AKAP Regulation of Neuronal L-type Calcium Channel Signaling to the Nucleus

University Of Colorado Denver, Aurora CO

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Abstract

DESCRIPTION (provided by applicant): In hippocampal neurons, somato-dendritic CaV1.2 L-type voltage-gated Ca2+ channels (LTCC) function in excitation-transcription (E-T) coupling. Depolarizing stimuli that open LTCCs activate the transcription factors cAMP-response element binding protein (CREB) and nuclear factor of activated T-cells (NFAT) through Ca2+-regulated kinase and phosphatases. Importantly, LTCC transcriptional regulation is required for long-lasting forms of excitatory synaptic plasticity that underlie learning and memory, such as late-phase long-term potentiation (L-LTP). Thus, it is crucial to understand how LTCC activity and signaling are controlled to promote efficient, specific synapse to nucleus communication. The primary question in synapse-to-nucleus signaling is how local Ca2+ signals generated in dendrites are relayed remotely to the nucleus in the soma. In the last funding period, we established the postsynaptic scaffold protein A-kinase anchoring protein (AKAP) 79/150, which binds to CaV1.2 through a modified leucine zipper (LZ) motif and anchors the cAMP-dependent protein kinase (PKA) and Ca2+-calmodulin (CaM)-activated protein phosphatase-2B (calcineurin; CaN), as an essential regulator of neuronal LTCC currents and NFAT activation. We found that AKAP-anchored PKA promoted LTCC current enhancement that was strongly opposed by a Ca2+ negative feedback loop activating AKAP-anchored CaN to favor rapid, calcium-dependent inactivation (CDI). In addition, we found that local LTCC activation of AKAP-anchored CaN was required for NFAT translocation to the nucleus and transcription in response to depolarization. However, key questions remain regarding whether AKAP79/150 regulates LTCC Ca2+ influx specifically in dendritic spines in response to glutamate receptor activation and whether postsynaptic Ca2+ signals restricted in dendrites can locally activate CaN-NFAT signaling to the nucleus. We will explore these questions using a combination of whole-cell LTCC current recordings, local glutamate uncaging, Ca2+ imaging (Aim 1), NFAT imaging (Aim 2), transcriptional analyses, and extracellular recordings of L-LTP (Aim 3). In all three aims, AKAP79/150 regulation of LTCC activity and NFAT signaling will be investigated by expressing PKA anchoring deficient (delta-PKA), CaN anchoring deficient (delta-PIX), and LZ domain (delta-LZ) AKAP79 mutants in in rat neurons or using neurons from AKAP150 delta-PIX and delta-PKA knock-in mice. Overall, this project will test a central hypothesis in synapse-to-nucleus communication that postsynaptic Ca2+ signals are locally decoded in dendrites and then efficiently relayed to the nucleus to control gene expression linked to synaptic plasticity.