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The Journal of Neuroscience, October 1, 2000, 20(19):7325-7333
NGF Signals through TrkA to Increase Clathrin at the Plasma
Membrane and Enhance Clathrin-Mediated Membrane Trafficking
Eric C.
Beattie1,
Charles L.
Howe4,
Andrew
Wilde2, 3, 5,
Frances M.
Brodsky2, 3, 5, and
William C.
Mobley6
Departments of 1 Physiology, 2 Immunology
and Microbiology, 3 Biopharmaceutical Sciences and
Pharmaceutical Chemistry, 4 Program in Neuroscience, and
5 G. W. Hooper Foundation, University of California at
San Francisco, San Francisco, California 94143, and
6 Departments of Neurology and Neurological Sciences,
Pediatrics, and the Program in Neuroscience, Stanford University,
Stanford, California 94305
Neurotrophin (NT) signals may be moved from axon terminals to
neuron cell bodies via signaling endosomes organelles in which NTs
continue to be bound to their activated receptors. Suggesting that
clathrin-coated membranes serve as one source of signaling endosomes,
in earlier studies we showed that nerve growth factor (NGF) treatment
increased clathrin at the plasma membrane and resulted in
colocalization of clathrin with TrkA, the receptor tyrosine kinase for
NGF. Strikingly, however, we also noted that most clathrin puncta at
the surface of NGF-treated cells did not colocalize with TrkA, raising
the possibility that NGF induces a general increase in clathrin-coated
membrane formation. To explore this possibility further, we examined
the distribution of clathrin in NGF- and BDNF-treated cells. NGF
signaling in PC12 cells robustly redistributed the adaptor protein AP2
and the clathrin heavy chain (CHC) to surface membranes. Using confocal
and epifluorescence microscopy, as well as biochemical assays, we
showed the redistribution of clathrin to be attributable to the
activation of TrkA. Significantly, NGF signaled through TrkA to induce
an increase in clathrin-mediated membrane trafficking, as revealed in
the increased endocytosis of transferrin. In that BDNF treatment
increased AP2 and clathrin at the surface membranes of hippocampal
neurons, these findings may represent a physiologically significant
response to NTs. We conclude that NT signaling increases
clathrin-coated membrane formation and clathrin-mediated membrane
trafficking and speculate that this effect contributes to their trophic
actions via the increased internalization of receptors and other
proteins that are present in clathrin-coated membranes.
Key words:
NGF; BDNF; neurotrophin; signaling; TrkA; clathrin; transferrin; endocytosis
Copyright © 2000 Society for Neuroscience 0270-6474/00/20197325-09$05.00/0
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