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Study of Distribution and Transport Events of the GluR1 AMPA Receptor: Combination of Genetically Modified Receptors and Multi-Resolution Microscopy

Published online by Cambridge University Press:  02 July 2020

A. Stelljes ,
E.A. Bushong ,
M.E. Martone ,
P.W. Wiseman ,
K.L. Hood ,
M. Mayford  and
M.H. Ellisman
A. Stelljes
Affiliation:
National Center for Microscopy and Imaging Research and Department of Neurosciences
E.A. Bushong
Affiliation:
Biomedical Sciences Program
M.E. Martone
Affiliation:
National Center for Microscopy and Imaging Research and Department of Neurosciences
P.W. Wiseman
Affiliation:
Graduate Program in Neurosciences
K.L. Hood
Affiliation:
Department of Chemistry and Biochemistry, Univ. of California, San Diego, La Jolla, CA, 92093, USA
M. Mayford
Affiliation:
Cell Biology, The Scripps Research Institute, La Jolla, CA, 92037, USA
M.H. Ellisman
Affiliation:
National Center for Microscopy and Imaging Research and Department of Neurosciences
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Abstract

The formation of memory and the process of learning are believed to be regulated, at least in part, by the expression, distribution, and redistribution of glutamate receptors. The expression of these receptors at synaptic sites has a major impact on the strength of synaptic connections, and the AMPA receptor subunit GluR1 appears to play a key role within this system. Increasing evidence suggests that previously silent synapses become activated through rapid AMPA receptor insertion upon appropriate stimulation, and thus the trafficking of this receptor subunit from cellular stores to the synapse is of prime interest.

We are using a transgenic mouse expressing a GFP-tagged form of GluR1 (GluR1-GFP) in order to study the dynamic changes in GluR1 expression and distribution occurring during brain development and following induction of long-term potentiation (LTP). The fusion protein is transcribed under the control of the CaMKIIα promoter, which restricts the localization to forebrain neurons and is coupled to a tetrepressible system, thus allowing for the control of transcription with doxycycline. The anatomical distribution of GluR1-GFP on the light level is consistent with that of wild-type GluR1 (Fig. 1).

We are combining molecular biology with quantitative 3D image analysis on the light and electron microscopic level. The highly fluorescent polar tracer Alexa Fluor 568 (Molecular Probes Inc.) is being used to fill GluR1-GFP expressing pyramidal neurons in tissue slices fixed with aldehydes

Type
Bridging the Gap Between Structural and Molecular Biology (Organized by B. Herman)
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 40 - 41
Copyright
Copyright © Microscopy Society of America 2001

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References

references

1.Lu, W.-Y.et al., Neuron 29(2001)243.CrossRefGoogle ScholarPubMed
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4. This research was supported by grants from NIH NCRR RR04050 to Mark H. Ellisman and NIH RO1MH57368-04 to Mark MayfordGoogle Scholar