In PC12 cells, it has been previously reported that nerve growth factor stimulates neuropeptide Y (NPY) gene expression. In the current study we examined the signalling pathways involved in this effect by transiently expressing in PC12 cells the receptor (TrkB) for the related neurotrophin, brain-derived neurotrophic factor (BDNF). BDNF caused a 3-fold induction of luciferase expression from a transiently co-transfected plasmid possessing the firefly luciferase gene under the control of the NPY promoter. This effect of BDNF was completely blocked by either a Y484F mutation in TrkB (which blocks high-affinity Shc binding to TrkB) or by a Y785F substitution [which blocks the binding, phosphorylation and activation of phospholipase Cγ (PLCγ)]. Activation of the NPY promoter by neurotrophin-3 in PC12 cells overexpressing TrkC was also completely blocked by a naturally occurring kinase insert which prevents the high-affinity binding of Shc and PLCγ. NPY promoter activation by BDNF was blocked by PD98059, suggesting a role for mitogen-activated protein kinase (MAP kinase). Stimulation of NPY gene expression by PMA, but not by BDNF, was blocked by Ro-31-8220, a protein kinase C inhibitor, excluding a role for this serine/threonine protein kinase in the effect of BDNF. In addition, BDNF did not cause an elevation in cytosolic Ca2+ concentration. Taken together, our results suggest that stimulation of the NPY promoter by BDNF requires the simultaneous activation of two distinct pathways; one involves Shc and MAP kinase, and the other appears to be PLCγ-independent but requires an intact tyrosine-785 on TrkB and so may involve an effector of TrkB signalling that remains to be identified.
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Research Article|
August 01 1998
Stimulation of neuropeptide Y gene expression by brain-derived neurotrophic factor requires both the phospholipase Cγ and Shc binding sites on its receptor, TrkB
Alan G. WILLIAMS;
Alan G. WILLIAMS
*Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, U.K.
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Andrew C. HARGREAVES;
Andrew C. HARGREAVES
†Department of Anatomy, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, U.K.
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Frank J. GUNN-MOORE;
Frank J. GUNN-MOORE
*Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, U.K.
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Jeremy M. TAVARÉ
Jeremy M. TAVARÉ
1
*Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, U.K.
1To whom correspondence should be addressed (e-mail j.tavare@bristol.ac.uk).
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Publisher: Portland Press Ltd
Received:
February 26 1998
Revision Received:
April 27 1998
Accepted:
May 12 1998
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 333 (3): 505–509.
Article history
Received:
February 26 1998
Revision Received:
April 27 1998
Accepted:
May 12 1998
Citation
Alan G. WILLIAMS, Andrew C. HARGREAVES, Frank J. GUNN-MOORE, Jeremy M. TAVARÉ; Stimulation of neuropeptide Y gene expression by brain-derived neurotrophic factor requires both the phospholipase Cγ and Shc binding sites on its receptor, TrkB. Biochem J 1 August 1998; 333 (3): 505–509. doi: https://doi.org/10.1042/bj3330505
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