Abstract

We have used a preparation of isolated, intact rod photoreceptors to correlate the effects of flash illumination on the intracellular cyclic GMP content and the membrane current. We find that the recovery of cyclic GMP levels after brief flash illumination requires approximately twice as much time as the recovery of the membrane current. In contrast, the subsecond kinetics of the cyclic GMP response to light are faster than the kinetics of membrane current suppression. Both cyclic GMP and the membrane current show graded responses to a wide range of flash intensities; however, in a low Ca2+-Ringer's solution, dim flashes can trigger a decrease in cyclic GMP concentration with no corresponding decrease in membrane current. These results suggest that either other factors can regulate the membrane current, or that measurements of total cellular cyclic GMP do not accurately reflect dynamic changes in cyclic GMP concentration in the vicinity of the light-regulated channel. Changes in cyclic GMP concentration in the presence of background illumination exhibit adaptational behavior similar to that observed in a light-adapted photoresponse: acceleration in the response kinetics and a decrease in response amplitude. That this result is observed in rods depleted of internal Ca2+ suggests a Ca2+-independent mechanism by which background illumination can accelerate the cyclic GMP response.

Highlights

  • Illumination Alters the Response of Cyclic GMP 1).In contrast, within8 s after the onseotf the flash stimulus, to Flash Illumination-In thepresence of dimcontinuous cyclic GMP levels are not significantly different ( t test, p

  • We report that the response of cyclic GMP levels in isolated rod photoreceptors to flash illumination is graded with light intensity

  • Changes in free intracellular Ca2+that would normally occur during photoreceptor illumination (Ratto et al, 1988)are most likely reduced or eliminated in OSIS depleted of their intracellular Ca2+.As a result,the effects of illumination oncyclic GMP levels can be evaluated, while reducing the likelihood that changes in intracellular Ca2+might indirectly influence cyclic GMP metabolism

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Summary

Incontrasttothe consensus that has recently emerged

Scientific contribution No 1611 from the New Hampshire Agricultural Experiment Station. We employ suspensions of isolated, purified frog rod photoreceptors for direct comparisons of the biochemical and electrophysiological responses to flash illumination. This cellular preparation allows us to address three specific issues: 1)the relationship of changes in cyclic GMP concentration to themagnitude of membrane current during the onset of flash illumination;2) the time course of recovery of the response to a light flash; 3) the ability of cyclic GMP to exhibitlight-adapted behavior in the presence of background illumination. The rate of rhodopsin bleaching was quantitated asabove.Dark-adapted OSIS from another rod suspension were drawn into the suction pipette, and the flash intensity-response amplitude relation was determined using the diffuse illumination. Optical components for the light sources were purchased from Oriel and Edmund Scientific

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