Both cocktail solutions restored the elevated global [Ca2+]i responses and restored the NFATc1 nuclear translocation induced by high-K+ stimulation (Figure 9E, n = 12; Figure S1C, n = 6). Such data are summarized in Figures 9G and 9H (for statistics, see Supplemental Information). Taken together, our data are best explained by such local Ca2+i signals occurring

in microdomains containing AKAP79/150-orchestrated Ca2+-binding molecules, such as CaN and CaM, and L channels, which function as the activity reporter that links neuronal activity with NFAT-mediated Paclitaxel supplier transcriptional regulation (see Discussion). IM amplitudes and the expression level of KCNQ2 and KCNQ3 transcripts were also assayed under these same conditions. Consistent with the EGFP-NFATc1 translocation results, there was no enhancement of IM amplitudes in WT SCG neurons that had been stimulated with zero Ca2+-added (0.71 ± 0.09 pA/pF, n = 10), or nifedipine-added 50 K+ solutions (0.81 ± 0.05 pA/pF, n = 9), compared with neurons stimulated under regular Ringer’s solution (0.87 ± 0.05 pA/pF, n = 18) ( Figures

10A and 10B). qPCR was also performed from WT SCG neurons 7 hr after perfusion of regular Ringer’s, 50 K+, 50 K+ with CsA, or 50 K+ with nifedipine IWR 1 solutions for 15 min. We detected significant increases in the amount of both KCNQ2 and KCNQ3 mRNA in neurons depolarized by 50 K+ (3.2 ± 0.8 and 3.9 ± 0.7, n = 4; p < 0.01). This elevated Rolziracetam expression of KCNQ2 and KCNQ3 mRNA was suppressed when CsA (1.05 ± 0.20 and 1.41 ± 0.15, n = 4) or nifedipine (1.25 ± 0.28 and 1.61 ± 0.50, n = 4) was present during the stimulation ( Figure 10C). Thus, removal of external Ca2+, blockade of CaN, or addition of nifedipine during stimulation eliminates NFATc1 nuclear translocation and augmented KCNQ2/3 mRNA and IM amplitudes, suggesting the critical role of CaN and L-type channels for transcriptional regulation of M channels. Our discovery that M-channel

transcription is regulated by neuronal activity through NFAT/CaN signaling in sympathetic neurons led us to think that this may generalize throughout the nervous system to limit neuronal hyperexcitability. Thus, we measured the relative expression level of KCNQ2 and KCNQ3 transcripts in a pathological animal model of neuronal hyperexcitability, chemoconvulsant-induced seizures in mice. As the part of the brain often serving as the focal point for dangerous human seizures, we focused on the hippocampus. We used the pilocarpine as well as kainic acid (KA) convulsant-seizure models of inducing status epilepticus ( Leite et al., 2002), which corrects for any confound of altered M currents from muscarinic agonist (pilocarpine), rather than from hyperactivity.