Project: Research project

Project Details


The specific aim is to culture dissociated neurons from the locus
coeruleus (LC), substantia nigra (SN) and dorsal raphe (DR) and study of
the action of cocaine on single isolated neurons. With isolated cells the
direct action of cocaine on membrane properties and exogenously applied
transmitters can be studied quantitatively since diffusion barriers and
endogenous transmitters will be limited. Agonists can be applied in known
concentration close to the cell. The effect that cocaine has on the
kinetics of ligand activated conductances will be studied to determine if
cocaine changes the affinity of the receptor for the transmitter. In
addition, the use of cultured cells permits whole cell recording with
patch electrodes such that more control of the intracellular ion content
and second messengers is gained. The long-term goal is to use the
strengths of each; the brain slice and isolated cell preparations to study
action of drugs of abuse at the cellular level. In the work proposed
here, the acute and chronic actions of cocaine will be determined in cells
from relatively homogenous nuclei (LC, SN and DR). The knowledge gained
in these simple nuclei will be applied to the action(s) of cocaine in more
complex areas such as the ventral tegmental area, nucleus accumbens and
prefrontal cortex. Studies from cells taken from animals which have self-
administered cocaine may lead to an understanding of some of the changes
which cause or result from chronic cocaine use. Whole cell recordings
will also eventually be used to study the changes which occur with other
drugs of abuse such as opioids. Such recordings from LC neurons with
patch electrodes are vital to determine the nature of the reduction in the
opioid 'spare-receptor' population following chronic opioid treatment. This award will afford the PI the time to learn and become competent with
the tissue culture and whole cell recording methods. The proposed
experiments are a natural progression of those which are in progress in
the slice preparations. The knowledge and experience gained by the work
carried out on LC and DR neurons in the slice over the past 8 years will
be directly applicable to work on isolated cells. This coupled with the
expertise of members of the Vollum Institute in recording from cultured
neurons provide an optimistic setting from which the PI can learn and
apply this knowledge to the understanding of cellular mechanisms of drug
Effective start/end date2/1/901/31/95


  • National Institutes of Health


  • Medicine(all)


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