Shulman L M, Fox D A
College of Optometry, University of Houston, TX 77204-6052, USA.
Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):8034-9. doi: 10.1073/pnas.93.15.8034.
The rat retina contains dopaminergic interplexiform cells that send processes to the outer plexiform layer where dopamine is released in a light-dependent manner. We report herein that physiologically relevant concentrations of dopamine inhibited ouabain-sensitive photoreceptor oxygen consumption in dark- and light-adapted rat retinas and inhibited Na+,K+-ATPase specific activity (EC 3.6.1.37) in a rat rod outer-inner segment preparation. Experiments with the selective D1 agonist fenoldopam or D2 agonist quinpirole and experiments with dopamine plus either the D1 antagonist SCH23390 or D2/D4 antagonist clozapine showed that the inhibition of oxygen consumption and enzyme activity were mediated by D2/D4-like receptors. The amphetamine-induced release of dopamine, monitored by the inhibition of oxygen consumption, was blocked by L-2-amino-4-phosphonobutyric acid and kynurenic acid. Pharmacological and biochemical experiments determined that the IC50 values of ouabain for the alpha1-low and alpha3-high ouabain affinity isozymes of photoreceptor Na+,K+-ATPase were approximately 10(-5) and approximately 10(-7) M, respectively, and that the D2/D4-like mediated inhibition of Na+,K+-ATPase was exclusively selective for the alpha3 isozyme. The dopamine-mediated inhibition of alpha3 first occurred at 5 nM, was maximal at 100 microM (-47%), had an IC50 value of 382 +/- 23 nM, and exhibited negative cooperativity (Hill coefficient, 0.27). Prior homogenization of the rod outer-inner segment completely prevented the long-lasting inhibition, suggesting that the effect was coupled to a second messenger. Although the physiological significance of our findings to photoreceptor function is unknown, we hypothesize that these results may have relevance for the temporal tuning properties of rods.
大鼠视网膜含有多巴胺能双极细胞,其轴突伸向外网状层,在该层多巴胺以光依赖的方式释放。我们在此报告,生理相关浓度的多巴胺抑制暗适应和光适应大鼠视网膜中哇巴因敏感的光感受器氧消耗,并抑制大鼠视杆细胞外段 - 内段制剂中的Na +,K + -ATP酶比活性(EC 3.6.1.37)。用选择性D1激动剂非诺多泮或D2激动剂喹吡罗进行的实验,以及用多巴胺加D1拮抗剂SCH23390或D2 / D4拮抗剂氯氮平进行的实验表明,氧消耗和酶活性的抑制是由类D2 / D4受体介导的。通过抑制氧消耗监测到的苯丙胺诱导的多巴胺释放被L-2-氨基-4-膦酰丁酸和犬尿烯酸阻断。药理学和生化实验确定,哇巴因对光感受器Na +,K + -ATP酶的α1低亲和力和α3高亲和力同工酶的IC50值分别约为10(-5)和约10(-7)M,并且类D2 / D4介导的Na +,K + -ATP酶抑制仅对α3同工酶具有选择性。多巴胺介导的α3抑制首先在5 nM时出现,在100μM时最大(-47%),IC50值为382±23 nM,并表现出负协同性(希尔系数,0.27)。视杆细胞外段 - 内段的预先匀浆完全阻止了这种持久的抑制,表明这种作用与第二信使有关。虽然我们的发现对光感受器功能的生理学意义尚不清楚,但我们推测这些结果可能与视杆细胞的时间调谐特性有关。