Axonal alterations in demyelinating diseases

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DESCRIPTION (provided by applicant): We hypothesize that in normal myelinated PNS axons, the combination of Kv1.1, Kv1.2, KCNQ2, and KCNQ3 is necessary for repolarization, and that the misexpression of Kv3.1 b has a deleterious effect on axonal conduction of de/remyelinated axons. In unpublished work, we have found the alphas isoform of Na,K-ATPase is the only one that is clearly localized to axons that it is (surprisingly) excluded from nodes, and appears to be focally diminished by demyelination. Thus, we also hypothesize that the misexpression of alphas may contribute to depolarization of de/remyelinated axons. The proposed experiments build on these findings, with the central theme of illuminating how K+ homeostasis works in normal and de/remyelinated axons. Aim #1: Do Kv3.1b channels contribute to conduction failure in demyelinating diseases? We will investigate whether KvS.lb is the 4-AP-sensitive channel by comparing the effects of DTX-I and 4- AP on axonal conduction in TremblerJ mice on a KvS.lb -null (Kcncl-/-) versus Kcnc1+/+ background. Aim #2: What is the role of KCNQ2 in myelinated axons? Because Kcnq2-null mice die at birth, before myelination and the formation of nodes, we will generate a conditional Kcnq2-null mouse and analyze the structure and function of myelinated axons, including the expression of KCNQ3. Aim #3: What Na,K-ATPase isoforms are expressed by myelinated axons and demyelinated axons? we will localize alpha1-3 by immunoelectron microscopy in CNS and PNS myelinated axons, along with their beta subunits.