PMID- 17826844
OWN - NLM
STAT- MEDLINE
DCOM- 20080214
LR  - 20191003
IS  - 0165-0270 (Print)
IS  - 0165-0270 (Linking)
VI  - 166
IP  - 2
DP  - 2007 Nov 30
TI  - A rapid screening method for population-specific neuronal motogens, substrates 
      and associated signaling pathways.
PG  - 178-94
AB  - We developed and characterized an assay that allows for rapid examination of 
      migration of specific neuronal populations within a mixed population using the 
      Boyden chamber principle. Migration of cerebellar interneurons and granule cells 
      was examined using mice expressing enhanced green fluorescent protein (eGFP) 
      under the glutamate decarboxylase (GAD(65)) and growth-associated protein-43 
      (GAP43) promoters, respectively. Brain-derived neurotrophic factor (BDNF) was 
      used as the prototypic motogen for both populations. Fluorescent light-blocking 
      inserts (FluoroBlok) with different pore sizes and densities were compared in a 
      two-compartment assay. Immunodetection of polarity markers and nuclear staining 
      indicated that dendrites and somata are preferentially extended through the pores 
      in response to BDNF. Inserts coated with extracellular matrix (ECM) proteins were 
      used to examine interactions between BDNF and the ECM during migration. ECM 
      proteins alone stimulated migration when the lower side of the insert was coated, 
      however coating of both sides of the insert slowed migration when compared to 
      poly-D-lysine. Addition of a PI 3-kinase inhibitor to the lower compartment 
      blocked BDNF-stimulated migration of both populations while a Src inhibitor 
      reduced laminin-stimulated migration of interneurons, but not granule cells. We 
      also examined use of neurons cultured from GAD(65)-eGFP mice as a reporter system 
      for promoter activity. GAD(65)-eGFP mice may also be useful as a model for 
      promoter regulation and the potential confounding effects of eGFP induction by 
      the stimuli are also addressed. This assay allows for rapid analysis of motogens, 
      substrates and signaling pathways that regulate migration of selected neuronal 
      populations.
FAU - Hassoun, Amani T
AU  - Hassoun AT
AD  - Laboratory for Integrative Neuroscience/Section on Synaptic Pharmacology, 
      National Institute on Alcohol Abuse and Alcoholism, National Institutes of 
      Health, Bethesda, MD 20892, USA. hassouna@mail.nih.gov
FAU - Erdelyi, Ferenc
AU  - Erdelyi F
FAU - Szabo, Gabor
AU  - Szabo G
FAU - Davis, Margaret I
AU  - Davis MI
LA  - eng
GR  - Z99 AA999999/Intramural NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Intramural
DEP - 20070722
PL  - Netherlands
TA  - J Neurosci Methods
JT  - Journal of neuroscience methods
JID - 7905558
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Drug Combinations)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Fibronectins)
RN  - 0 (GAP-43 Protein)
RN  - 0 (Laminin)
RN  - 0 (Nerve Growth Factors)
RN  - 0 (Proteoglycans)
RN  - 119978-18-6 (matrigel)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 9007-34-5 (Collagen)
RN  - EC 4.1.1.15 (Glutamate Decarboxylase)
RN  - EC 4.1.1.15 (glutamate decarboxylase 2)
RN  - P658DCA9XD (neurotrophin 4)
SB  - IM
MH  - Age Factors
MH  - Animals
MH  - Animals, Newborn
MH  - Brain/*cytology/growth & development
MH  - Brain-Derived Neurotrophic Factor/pharmacology
MH  - Cell Migration Assays/methods
MH  - Cell Movement/drug effects/*genetics
MH  - Cells, Cultured
MH  - Collagen/pharmacology
MH  - Dose-Response Relationship, Drug
MH  - Drug Combinations
MH  - Enzyme Inhibitors/pharmacology
MH  - Fibronectins/metabolism
MH  - GAP-43 Protein/genetics/metabolism
MH  - Genetic Testing/*methods
MH  - Glutamate Decarboxylase/genetics/metabolism
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Laminin/pharmacology
MH  - Mice
MH  - Mice, Transgenic
MH  - Nerve Growth Factors/pharmacology
MH  - Neurons/*physiology
MH  - Proteoglycans/pharmacology
MH  - Signal Transduction/genetics/*physiology
PMC - PMC2443863
MID - NIHMS34503
EDAT- 2007/09/11 09:00
MHDA- 2008/02/15 09:00
PMCR- 2008/07/08
CRDT- 2007/09/11 09:00
PHST- 2007/04/02 00:00 [received]
PHST- 2007/07/02 00:00 [revised]
PHST- 2007/07/07 00:00 [accepted]
PHST- 2007/09/11 09:00 [pubmed]
PHST- 2008/02/15 09:00 [medline]
PHST- 2007/09/11 09:00 [entrez]
PHST- 2008/07/08 00:00 [pmc-release]
AID - S0165-0270(07)00363-9 [pii]
AID - 10.1016/j.jneumeth.2007.07.008 [doi]
PST - ppublish
SO  - J Neurosci Methods. 2007 Nov 30;166(2):178-94. doi: 
      10.1016/j.jneumeth.2007.07.008. Epub 2007 Jul 22.