PMID- 16837782
OWN - NLM
STAT- MEDLINE
DCOM- 20060919
LR  - 20111117
IS  - 1424-862X (Print)
IS  - 1424-862X (Linking)
VI  - 15
IP  - 1
DP  - 2006-2007
TI  - Cyclic-AMP response element-based signaling assays for characterization of Trk 
      family tyrosine kinases modulators.
PG  - 26-39
AB  - Neurotrophins (NTs) induce gene transcription by binding their high-affinity 
      tropomyosin-related kinase (Trk) receptors and initiating intracellular signal 
      transduction cascades. In particular, activation of the cyclic AMP response 
      element (CRE) in the promoters of target genes serves as surrogate markers for 
      Trk receptor activation as demonstrated in both in vivo and in vitro systems. We 
      used a HEK293 cell line stably expressing a CRE-luciferase reporter gene to 
      develop an assay for monitoring Trk activation in response to their cognate 
      ligands. Using TrkB, we showed that the assay was sensitive to physiological 
      concentrations of brain-derived neurotrophic factor (BDNF) and that the signal 
      was sufficiently robust to be suitable for implementation in high-throughput 
      format. Further characterization of the TrkB expressing stable cell lines showed 
      high-affinity binding for BDNF, a high density of receptor expression, and 
      supported BDNF-mediated phosphorylation signaling. Consistent with this, 
      inhibitors of phosphatidylinositol 3-kinase and the phospholipase C-gamma 
      pathways led to reduction of BDNF-mediated luciferase responses. In contrast, 
      inhibitors of mitogen-activated protein kinase pathways further potentiated BDNF 
      responses. This assay was NT-Trk receptor pair-selective and shown to be further 
      applicable to other Trk family members. This assay may be useful in screening 
      compound libraries to identify Trk agonists, which may be applied towards 
      discriminating between the activities of the different Trk receptor family 
      members and the development of pharmacological drugs.
FAU - Zhang, Jie
AU  - Zhang J
AD  - Neuroscience Discovery Research, Wyeth Research, Princeton, NJ 08543-8000, USA.
FAU - Chen, Diana
AU  - Chen D
FAU - Gong, Xiaohai
AU  - Gong X
FAU - Ling, Huaiping
AU  - Ling H
FAU - Zhang, Guoming
AU  - Zhang G
FAU - Wood, Andrew
AU  - Wood A
FAU - Heinrich, Julia
AU  - Heinrich J
FAU - Cho, Seongeun
AU  - Cho S
LA  - eng
PT  - Comparative Study
PT  - Journal Article
DEP - 20060706
PL  - Switzerland
TA  - Neurosignals
JT  - Neuro-Signals
JID - 101134359
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Carbazoles)
RN  - 0 (Cyclic AMP Response Element-Binding Protein)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Indole Alkaloids)
RN  - 97161-97-2 (staurosporine aglycone)
RN  - EC 1.13.12.- (Luciferases)
RN  - EC 2.7.10.1 (Receptor, trkB)
SB  - IM
MH  - Blotting, Western/methods
MH  - Brain-Derived Neurotrophic Factor/pharmacology
MH  - Carbazoles/pharmacology
MH  - Cell Line
MH  - Cyclic AMP Response Element-Binding Protein/*metabolism
MH  - Dose-Response Relationship, Drug
MH  - Drug Interactions
MH  - Enzyme Activation/drug effects
MH  - Enzyme Inhibitors/pharmacology
MH  - Gene Expression/drug effects
MH  - Humans
MH  - Indole Alkaloids
MH  - Luciferases/metabolism
MH  - Protein Binding/drug effects
MH  - Radioligand Assay/methods
MH  - Receptor, trkB/*metabolism
MH  - Signal Transduction/drug effects/*physiology
MH  - Time Factors
MH  - Transfection/methods
EDAT- 2006/07/14 09:00
MHDA- 2006/09/20 09:00
CRDT- 2006/07/14 09:00
PHST- 2006/04/04 00:00 [received]
PHST- 2006/06/01 00:00 [accepted]
PHST- 2006/07/14 09:00 [pubmed]
PHST- 2006/09/20 09:00 [medline]
PHST- 2006/07/14 09:00 [entrez]
AID - NSG2007015001026 [pii]
AID - 10.1159/000094385 [doi]
PST - ppublish
SO  - Neurosignals. 2006-2007;15(1):26-39. doi: 10.1159/000094385. Epub 2006 Jul 6.