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Lim1 has been reported to regulate the head, nerve system and kidney development in mouse, zebrafish and Xenopus. In the present study, we demonstrate that Xlim1 plays a pivotal role in the pronephric specification and development in Xenopus. In Xenopus embryos at 4-cell and 8-cell stage, loss of function experiments using DEED depletion showed that down-regulation of Xlim1 expression severely impairs the formation of pronephros, assessed by pax8 expression. Overexpressing Xlim1 by injecting constitutively active Lim1 constructs in Xenopus embryos induce remarking enlargement of pronephros at various stages during development, assessed by both in situ hybridization and immunohistochemistry staining. Furthermore we studied the working window of Xlim1 on pronephric specification in mesoderm and observed the peak time point is at stage 10.5 while the whole working window is between stage 9 and 12.5.