Whitney Irene E, Kautzman Amanda G, Reese Benjamin E
Neuroscience Research Institute, University of California at Santa Barbara, Santa Barbara, CA 93106-5060, United States; Department of Molecular, Cellular and Developmental Biology, University of California at Santa Barbara, Santa Barbara, CA 93106-9625, United States.
Neuroscience Research Institute, University of California at Santa Barbara, Santa Barbara, CA 93106-5060, United States; Department of Psychological & Brain Sciences, University of California at Santa Barbara, Santa Barbara, CA 93106-9660, United States.
Mol Cell Neurosci. 2015 Mar;65:102-13. doi: 10.1016/j.mcn.2015.03.006. Epub 2015 Mar 6.
Islet-1 (Isl1) is a LIM-homeodomain (LIM-HD) transcription factor that functions in a combinatorial manner with other LIM-HD proteins to direct the differentiation of distinct cell types within the central nervous system and many other tissues. A study of pancreatic cell lines showed that Isl1 is alternatively spliced generating a second isoform, Isl1β, which is missing 23 amino acids within the C-terminal region. This study examines the expression of the canonical and alternative Isl1 transcripts across other tissues, in particular, within the retina, where Isl1 is required for the differentiation of multiple neuronal cell types. The alternative splicing of Isl1 is shown to occur in multiple tissues, but the relative abundance of Isl1α and Isl1β expression varies greatly across them. In most tissues, Isl1α is the more abundant transcript, but in others the transcripts are expressed equally, or the alternative splice variant is dominant. Within the retina, differential expression of the two Isl1 transcripts increases as a function of development, with dynamic changes in expression peaking at E16.5 and again at P10. At the cellular level, individual retinal ganglion cells vary in their expression, with a subset of small-to-medium sized cells expressing only the alternative isoform. The functional significance of the difference in protein sequence between the two Isl1 isoforms was also assessed using a luciferase assay, demonstrating that the alternative isoform forms a less effective transcriptional complex for activating gene expression. These results demonstrate the differential presence of the canonical and alternative isoforms of Isl1 amongst retinal ganglion cell classes. As Isl1 participates in the differentiation of multiple cell types within the CNS, the present results support a role for alternative splicing in the establishment of cellular diversity in the developing nervous system.
胰岛-1(Isl1)是一种含LIM结构域的同源异型转录因子(LIM-HD),它与其他LIM-HD蛋白以组合方式发挥作用,指导中枢神经系统和许多其他组织中不同细胞类型的分化。一项对胰腺细胞系的研究表明,Isl1存在可变剪接,产生第二种异构体Isl1β,其在C末端区域缺失23个氨基酸。本研究检测了标准型和可变型Isl1转录本在其他组织中的表达情况,特别是在视网膜中,多种神经元细胞类型的分化需要Isl1。结果表明,Isl1的可变剪接发生在多种组织中,但Isl1α和Isl1β表达的相对丰度在不同组织中差异很大。在大多数组织中,Isl1α是更丰富的转录本,但在其他组织中,两种转录本表达量相当,或者可变剪接变体占主导。在视网膜内,两种Isl1转录本的差异表达随着发育而增加,表达的动态变化在胚胎第16.5天和出生后第10天达到峰值。在细胞水平上,单个视网膜神经节细胞的表达存在差异,一部分中小尺寸细胞只表达可变异构体。还使用荧光素酶测定法评估了两种Isl1异构体之间蛋白质序列差异的功能意义,结果表明可变异构体形成的激活基因表达的转录复合物效率较低。这些结果证明了Isl1的标准型和可变型异构体在视网膜神经节细胞类别中的差异存在。由于Isl1参与中枢神经系统中多种细胞类型的分化,目前的结果支持可变剪接在发育中的神经系统细胞多样性建立中的作用。