Functional Role Of Purinergic Receptor In Retinal Pigment Epithelial Cells

Objective Observe and understand the role of purinergic receptors in retinal pigment epithelial cell function.Method Application of FURA/2fluorescence lifetime imaging microscopy detect ion of ARPE-19cells, the calcium ion concentration. And by RT-PCR experimen t further identification of BK,SK and IK channels exist, and then the inside out, use the diaphragm experiment further identification of BK,SK and IK channels exist and the current density and open probability. Finally, through the use of a clathrin or caveolin protein (caveolin-1) for the media transport mechanism, che ck whether ATP through Ga Q reduced cell surface Kir4.1richness, and the use of Western blot protein imprinting analysis, analysis of caveolin and clathrin hea vy chain.Result The experimental results show that the application of ATP to increase the calcium concentration within the RPE cells by activating the purinergic receptor. The increase in intracellular calcium concentration able to open the calcium acti vated potassium channels. Many organizations in the SK channel in P2Y receptor and SK channel in the purine stimulation of P2Y receptor activation. The coupl ing of the P2Y receptor and SK channel activation protein of dependence Phoph olipase C activation and IP3production. IP3receptor manipulation Reserves due to local calcium release appears to be in SK stimulate. In this study, activation of the receptor can not only cause the increase in BK channel open probability, but also caused the increase in SK channel open probability. SK channel activati on leads to the outward currents to reduce membrane excitability. We also obtain ed a long time to cultivate the cause of inward rectifier potassium channel curre nt density decreases ATP mechanisms involved in endocytosis through a caveolin.Conclusion ATP has an important regulatory role both in calcium-activated pota ssium channels and Kir4.1channels. Purinergic activation increased the the open probability of SK channels as well as one of the BK channels, decreased the cu rrent density of Kir4.1through a mechanism involving caveolin-mediated endocyt osis.