Pax6Gene Expression Analysis By In Situ Hybridization During Lens Regeneration In Bufo Raddei Strauch Tadpole

We studied temporal and special expression of pax6gene in the process of lens regeneration of Bufo Raddei Strauch tadpole by gene cloning and in situ hybridization. The experimental results showed that during the early period of lens regeneration, pax6mRNA was detected in both dorsal and ventral iris depigment cell and proliferation cell, when regenerated lens cells differentiated, more pax6expressed in lens epithelium, at the same time, we found that the distribution of pax6mRNA in retina, cornea and lens epithelium, the expression in retina was stable, the gene resided mainly in Ganglion cells layer, inner nuclear layer, outer nuclear layer, as well as ciliary marginal zone. We preliminary identified the origin of the regenerated lens in Bufo Raddei Strauch tadpole and the changes of pax6during the process were closely related with lens regeneration.βB1crystallin is an important structural protein whose expression is regulated by Pax6,c-Maf and Proxl.Immunofluorescence was used to analyse the expression of βB1crystallin on day15,21after the lens removal in Bufo Raddei Strauch tadpole, the experimental results showed that (3B1crystallin was expressed both in regenerated and natraul lens.Our results further proved that the processes in Bufo Raddei Strauch tadpole lens regeneration included the dedifferentiation of dorsal iris pigmented cells, cell proliferation, and regenerated lens cells redifferentiation. The regenerated lens and original lens both expressed βB1crystallin.Comparing with the relevant studies in newt and Xenopus laevis, we found that Bufo Raddei Strauch tadpole and newt were relatively similar but with some differences during the process of lens regeneration, investigation of the causes that lead to these differences will thus shed new light for better understanding of the mechanisms of transdifferentiation in lens regeneration.