Kondo H, Miller W H
Department of Ophthalmology and Visual Science, Yale Medical School, New Haven, CT 06510.
Proc Natl Acad Sci U S A. 1988 Feb;85(4):1322-6. doi: 10.1073/pnas.85.4.1322.
We compare the retinal rod photocurrent before and after introduction of an hydrolysis-resistant analog of GTP into the outer segment by the whole-cell patch technique. Others have shown that GTP bound to transducin leads to the hydrolysis of cyclic GMP, causing the response to light--a decrease in dark current. The hydrolysis-resistant GTP analog prolongs the response to a bright flash, which leads us to suggest that prolonged transducin activation by bright light desensitizes the rod by a prolonged decrease in dark current. Recovery from the response to a bright flash does occur after introduction of the analog; that recovery requires acceleration of cyclase activity rather than inhibition of phosphodiesterase. The analog mimics light adaptation by desensitizing the rod and speeding the recovery from a dim flash. The analog plus light or light adaptation prolongs the activities of transducin and phosphodiesterase (oligonucleate 5'-nucleotidohydrolase, EC 3.1.4.1) to mediate desensitization by reducing the dark current. Hence, this faster recovery from a dim flash would be by increased activity of guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] rather than by inhibited phosphodiesterase. Accelerated activity of guanylate cyclase may speed recovery by response truncation. We conclude that transducin, activated by photolyzed rhodopsin, may lead to increased activity of both phosphodiesterase and guanylate cyclase to mediate the desensitization and the faster recovery of the light-adapted response.
我们通过全细胞膜片钳技术,比较了将一种抗水解的GTP类似物引入视杆细胞外段前后的视网膜视杆细胞光电流。其他人已经表明,与转导蛋白结合的GTP会导致环鸟苷酸水解,从而引起对光的反应——暗电流减少。这种抗水解的GTP类似物延长了对强光闪光的反应,这使我们认为强光导致的转导蛋白长时间激活会通过暗电流的长时间减少使视杆细胞脱敏。在引入该类似物后,确实会从对强光闪光的反应中恢复;这种恢复需要环化酶活性的加速,而不是磷酸二酯酶的抑制。该类似物通过使视杆细胞脱敏并加速从弱光闪光中的恢复来模拟光适应。该类似物加光或光适应会延长转导蛋白和磷酸二酯酶(寡核苷酸5'-核苷酸水解酶,EC 3.1.4.1)的活性,通过降低暗电流来介导脱敏。因此,从弱光闪光中更快的恢复将是由于鸟苷酸环化酶[GTP焦磷酸裂解酶(环化),EC 4.6.1.2]活性的增加,而不是磷酸二酯酶的抑制。鸟苷酸环化酶活性的加速可能通过反应截断来加速恢复。我们得出结论,由光解视紫红质激活的转导蛋白可能导致磷酸二酯酶和鸟苷酸环化酶的活性增加,以介导脱敏和光适应反应的更快恢复。