The purpose of this core is to provide and maintain equipment, reagents, and expertise necessary to examine the localization of receptors for excitatory neurotransmitters at the cellular and subcellular level. Given the primary sequence information now available for many subtypes of excitatory transmitter receptors, straight-forward techniques are available to obtain highly selective labels for these molecules that can be applied in anatomical investigations. The emphasis of this facility will be on the development of agents and methods for immunocytochemical studies. Polyclonal antibodies that label specific receptors for excitatory transmitters will be generated using the anti-peptide antibody approach. Some anti-receptor antibodies are presently available through collaborators; these pertain to acetylcholine and glutamate receptor subtypes that were cloned several years ago. The applicants have successfully raised and purified antibodies against a newly described glutamate receptor (mGluR5) and are currently purifying antibodies we have raised against a recently cloned NMDA receptor (NR1). One of our mGluR5 antibodies appears to be highly specific for this receptor subtype. We have successfully applied this antibody for light microscopic immunocytochemical localization studies and are currently developing methods for immunoelectron microscopic application of this antibody. We have made arrangements to receive from Dr. Shigetada Nakanishi cDNA clones for various glutamate receptor subtypes which we will use to develop cell lines that express these receptors. These cell lines will provide an additional valuable means of confirming the receptor subtype specificity of the antibodies we generate. Image analysis hardware, as well as the equipment for light and electron microscopy are already available and are not requested for the core. The immunological probes and methods developed by this core will be a valuable asset for several of the projects in this research program.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Program Projects (P01)
Project #
3P01AG011355-05S1
Application #
6217021
Study Section
Project Start
1999-09-01
Project End
2000-08-14
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
5
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Washington University
Department
Type
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
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