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Ascidian is a useful experimental animal for studying body planning principles and host defense mechanisms employed by the phylum chordata. Toward this goal, genome and cDNA/EST projects of Ciona intestinalis have been undertaken. Using cDNAs and ESTs derived from Ciona hemocytes, we identified 79 possible hemocyte-preferential transcripts and determined the cDNA sequence of each clone. The amino acid sequence of each encoded polypeptide was predicted as well. Among these cDNAs, we identified three transcripts that may be involved in characteristic cell-cell communication in Ciona. These transcripts encoded leucine-rich repeat-containing RP105-like, IL-17 receptor/similar expression to FGF-like, and ectodysplasin-like polypeptide of the tunmr necrosis factor famlly, and they are expressed abundantly in hemocytes.

Bax and Bcl-XL are key regulators of apoptosis in mammals. Here we report the functional characterization of two Bcl-2 homologues, ciBax and ciBcl-XL, in a basal invertebrate-chordate ascidian Ciona intestinalis. CiBax is a Ciona homologue of the BH1-3 pro-apoptotic protein Bax, whereas ciBcl-XL is a Bcl-XL-like anti-apoptotic protein. Molecular modeling analysis showed that ciBax and ciBcl-XL share both sequence and structural similarities to human Bax and Bcl-XL, respectively. Like their human counterparts, ciBax could form a homodimer or oligomers as well as heterodimerize with ciBcl-XL, and overexpression of ciBax caused apoptosis that could be attenuated by ciBcl-XL. Mutagenesis studies showed that the BH3 domain of ciBax is critical for its cell death-inducing function and also for its interaction with ciBcl-XL. In Ciona embryos, ectopic expression of ciBax but not its BH3 deletion mutant resulted in cell dissociation and apoptosis after late gastrula stage of embryonic development. Moreover, not only wild type ciBcl-XL but also a mutant ciBcl-XL(F101V), which is unable to interact with ciBax, could block cell dissociation and developmental deficit in Ciona embryos induced by overexpression of ciBax. Taken together, these findings suggest that functional homologues of both the BH1-3 death effector Bax and the pro-survival Bcl-XL exist in sea squirt Ciona intestinalis, and they control the cell death machinery independent of their heterodimerization.