Association of Serotonin, Dopamine, or Noradrenaline with an Actin‐Like Component in Pheochromocytoma (PC12) Cells

Abstract: A rat pheochromocytoma (PC12) cell line was used to examine the possibility that 5‐hydroxytryptamine (serotonin), 3,4‐dihydroxyphenylethylamine (dopamine), or noradrenaline may be associated with cytoplasmic actin, as was suggested by previous in vitro binding studies on an actin‐like protein from rat brain synaptosomes. When PC12 cells were incubated with [3H]serotonin, [3H]dopamine, or [3H]noradrenaline for 30 min at 37°C, approximately 2–4% of the radioactivity present in the cells was found to be associated with a high‐molecular‐weight (actin‐like) component in supernatant fractions. Evidence relating this monoamine binding component to actin filaments includes: (a) its strong absorption by myosin filaments at low ionic strength; (b) a decrease in its affinity for myosin in the presence of 1 mM ATP, which lowers the affinity of authentic actin for myosin; (c) displacement of bound [3H]serotonin from it by DNase I, which binds strongly to actin and which inhibits [3H]serotonin binding to actin in vitro; (d) an increase in its binding of each monoamine (by 25–40%) after PC12 cells were preincubated with 10 μM cytochalasin B (a drug that induces depolymerization of F‐actin). These findings suggest that serotonin, dopamine, or noradrenaline may associate with actin filaments in vivo.

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