Recording single-channel activity of inositol trisphosphate receptors in intact cells with a microscope, not a patch clamp.
Optical single-channel recording is a novel tool for the study of individual Ca2+-permeable channels within intact cells under minimally perturbed physiological conditions. As applied to the functioning and spatial organization of IP3Rs, this approach complements our existing knowledge, which derives largely from reduced systems - such as reconstitution into lipid bilayers and patch clamping of IP3Rs on the membrane of excised nuclei - where the spatial arrangement and interactions among IP3Rs via CICR are disrupted. The ability to image the activity of single IP3R channels with millisecond resolution together with localization of their positions with a precision of a few tens of nanometers both raises several intriguing questions and holds promise of answers. In particular, what mechanism underlies the anchoring of puffs and blips to static locations; why do these Ca2+ release events appear to involve only a very small fraction of the IP3Rs within a cell; and how can we reconcile the relative immotility of functional IP3Rs with numerous studies reporting free diffusion of IP3R protein in the ER membrane?
Parker
Ian
I
Department of Neurobiology and Behavior, University of California, Irvine, Irvine, CA 92697, USA.
Smith
Ian F
IF
D000818:Animals; D002118:Calcium / Q000502:physiology; D015220:Calcium Channels / Q000502:physiology*; D020013:Calcium Signaling / Q000502:physiology; D006801:Humans; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000502:physiology*; D015640:Ion Channel Gating / Q000502:physiology; D008853:Microscopy / Q000379:methods; D018408:Patch-Clamp Techniques / Q000379:methods
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D002118:Calcium