title: Cloning, expression, and functional characterization of the rat Pax6 5a orthologous splicing variant.
abstract: 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.
authors: Department of Neurology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. Electronic address: fei.wei@gmail.com.
authors: Fei
authors: F
authors: Wei
authors: Reproductive Center & Gynecology Department of Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Min
authors: M
authors: Li
authors: Department of Neurology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors: Sai-Yu
authors: SY
authors: Cheng
authors: Trauma Center & Emergency Room of Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Liang
authors: L
authors: Wen
authors: Trauma Center & Emergency Room of Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Ming-Hua
authors: MH
authors: Liu
authors: Department of Neurology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors: Jie
authors: J
authors: Shuai
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D017931:DNA Primers; D005136:Eye Proteins / Q000235:genetics*; D057809:HEK293 Cells; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D051757:PAX5 Transcription Factor / Q000235:genetics*; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics*; D012326:RNA Splicing; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D005136:Eye Proteins; D018398:Homeodomain Proteins; D051757:PAX5 Transcription Factor; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; C000605733:Pax6 protein, rat; D012097:Repressor Proteins
keywords: Homeodomain; Mesenchymal stem cell; Paired domain; Pax6 isoform; Proliferation