Activation-dependent changes in receptor distribution and dendritic morphology in hippocampal neurons expressing P2X2-green fluorescent protein receptors.
ATP-gated P2X(2) receptors are widely expressed in neurons, but the cellular effects of receptor activation are unclear. We engineered functional green fluorescent protein (GFP)-tagged P2X(2) receptors and expressed them in embryonic hippocampal neurons, and report an approach to determining functional and total receptor pool sizes in living cells. ATP application to dendrites caused receptor redistribution and the formation of varicose hot spots of higher P2X(2)-GFP receptor density. Redistribution in dendrites was accompanied by an activation-dependent enhancement of the ATP-evoked current. Substate-specific mutant T18A P2X(2)-GFP receptors showed no redistribution or activation-dependent enhancement of the ATP-evoked current. Thus fluorescent P2X(2)-GFP receptors function normally, can be quantified, and reveal the dynamics of P2X(2) receptor distribution on the seconds time scale.
Khakh
B S
BS
Division of Biology, 156-29, California Institute of Technology, Pasadena, CA 91125, USA. bsk@mrc-lmb.cam.ac.uk [corrected]
Smith
W B
WB
Chiu
C S
CS
Ju
D
D
Davidson
N
N
Lester
H A
HA
D000255:Adenosine Triphosphate / Q000494:pharmacology; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D048429:Cell Size / Q000187:drug effects; D003712:Dendrites / Q000187:drug effects / Q000378:metabolism*; D004553:Electric Conductivity; D018698:Glutamic Acid / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology* / Q000196:embryology; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D055095:Optics and Photonics; D011554:Pseudopodia / Q000187:drug effects; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000378:metabolism*; D058476:Receptors, Purinergic P2X2; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012845:Sindbis Virus / Q000235:genetics; D014982:Xenopus laevis
D018048:Receptors, Purinergic P2; D058476:Receptors, Purinergic P2X2; D011993:Recombinant Fusion Proteins; D018698:Glutamic Acid; D000255:Adenosine Triphosphate