Regulation of an Aplysia bag-cell neuron cation channel by closely associated protein kinase A and a protein phosphatase.

Ion channel regulation by closely associated kinases or phosphatases has emerged as a key mechanism for orchestrating neuromodulation. An exemplary case is the nonselective cation channel that drives the afterdischarge in Aplysia bag cell neurons. Initial studies showed that this channel is modulated by both a closely associated PKC and a serine/threonine protein phosphatase (PP). In excised, inside-out patches, the addition of ATP (a phosphate source) increases open probability (P(O)) through PKC, and this is reversed by the PP. Previous work also reported that, in certain cases, ATP can decrease cation channel P(O). The present study characterizes and provides a mechanism for this decreased P(O) ATP response. The kinetic change for channels inhibited by ATP was identical to the previously reported effect of exogenously applied protein kinase A (PKA) (i.e., a lengthening of the third closed-state time constant). The decreased P(O) ATP response was blocked by the PKA inhibitor peptide PKA(6-22), and its reversal was prevented by the PP inhibitor microcystin-LR. Furthermore, PKA(6-22) did not alter the increased P(O) ATP response. This suggests that both PKA and a PP are closely associated with these cation channels, but PKA and PKC are not simultaneously targeted. After an afterdischarge, the bag cell neurons are refractory and fail to respond to subsequent stimulation. The association of PKA with the cation channel may contribute to this decrease in excitability. Altering the constituents of a regulatory complex, such as exchanging PKA for PKC, may represent a general mechanism to precisely control ion channel function and excitability.