TY - CHAP
T1 - ErbB membrane tyrosine kinase receptors
T2 - Analyzing migration in a highly complex signaling system
AU - Brossier, Nicole M.
AU - Byer, Stephanie J.
AU - Peavler, Lafe T.
AU - Carroll, Steven L.
N1 - Funding Information:
The development of the experimental protocols described in this chapter was supported by grants from the National Institute of Neurological Diseases and Stroke (R01 NS048353 to S.L.C.; F30 NS063626 to N.M.B.), the National Cancer Institute (R01 CA122804 to S.L.C.; R01 CA134773 to Kevin A. Roth and S.L.C.), and the Department of Defense (X81XWH-09-1-0086 to S.L.C.). L.T.P. was supported in part by a grant to the University of Alabama at Birmingham from the Howard Hughes Medical Institute through the Med into Grad Initiative. We thank the Alabama Neuroscience Blueprint Core Center (P30 NS57098) and the UAB Neuroscience Core Center (P30 NS47466) for technical assistance with studies from our laboratory that are described in this chapter. The content is solely the responsibility of the authors and does not necessarily represent the offi cial views of the National Institutes of Health or the Department of Defense.
Funding Information:
The development of the experimental protocols described in this chapter was supported by grants from the National Institute of Neurological Diseases and Stroke (R01 NS048353 to S.L.C.; F30 NS063626 to N.M.B.), the National Cancer Institute (R01 CA122804 to S.L.C.; R01 CA134773 to Kevin A. Roth and S.L.C.), and the Department of Defense (X81XWH-09-1-0086 to S.L.C.). L.T.P. was supported in part by a grant to the University of Alabama at Birmingham from the Howard Hughes Medical Institute through the Med into Grad Initiative. We thank the Alabama Neuroscience Blueprint Core Center (P30 NS57098) and the UAB Neuroscience Core Center (P30 NS47466) for technical assistance with studies from our laboratory that are described in this chapter. The content is solely the responsibility of the authors and does not necessarily represent the of fi cial views of the National Institutes of Health or the Department of Defense.
Publisher Copyright:
© Springer Science+Business Media, LLC 2012.
PY - 2012
Y1 - 2012
N2 - The erbB membrane tyrosine kinases (EGF receptor, erbB2, erbB3, and erbB4) are a family of structurally similar transmembrane proteins that act as receptors for the EGF and neuregulin (NRG) families of growth factors. ErbB receptors and their ligands are widely expressed by neurons and glia throughout the peripheral and central nervous system, where they promote proliferation, survival, migration, differentiation, and other effects. Precisely which effects are induced by erbB activation depends on a variety offactors. These factors include which erbB receptors are recruited to the signaling complex, whether key non-erbB intramembranous proteins (e.g., mucin 1, CD44, integrins) are present and interact with the erbB kinases and the lineage and developmental state of the cell. Different erbB ligands acting through the same receptors often also elicit distinct or even diametrically opposed effects. In this chapter, we review key aspects of the complexity intrinsic to signaling by erbB kinases and their ligands in the nervous system. We then relate this knowledge to the rational design of experiments examining erbB signaling in migration, a common response induced by erbB receptor activation. The proper performance of Boyden chamber assays is considered, together with the procedures used to identify appropriate migration substrates and to optimize key experimental parameters (cell number, migration time, comparisons of the effects of different erbB ligands, and establishing optimal concentrations of these growth factors). The use of pharmacologic inhibitors and RNA interference to establish whether specific erbB kinases are required for the migration of nervous system cells is also described.
AB - The erbB membrane tyrosine kinases (EGF receptor, erbB2, erbB3, and erbB4) are a family of structurally similar transmembrane proteins that act as receptors for the EGF and neuregulin (NRG) families of growth factors. ErbB receptors and their ligands are widely expressed by neurons and glia throughout the peripheral and central nervous system, where they promote proliferation, survival, migration, differentiation, and other effects. Precisely which effects are induced by erbB activation depends on a variety offactors. These factors include which erbB receptors are recruited to the signaling complex, whether key non-erbB intramembranous proteins (e.g., mucin 1, CD44, integrins) are present and interact with the erbB kinases and the lineage and developmental state of the cell. Different erbB ligands acting through the same receptors often also elicit distinct or even diametrically opposed effects. In this chapter, we review key aspects of the complexity intrinsic to signaling by erbB kinases and their ligands in the nervous system. We then relate this knowledge to the rational design of experiments examining erbB signaling in migration, a common response induced by erbB receptor activation. The proper performance of Boyden chamber assays is considered, together with the procedures used to identify appropriate migration substrates and to optimize key experimental parameters (cell number, migration time, comparisons of the effects of different erbB ligands, and establishing optimal concentrations of these growth factors). The use of pharmacologic inhibitors and RNA interference to establish whether specific erbB kinases are required for the migration of nervous system cells is also described.
KW - Boyden chamber
KW - Epidermal growth factor
KW - Membrane tyrosine kinase receptors
KW - Neuregulin
KW - Pharmacologic inhibitors
KW - RNA interference
UR - http://www.scopus.com/inward/record.url?scp=84979291919&partnerID=8YFLogxK
U2 - 10.1007/978-1-61779-824-5_7
DO - 10.1007/978-1-61779-824-5_7
M3 - Chapter
AN - SCOPUS:84979291919
T3 - Neuromethods
SP - 105
EP - 131
BT - Neuromethods
PB - Humana Press Inc.
ER -