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Expression of the Third Component of Complement, C3, in Regenerating Limb Blastema Cells of Urodeles^{1 ,2}

Katia Del Rio-Tsonis^*, Panagiotis A. Tsonis^*, Ioannis K. Zarkadis^3,, Andreas G. Tsagas and John D. Lambris^4,

^* Laboratory of Molecular Biology, Department of Biology, University of Dayton, Dayton, OH 45469; and Protein Chemistry Laboratory, Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104

In this study we have shown that complement component C3 is expressed in the regenerating tissue during urodele limb regeneration. C3 was expressed in the dedifferentiated regeneration blastema and in the redifferentiated limb tissues in the axolotl, Amblystoma mexicanum, and in Notophthalmus viridescens. This expression was verified by immunofluorescent staining using an Ab against axolotl C3 and by in situ hybridization with an axolotl C3 cDNA probe. In the early stages of regeneration C3 appeared to be equally present in all mesenchymal cells and in the wound epithelium, whereas in the later stages it was mainly expressed in the differentiating muscle cells. Since no expression was seen in the developing limb, it appears that the C3 expression was specific to the regeneration process. We then demonstrated by hybridization experiments that a blastema cell line of myogenic origin expresses C3. All these findings implicate C3 in the dedifferentiation process and may indicate a new role for this molecule in muscle differentiation.

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