Distinction among neuronal subtypes of voltage-activated sodium channels by mu-conotoxin PIIIA

The functional properties of most sodium channels are too similar to permit identification of specific sodium channel types underlying macroscopic cur rent. Such discrimination would be particularly advantageous in the nervous system in which different sodium channel family isoforms are coexpressed i n the same cell. To test whether members of the mu-conotoxin family can dis criminate among known neuronal sodium channel types, we examined six toxins for their ability to block different types of heterologously expressed sod ium channels. PIIIA mu-conotoxin blocked rat brain type II/IIA (rBII/IIA) a nd skeletal muscle sodium current at concentrations that resulted in only s light inhibition of rat peripheral nerve (rPN1) sodium current. Recordings from variant lines of PC12 cells, which selectively express either rBII/IIA or rPN1 channel subtypes, verified that the differential block by PIIIA al so applied to native sodium current. The sensitivity to block by PIIIA toxi n was then used to discriminate between rBII/IIA and rPN1 sodium currents i n NGF-treated PC12 cells in which both mRNAs are induced. During the first 24 hr of NGF-treatment, PN1 sodium channels accounted for over 90% of the s odium current. However, over the ensuing 48 hr period, a sharp rise in the proportion of rBII/IIA sodium current occurred, confirming the idea, based on previous mRNA measurements, that two distinct sodium channel types appea r sequentially during neuronal differentiation of PC12 cells.