Inositol 1,4,5-trisphosphate receptors (IP3Rs) are intracellular Ca2+ channels that elevate cytoplasmic Ca2+ in response to the second messenger IP3. Here, we describe the identification and in vivo functional characterization of the planarian IP3R, the first intracellular Ca2+ channel to be defined in flatworms. A single IP3R gene in Dugesia japonica encoded a 2666 amino acid protein (Dj.IP3R) that shared well conserved structural features with vertebrate IP3R counterparts. Expression of an NH2-terminal Dj.IP3R region (amino acid residues 223-585) recovered high affinity 3H-IP3 binding (0.9±0.1nM) which was abolished by a single point mutation of an arginine residue (R495L) important for IP3 coordination. In situ hybridization revealed that Dj.IP3R mRNA was most strongly expressed in the pharynx and optical nerve system as well as the reproductive system in sexualized planarians. Consistent with this observed tissue distribution, in vivo RNAi of Dj.IP3R resulted in a decreased egg-laying behavior suggesting Dj.IP3R plays an upstream role in planarian reproductive physiology.
Bibliographical noteFunding Information:
Work in the laboratory was supported by the NIH ( GM088790 to J.S.M.) and NSF (MCB 0919933 ). J.D.C. was supported by a Stem Cell Biology Training Grant ( T32HD060536 ). Appendix A