title: Inositol-1,4,5-trisphosphate receptors in the vertebrate retina.
abstract: Evidence has shown an activation of phosphatidylinositol 4,5-bisphosphate (PIP2) specific phospholipase C (PtdIns-PLC) by light in the vertebrate retina and rod outer segments (ROS), suggesting important roles for its two metabolites, 1,2-diacylglycerol (DG) and inositol-1,4,5-trisphosphate [Ins(1,4,5)P3]. DG activates protein kinase C (PKC) and Ins(1,4,5)P3 releases bound intracellular calcium. Since Ca2+ plays an important role in light adaptation, the presence of Ins(1,4,5)P3 receptors in ROS may indicate a regulatory role of Ins(1,4,5)P3 to the free Ca2+ content. In the present study, we investigated the Ins(1,4,5)P3 receptors in whole retinal membranes and several subcellular fractions prepared from bovine retinas. Scatchard analyses of binding data for retinal membrane preparations showed a single, high-affinity binding site with equilibrium dissociation constant (Kd) of 24 +/- 2 nM and maximal binding capacity (Bmax) of 353 +/- 15 fmol/mg protein at pH 7.4. Specific binding was found in both small and large synaptosomal preparations representing inner and outer plexiform layers, respectively. A detectable, but low abundance of Ins(1,4,5)P3-specific binding in ROS was observed at both pH 7.4 and 8.3, but no specific binding of Ins(1,4,5)P3 was found in isolated outer segment discs. The binding of Ins(1,4,5)P3 in ROS was reduced by addition of ATP, suggesting a regulatory role for this nucleotide. Addition of calcium, sodium, and potassium ions also reduced specific binding of Ins(1,4,5)P3. Immunocytochemical studies indicate intense staining in the inner segment and extending to the ROS. Inner and outer plexiform layers were also stained. These findings show that the Ins(1,4,5)P3 receptor is present in photoreceptor cells and inner and outer plexiform layers in the vertebrate retina.
authors.affiliation: Cullen Eye Institute, Baylor College of Medicine, Houston, TX 77030.
authors.forename: N S
authors.initials: NS
authors.lastname: Day
authors.forename: C A
authors.initials: CA
authors.lastname: Koutz
authors.forename: R E
authors.initials: RE
authors.lastname: Anderson
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D015220:Calcium Channels / Q000378:metabolism*; D002417:Cattle; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D053496:Inositol 1,4,5-Trisphosphate Receptors; D010786:Photoreceptor Cells / Q000378:metabolism; D011817:Rabbits; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D012160:Retina / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism
chemical_list: D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D018160:Receptors, Cytoplasmic and Nuclear; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate