期刊
NATURE METHODS
卷 5, 期 1, 页码 87-93出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH1144
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资金
- NIGMS NIH HHS [T32 GM065823] Funding Source: Medline
- NINDS NIH HHS [F31 NS064794] Funding Source: Medline
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [T32GM065823] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [F31NS064794] Funding Source: NIH RePORTER
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.
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