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Study of neurotrophin-3 signaling in primary cultured neurons using multiplex stable isotope labeling with amino acids in cell culture

Study of neurotrophin-3 signaling in primary cultured neurons using multiplex stable isotope labeling with amino acids in cell culture
Study of neurotrophin-3 signaling in primary cultured neurons using multiplex stable isotope labeling with amino acids in cell culture
Conventional stable isotope labeling with amino acids in cell culture (SILAC) requires extensive
metabolic labeling of proteins, and therefore is difficult to apply to cells that do not divide or are
unstable in SILAC culture. Using two different sets of heavy amino acids for labeling allows for
straightforward SILAC quantitation using partially labeled cells because the two cell populations
are always equally labeled. Here we report the application of this labeling strategy to primary
cultured neurons. We demonstrated that protein quantitation was not compromised by incomplete
labeling of the neuronal proteins. We used this method to study neurotrophin-3 (NT-3) signaling
in primary cultured neurons. Surprisingly our results indicate TrkB signaling is a major
component of the signaling network induced by NT-3 in cortical neurons. In addition, involvement
of proteins such as VAMP2, Scamp1 and Scamp3 suggest NT-3 may lead to enhanced exocytosis
of synaptic vesicles.
1535-3893
2546-2554
Zhang, Guoan
fbb6a65f-dba6-43b8-aa9b-9a95f8c8a979
Deinhardt, Katrin
5f4fe23b-2317-499f-ba6d-e639a4885dc1
Chao, Moses V.
823c00e2-9f32-4a7b-a2c4-76e7c1fbf83d
Neubert, Thomas A.
bd2d3132-b85d-49fa-b879-488ac0e32ec7
Zhang, Guoan
fbb6a65f-dba6-43b8-aa9b-9a95f8c8a979
Deinhardt, Katrin
5f4fe23b-2317-499f-ba6d-e639a4885dc1
Chao, Moses V.
823c00e2-9f32-4a7b-a2c4-76e7c1fbf83d
Neubert, Thomas A.
bd2d3132-b85d-49fa-b879-488ac0e32ec7

Zhang, Guoan, Deinhardt, Katrin, Chao, Moses V. and Neubert, Thomas A. (2011) Study of neurotrophin-3 signaling in primary cultured neurons using multiplex stable isotope labeling with amino acids in cell culture. Journal of Proteome Research, 10 (5), 2546-2554. (doi:10.1021/pr200016n).

Record type: Article

Abstract

Conventional stable isotope labeling with amino acids in cell culture (SILAC) requires extensive
metabolic labeling of proteins, and therefore is difficult to apply to cells that do not divide or are
unstable in SILAC culture. Using two different sets of heavy amino acids for labeling allows for
straightforward SILAC quantitation using partially labeled cells because the two cell populations
are always equally labeled. Here we report the application of this labeling strategy to primary
cultured neurons. We demonstrated that protein quantitation was not compromised by incomplete
labeling of the neuronal proteins. We used this method to study neurotrophin-3 (NT-3) signaling
in primary cultured neurons. Surprisingly our results indicate TrkB signaling is a major
component of the signaling network induced by NT-3 in cortical neurons. In addition, involvement
of proteins such as VAMP2, Scamp1 and Scamp3 suggest NT-3 may lead to enhanced exocytosis
of synaptic vesicles.

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Published date: 2011
Organisations: Faculty of Natural and Environmental Sciences, Centre for Biological Sciences

Identifiers

Local EPrints ID: 345196
URI: http://eprints.soton.ac.uk/id/eprint/345196
ISSN: 1535-3893
PURE UUID: e49ff4be-0b8a-4923-93bd-195e65097170
ORCID for Katrin Deinhardt: ORCID iD orcid.org/0000-0002-6473-5298

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Date deposited: 13 Nov 2012 10:30
Last modified: 26 Nov 2019 01:36

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