A small scale expression screen identifies tissue specific markers in the Dugesia japonica strain Pek-1

Freshwater planaria has tremendous capacity to reform the missing part of the body and therefore is considered as one of the most important model organism for regeneration study. At present, Schmidtea mediterranea and Dugesia japonica are the two major species utilized for laboratory manipulations. Dugesia japonica flatworms are widely distributed in the Far East including Cherry Valley region in the north-west area of Beijing, China. We reported here the establishment of an asexual Dugesiajaponica strain Pek-1, as a suitable system for regeneration study. Using morphological, karyotypical as well as phylogenetic analyses, we confirmed that these flatworms indeed belonged to Dugesia japonica. We went on to show that the commonly used in situ probes and immunohistochemistry reagents and protocols were applicable to the Pek-1 strain. Using this strain, we carried out small scale analysis on EST, RNAi and gene expression. We identified 193 unique EST sequences and 65 of them had not been reported in planarian. By RNAi analysis, we showed that 48 genes, when down-regulated individually, had no effect on regeneration. Furthermore, we identified 3 groups of tissue specific expressing genes that were useful for cell lineage analysis. We concluded that the Dugesiajaponica Pek-1 strain could be another suitable animal model to regeneration research.

Djrho2 is involved in regeneration of visual nerves in Dugesia japonica

The freshwater planarian is a powerful animal model for studying regeneration and stem cell activity in vivo. During regeneration, stem cells （neoblasts in planarian） migrated to the wounding edge to re-build missing parts of the body. However, proteins involved in regulating cell migration during planarian regeneration have not been studied extensively. Here we report two small GTPase genes （Djrho2 and Djrho3） of Dugesiajaponica （strain Pek-1）. In situ hybridization results indicated that Djrho2 was expressed throughout the body with the exception of thc pharynx region while Djrho3 was specifically expressed along the gastro-vascular system. Djrho2 was largely expressed in neoblasts since its expression was sensitive to X-ray irradiation. In Djrho2-RNAi planarians, smaller anterior blastemas were observed in tail fragments during regeneration. Consistently, defective regeneration of visual nerve was detected by immunostairming with VC-1 antibody. These results suggested that Djrho2 is required for proper anterior regeneration in planairan. In contrast, no abnormality was observed after RNAi of Djrho3. We compared protein compositions of control and Djrho2-RNAi planarians using an optimized proteomic approach. Twenty-two up-regulated and 26 de-regulated protein spots were observed in the two-dimensional elec- trophoresis gels, and 17 proteins were successfully identified by Mass Spectrometry （MS） analysis. Among them, 6 actin-binding or cytoskeleton-related proteins were found de-expressed in Djrho2-RNAi animals, suggesting that abnormal cytoskeleton assembling and cell migration were likely reasons of defected regeneration.