Gene-disruptions of C. elegans insulin-like genes Ceinsulin-1 and -2Tsuyoshi Kawano1, Naoya Kataoka1, Ikuko Tanaka1, Yasuo Kimura1, Shuji Honda2, Keiko Gengyo-Ando3 and Shohei Mitani31Department of Biological and Environmental Chemistry, Tottori University, 4-101, Minami, Koyama-cho, Tottori 680-8553, Japan2Tokyo Metropolitan Institute of Gerontology, 35-2, Sakae-cho, Itabashi-ku, Tokyo 173-0015, Japan3Department of Physiology, Tokyo Women , s Medical University School of Medicine, 8-1, Kawada-cho, Shinjuku-ku. Tokyo 162-8666, Japan In C. elegans, the insulin-signaling regulates the diapause, longevity and fat metabolism via the DAF-2, a homolog of insulin/IGF-I receptors. Our search for the DAF-2 ligands revealed two insulin/IGF-like peptides named Ceinsulin-1 and -2. We will also show phenotypes of the mutants lacking the genes.At first, Ceinsulin-1::gfp and Ceinsulin-2::gfp were constructed in a translational fusion manner and introduced into wild-type animals to search the cell expressing the Ceinsulin genes. The Ceinsulin-1::gfp is expressed at several neurons in the head and tail, whereas the Ceinsulin-2::gfp is expressed at several neurons only in the head. These cells do not overlap with chemosensory neurons filling DiI. Next, to Ceinsulin-1 and -2 deletion mutants designated
tm339 and
tm790, respectively, were generated and isolated with the TMP/UV method followed by the sib-screening. The
tm339 has deletion of approximately 1.4 kb covering two of three exons in the T28B8.2 gene. The
tm790 lays approximately deletion of 0.6 kb on two exons in the F56F3.6 gene. The
tm339 exhibits a slightly extended life span as our previous RNAi experiment, while the
tm790 shows almost the same life span as wild-type animals. Now, we are examining effects of these gene-disruptions on dauer larva formation.Gengyo-Ando, K and Mitani, S. (2000) Biochem. Biophys. Res. Commun. 269: 64-69.Kawano, T. et al. (2000) Biochem. Biophys. Res. Commun. 273: 431-436.Kawano, T. et al. (2003) Biosci. Biotechnol.Biochem. 67: 2678-2682.