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)P3releases bound intracellular calcium. Since Ca2+plays an important role in light adaptation, the presence of Ins(1,4,5)P3receptors in ROS may indicate a regulatory role of Ins(1,4,5)P3to the free Ca2+content. In the present study, we investigated the Ins(1,4,5)P3receptors 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)P3was found in isolated outer segment discs. The binding of Ins(1,4,5)P3in 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)P3receptor is present in photoreceptor cells and inner and outer plexiform layers in the vertebrate retina.