Tracking transmitter-gated P2X cation channel activation in vitro and in vivo.
We present a noninvasive approach to track activation of ATP-gated P2X receptors and potentially other transmitter-gated cation channels that show calcium fluxes. We genetically engineered rat P2X receptors to carry calcium sensors near the channel pore and tested this as a reporter for P2X(2) receptor opening. The method has several advantages over previous attempts to image P2X channel activation by fluorescence resonance energy transfer (FRET): notably, it reports channel opening rather than a conformation change in the receptor protein. Our FRET-based imaging approach can be used as a general method to track, in real time, the location, regional expression variation, mobility and activation of transmitter-gated P2X channels in living neurons in vitro and in vivo. This approach should help to determine when, where and how different receptors are activated during physiological processes.
Richler
Esther
E
Department of Physiology, University of California Los Angeles, 10833 LeConte Avenue, Los Angeles, California 90095, USA.
Chaumont
Severine
S
Shigetomi
Eiji
E
Sagasti
Alvaro
A
Khakh
Baljit S
BS
D002118:Calcium / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D015640:Ion Channel Gating / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058469:Receptors, Purinergic P2X
D018048:Receptors, Purinergic P2; D058469:Receptors, Purinergic P2X; D002118:Calcium