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Journal of Neuroscience, Vol 13, 4281-4292, Copyright © 1993 by Society for Neuroscience
Signal transduction events mediated by the BDNF receptor gp 145trkB in primary hippocampal pyramidal cell culture
HN Marsh, WK Scholz, F Lamballe, R Klein, V Nanduri, M Barbacid and HC Palfrey
Department of Pharmacological and Physiological Sciences, University of Chicago, Illinois 60637.
The trkB gene encodes a tyrosine kinase receptor, gp145trkB, for brain-
derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4). To understand
the role of gp145trkB in the nervous system, we have investigated its
expression in embryonic rat hippocampal pyramidal cell cultures and
examined the effects of BDNF on signal transduction in the primary neurons.
The expression of trkB transcripts was established by PCR analysis and in
situ hybridization. In addition to gp145trkB, the pyramidal neuronal
cultures expressed transcripts specific for the NT-3 receptor gp145trkC,
but not for the high-affinity NGF receptor gp140trk or for p75LNGFR, a
low-affinity receptor for all known members of the NGF family of
neurotrophins including the gp145trkB ligands, BDNF and NT-4. The presence
of gp145trkB receptors in the primary neuronal cultures was confirmed by
immunocytochemical analysis in which > 90% of the cells stained with
affinity-purified polyclonal antibodies to gp145trkB. Immunoblots using
this antibody revealed a single approximately 140 kDa protein in both adult
hippocampus and pyramidal cultures. Addition of recombinant BDNF to these
cultures induced the tyrosine phosphorylation of gp145trkB, as determined
by antiphosphotyrosine staining of gp145trkB immunoprecipitates. Moreover,
BDNF treatment activated the microtubule-associated protein (MAP) kinases,
as determined by an increase in MAP2 phosphorylation in vitro. Both the 41
and 44 kDa forms of MAP kinase were activated by BDNF. BDNF also increased
c-fos expression in over 90% of the cells. These results indicate that
gp145trkB does not require p75LNGFR to form a functional receptor for BDNF
in hippocampal pyramidal neurons.
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