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Vol. 9, Issue 8, 2305-2324, August 1998
q/11, G-Protein Receptor Kinase-2/3, and
-Arrestin-1/2
§
Departments of
*Surgery and
Observations in reconstituted systems and transfected cells
indicate that G-protein receptor kinases (GRKs) and Physiology, University
of California San Francisco, San Francisco, California 94143-0660
-arrestins mediate desensitization and endocytosis of G-protein-coupled
receptors. Little is known about receptor regulation in neurons.
Therefore, we examined the effects of the neurotransmitter substance P
(SP) on desensitization of the neurokinin-1 receptor (NK1-R) and on the
subcellular distribution of NK1-R, G
q/11, GRK-2 and -3, and -arrestin-1 and -2 in cultured myenteric neurons. NK1-R was coexpressed with immunoreactive Gq/11, GRK-2 and -3, and -arrestin-1 and -2 in a subpopulation of neurons. SP caused 1) rapid
NK1-R-mediated increase in [Ca2+]i, which
was transient and desensitized to repeated stimulation; 2)
internalization of the NK1-R into early endosomes containing SP; and 3)
rapid and transient redistribution of -arrestin-1 and -2 from the
cytosol to the plasma membrane, followed by a striking redistribution
of -arrestin-1 and -2 to endosomes containing the NK1-R and SP. In
SP-treated neurons Gq/11 remained at the plasma
membrane, and GRK-2 and -3 remained in centrally located and
superficial vesicles. Thus, SP induces desensitization and endocytosis
of the NK1-R in neurons that may be mediated by GRK-2 and -3 and
-arrestin-1 and -2. This regulation will determine whether
NK1-R-expressing neurons participate in functionally important reflexes.
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