Transport of choline in rat brain slices.
Enzyme-modified amperometric microsensors have been utilized in the investigation of acetylcholine and choline diffusion in solution and choline uptake and diffusion in rat brains. A small amount of the substance of interest was introduced by pressure injection and transport to the sensor was monitored. The apparent diffusion coefficients for acetylcholine and choline in agarose gel perfused with physiological solutions were determined to be 5.2 +/- 0.7 x 10(-6) cm2/s and 6.1 +/- 0.8 x 10(-6) cm2/s, respectively. Choline transport was monitored in two brain regions: the caudate and anterior hypothalamus. The transport time of choline in the caudate was concentration dependent, but was unaffected by the presence of a competitive, high-affinity uptake inhibitor, hemicholinium-3. The apparent diffusion coefficient (D) and uptake rate (k) for choline in the caudate and anterior hypothalamus were calculated using a model for point source diffusion coupled with first-order uptake kinetics. The effect of the sensors' response time on the measurements was removed by deconvolution. The D and k were 1.8 +/- 0.1 x 10(-6) cm2/s and 2.0 +/- 0.1 x 10(-2) s-1 in the caudate and 1.9 +/- 0.1 x 10(-6) cm2/s and 3.2 +/- 0.6 x 10(-2) s-1 in the anterior hypothalamus. The reduced diffusion coefficient determined in brain tissue compared to agar gel is consistent with the increased tortuosity of the brain microenvironment. A substance in brain tissue, presumably acetylcholinesterase, prevents the use of differential measurements of acetylcholine because choline sensors became sensitive to acetylcholine.
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Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290, USA.
Wightman
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RM
D000109:Acetylcholine / Q000493:pharmacokinetics; D000818:Animals; D016477:Artifacts; D001667:Binding, Competitive / Q000502:physiology; D015374:Biosensing Techniques; D002421:Caudate Nucleus / Q000378:metabolism*; D002794:Choline / Q000493:pharmacokinetics*; D018678:Cholinergic Agents / Q000493:pharmacokinetics; D002799:Cholinergic Fibers / Q000378:metabolism; D004058:Diffusion; D005782:Gels; D006426:Hemicholinium 3 / Q000493:pharmacokinetics; D007032:Hypothalamus, Anterior / Q000378:metabolism*; D008297:Male; D009924:Organ Culture Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D012685:Sepharose
D018678:Cholinergic Agents; D005782:Gels; D006426:Hemicholinium 3; D012685:Sepharose; D002794:Choline; D000109:Acetylcholine