Department of Neurology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
Reproductive Center & Gynecology Department of Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Gene. 2014 Aug 15;547(1):169-74. doi: 10.1016/j.gene.2014.06.032. Epub 2014 Jun 18.
Pax6 functions as a pleiotropic regulator in eye development and neurogenesis. Its splice variant Pax6 5a has been cloned in many vertebrate species including human and mouse, but never in rat. This study focused on the cloning and characterization of the Pax6 5a orthologous splicing variant in rat. It was cloned from Sprague-Dawley rats 10 days post coitum (E10) by RT-PCR and was sequenced for comparison with Pax6 sequences in the GenBank by BLAST. The rat Pax6 5a was revealed to contain an additional 42 bp insertion at the paired domain. At the nucleotide level, the rat Pax6 5a coding sequence (1,311 bp) had a higher degree of homology to the mouse (96% identical) than to the human (93% identical) sequence. At the amino acid (aa) level, rat PAX6 5a shares 99.8% identity with the mouse sequence and 99.5% with the human sequence. The splice variant is preferentially expressed in the rat E10 embryonic headfolds and not in the trunk of neurula. Its effects on the proliferation of rat mesenchymal stem cells (rMSCs) were preliminarily evaluated by the MTT assay. Both pLEGFP-Pax6 5a-transfected cells and pLEGFP-Pax6-transfected cells exhibited a similar growth curve (P>0.05), suggesting that the Pax6 5a has a similar effect on the proliferation of rMSCs as Pax6.
Pax6 作为一种多效调节因子在眼睛发育和神经发生中发挥作用。其剪接变异体 Pax6 5a 已在包括人类和小鼠在内的许多脊椎动物物种中被克隆，但从未在大鼠中被克隆。本研究专注于克隆和鉴定大鼠 Pax6 5a 同源剪接变异体。该研究通过 RT-PCR 从 Sprague-Dawley 大鼠胚胎 10 天（E10）中克隆，并与 GenBank 中的 Pax6 序列进行 BLAST 比较以确定其序列。研究揭示大鼠 Pax6 5a 在配对结构域处含有额外的 42bp 插入。在核苷酸水平上，大鼠 Pax6 5a 编码序列（1311bp）与小鼠（96%相同）的同源性高于与人（93%相同）的序列。在氨基酸（aa）水平上，大鼠 PAX6 5a 与小鼠序列共享 99.8%的同一性，与人类序列共享 99.5%的同一性。该剪接变异体在大鼠 E10 胚胎头褶中优先表达，而不在神经胚的躯干中表达。通过 MTT 测定法初步评估了该剪接变异体对大鼠间充质干细胞（rMSCs）增殖的影响。pLEGFP-Pax6 5a 转染细胞和 pLEGFP-Pax6 转染细胞均表现出相似的生长曲线（P>0.05），这表明 Pax6 5a 对 rMSCs 的增殖具有与 Pax6 相似的作用。
