High cGMP synthetic activity in carp cones.
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.
Takemoto
Norihiko
N
Graduate School of Frontier Biosciences, Graduate School of Science, Osaka University, Suita, Osaka 565-0871, Japan.
Tachibanaki
Shuji
S
Kawamura
Satoru
S
D000221:Adaptation, Ocular / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000378:metabolism*; D006152:Cyclic GMP / Q000096:biosynthesis*; D017931:DNA Primers / Q000235:genetics; D004594:Electrophysiology; D006162:Guanylate Cyclase / Q000378:metabolism*; D051604:Guanylate Cyclase-Activating Proteins / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D007700:Kinetics; D008969:Molecular Sequence Data; D017949:Retinal Cone Photoreceptor Cells / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D051604:Guanylate Cyclase-Activating Proteins; D006162:Guanylate Cyclase; D006152:Cyclic GMP