Accumulation and metabolism of choline and homocholine (cas 10238-59-2) by mouse brain subcellular fractions

Add time:07/26/2019    Source:sciencedirect.com

Minces of mouse forebrain were incubated in Krebs or a Krebs solution containing high K+ (32.7mM) and Li+, instead of Na+, for 30 min at 37°; subcellular fractions were prepared, and the levels of ACh in the S3 and P3 fractions were determined and compared for the two treatments. Incubation of minces in the Krebs solution with high K+ and Li+, instead of Na+, depleted the ACh content of the P3 fraction 70 per cent, without altering that of the S3 fraction with respect to incubation of minces in normal Krebs. Subsequent incubation of the depleted minces in normal Krebs containing [14C]choline (0.1 mM) and paraoxon (0.1μM) refilled the depleted P3 fraction with newly synthesized [14C]ACh, and the ratio of [14C]ACh to total ACh in this fraction (0.63) exceeded that of the S3 (0.33). Incubation of depleted minces in normal Krebs solution containing the choline analog [14C]homocholine (cas 10238-59-2) (0.1 mM) and paraoxon (0.1μM) also refilled the depleted P3 fraction with newly synthesized [14C]acetylhomocholine, and the ratio of [14C]acetylhomocholine to ACh in this fraction (7.26) exceeded that of the S3 (0.44). Refilling of the depleted P3 fraction was due to an increase in the accumulation of precursor ([14C]choline 84 per cent and [14C]homocholine, 76 per cent) which occurred independently of the S3. Incubation of depleted minces with either extracellular [14C]ACh or [14C]acetylhomocholine did not refill the depleted P3 fraction with these compounds. These results suggest that ACh, lost from the crude vesicular fraction, can be replaced with newly synthesized ACh independently of the cytoplasm.

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