Pax6 And Six3 Expression Analysis During Lens Regeneration In Xenopus Laevis

Objectives were study by immunofluorescence,Western Blot and qRT-PCR to compare the expression of Pax6 and Six3 in the process of lens regeneration of Xenopus laevis tadpole.The immunofluorescence results demonstrated that the lens regenerate after lentectomy,Pax6 expressed in cornea healed and thickened on 3 day after lentectomy;Pax6 expressed in cornea and new lens,Six3 expressed in outer cornea and new lens,the new lens on inner cornea adhesively on 7 day after lentectomy;new lens centre began to fibrosis and pulled away from the cornea,Pax6 expressed in outer cornea and Six3 expressed in outer cornea and lens epithelium on 14 day after lentectomy;lens fibrosis and began hyalinize on 21 day after lentectomy;new lens were basically completed,Pax6 and Six3 expressed in lens epithelium on 30 day after lentectomy.Pax6 and Six3 protein were expressed and the position stable in the retina after lentectomy,Pax6 protein were main expressed in Outer plexiform layer,Six3 protein were main expressed in Ganglia cell layer,Inner nuclear layer and Outer nuclear layer.The Western Blot results demonstrated that the expression of Pax6 compared with the control.There was extreme significant difference between 3 day,7 day,21 day,30 day and normal.The expression of Six3 compared with the control.There was significant difference between 0 day,30 day and normal.There was extreme significant difference between 3 day,21 day and normal.The qRT-PCR results demonstrated that Six3 mRNA were expressed in different periods after lentectomy.There was significant difference between 7 day and normal.There was extreme significant difference between 3 day,14 day,21 day,30 day and normal.The above experiment by histological staining and fluorescence imaging further proved that,Xenopus laevis len regeneration through the process of corneal epithelium cells dedifferentiation,capsule formation,lens fibrosis,and then form a transparent structure of the lens.The Western Blot and qRT-PCR results demonstrated that cell of outer cornea proliferation differentiation capsule by Pax6 and Six3 in initiation of regeneration,and these genes are closely associated with crystalline expression and cell fibrosis in later period.The study aims to reveal the expression of Pax6 and Six3 genes in Xenopus laevis lens regeneration,in order to provide the theory evidence of amphibians lens regeneration.