The timing of endocytosis after activation of a G-protein-coupled receptor in a sensory neuron.
Endocytosis is a fundamental cellular event in membrane retrieval after exocytosis and in the regulation of receptor-mediated signal transduction. In contrast to the well-studied depolarization-induced membrane recycling, little is known about the kinetics of ligand-induced endocytosis of G-protein-coupled receptors in neurons. Here we investigated the kinetics of ligand-receptor binding-induced endocytosis in rat sensory neurons using a membrane capacitance assay. The time constant of ADP-induced endocytosis of P2Y-receptors was determined as 1.7 s. The ADP-induced endocytosis was blocked by antagonists against P2Y, phosphorylation, and clathrin. However, block of dynamin was without effect. The ADP-induced endocytosis was confirmed independently by a single vesicle image technique using a styryl FM2-10. Finally, the receptors were internalized in response to ADP, as determined by GFP-labeled P2Y. We conclude that ligand-receptor binding leads to rapid endocytosis in the cytoplasm of rat dorsal root ganglion neurons.
Wang
Lie-Cheng
LC
Institute of Molecular Medicine, Peking University, Beijing, China.
Xiong
Wei
W
Zheng
Jing
J
Zhou
Yang
Y
Zheng
Hui
H
Zhang
Chen
C
Zheng
Liang-Hong
LH
Zhu
Xue-Liang
XL
Xiong
Zhi-Qi
ZQ
Wang
Lu-Yang
LY
Cheng
He-Ping
HP
Zhou
Zhuan
Z
D000200:Action Potentials / Q000187:drug effects / Q000502:physiology*; D000244:Adenosine Diphosphate / Q000008:administration & dosage*; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology*; D008969:Molecular Sequence Data; D009475:Neurons, Afferent / Q000187:drug effects / Q000502:physiology; D020671:Posterior Horn Cells / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058472:Receptors, Purinergic P2Y1; D013997:Time Factors
C546382:P2RY1 protein, human; C546384:P2ry1 protein, rat; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D000244:Adenosine Diphosphate