型斑马鱼视锥细胞的光敏感性恢复,并证明在缺乏 EML1 的情况下,敏感性恢复不会发生。