High cGMP synthetic activity in carp cones

• DOI: 10.1073/pnas.0812781106
• Publication Date: 2009-06-26T01:59:42Z
• Authors: Norihiko Takemoto; Shuji Tachibanaki; Satoru Kawamura
• Keywords: 0303 health sciences; Carps; Base Sequence; Adaptation, Ocular; Reverse Transcriptase Polymerase Chain Reaction; Immunoblotting; Molecular Sequence Data; Sequence Analysis, DNA; Guanylate Cyclase-Activating Proteins; Electrophysiology; Kinetics; 03 medical and health sciences; Guanylate Cyclase; Retinal Cone Photoreceptor Cells; Animals; Cyclic GMP; DNA Primers

Cones show briefer light responses than rods and do not saturate even under very bright light. Using purified rod and cone homogenates, we measured the activity of guanylate cyclase (GC), an enzyme responsible for cGMP synthesis and therefore recovery of a light response. The basal GC activity was 36 times higher in cones than in rods: It was mainly caused by higher expression levels of GC in cones (GC-C) than in rods (GC-R). With identification and quantification of GC-activating protein (GCAP) subtypes expressed in rods and cones together with determination of kinetic parameters of GC activation in the presence and absence of GCAP, we estimated the in situ GC activity in rods and cones at low and high Ca 2+ concentrations. It was revealed that the GC activity would be >10 times higher in cones than in rods in both the dark-adapted and the light-adapted states. Electrophysiological estimation of the GC activity measured in the truncated preparations of rod and cone outer segments gave consistent results. Our estimation of the in situ GC activity reasonably explained the rapid recovery and nonsaturating behavior of cone light responses.