Hypercontractility of arterial myocytes and enhanced vascular tone during diabetes are, in part, attributed to the effects of increased glucose (hyperglycemia) on L-type Ca_V1.2 channels. In murine arterial myocytes, kinase-dependent mechanisms mediate the increase in Ca_V1.2 activity in response to increased extracellular glucose. We identified a subpopulation of the Ca_V1.2 channel pore-forming subunit (α1_C) within nanometer proximity of protein kinase A (PKA) at the sarcolemma of murine and human arterial myocytes. This arrangement depended upon scaffolding of PKA by an A-kinase anchoring protein 150 (AKAP150) in mice. Glucose-mediated increases in Ca_V1.2 channel activity were associated with PKA activity, leading to α1_C phosphorylation at Ser¹⁹²⁸ Compared to arteries from low-fat diet (LFD)-fed mice and nondiabetic patients, arteries from high-fat diet (HFD)-fed mice and from diabetic patients had increased Ser¹⁹²⁸ phosphorylation and Ca_V1.2 activity. Arterial myocytes and arteries from mice lacking AKAP150 or expressing mutant AKAP150 unable to bind PKA did not exhibit increased Ser¹⁹²⁸ phosphorylation and Ca_V1.2 current density in response to increased glucose or to HFD. Consistent with a functional role for Ser¹⁹²⁸ phosphorylation, arterial myocytes and arteries from knockin mice expressing a Ca_V1.2 with Ser¹⁹²⁸ mutated to alanine (S1928A) lacked glucose-mediated increases in Ca_V1.2 activity and vasoconstriction. Furthermore, the HFD-induced increases in Ca_V1.2 current density and myogenic tone were prevented in S1928A knockin mice. These findings reveal an essential role for α1_C phosphorylation at Ser¹⁹²⁸ in stimulating Ca_V1.2 channel activity and vasoconstriction by AKAP-targeted PKA upon exposure to increased glucose and in diabetes.

The L-type Ca²⁺ channel Ca_v1.2 controls multiple functions throughout the body including heart rate and neuronal excitability. It is a key mediator of fight-or-flight stress responses triggered by a signaling pathway involving β-adrenergic receptors (βARs), cyclic adenosine monophosphate (cAMP), and protein kinase A (PKA). PKA readily phosphorylates Ser¹⁹²⁸ in Ca_v1.2 in vitro and in vivo, including in rodents and humans. However, S1928A knock-in (KI) mice have normal PKA-mediated L-type channel regulation in the heart, indicating that Ser¹⁹²⁸ is not required for regulation of cardiac Ca_v1.2 by PKA in this tissue. We report that augmentation of L-type currents by PKA in neurons was absent in S1928A KI mice. Furthermore, S1928A KI mice failed to induce long-term potentiation in response to prolonged theta-tetanus (PTT-LTP), a form of synaptic plasticity that requires Ca_v1.2 and enhancement of its activity by the β₂-adrenergic receptor (β₂AR)-cAMP-PKA cascade. Thus, there is an unexpected dichotomy in the control of Ca_v1.2 by PKA in cardiomyocytes and hippocampal neurons.

Cocaine-associated contextual cues can trigger relapse behavior by recruiting the hippocampus. Extinction of cocaine-associated contextual memories can reduce cocaine-seeking behavior, however the molecular mechanisms within the hippocampus that underlie contextual extinction behavior and subsequent reinstatement remain poorly understood. Here, we extend our previous findings for a role of Ca_v1.2 L-type Ca²⁺ channels in dopamine 1 receptor (D1R)-expressing cells in extinction of cocaine conditioned place preference (CPP) in adult male mice. We report that attenuated cocaine CPP extinction in mice lacking Ca_v1.2 channels in D1R-expressing cells (D1^cre, Ca_v1.2^fl/fl) can be rescued through chemogenetic activation of D1R-expressing cells within the dorsal dentate gyrus (dDG), but not the dorsal CA1 (dCA1). This is supported by the finding that Ca_v1.2 channels are required in excitatory cells of the dDG, but not in the dCA1, for cocaine CPP extinction. Examination of the role of S1928 phosphorylation of Ca_v1.2, a protein kinase A (PKA) site using S1928A Ca_v1.2 phosphomutant mice revealed no extinction deficit, likely due to homeostatic scaling up of extinction-dependent S845 GluA1 phosphorylation in the dDG. However, phosphomutant mice failed to show cocaine-primed reinstatement which can be reversed by chemogenetic manipulation of excitatory cells in the dDG during extinction training. These findings outline an essential role for the interaction between D1R, Ca_v1.2, and GluA1 signaling in the dDG for extinction of cocaine-associated contextual memories.

Agonist-triggered downregulation of β-adrenergic receptors (ARs) constitutes vital negative feedback to prevent cellular overexcitation. Here, we report a novel downregulation of β2AR signaling highly specific for Cav1.2. We find that β2-AR binding to Cav1.2 residues 1923-1942 is required for β-adrenergic regulation of Cav1.2. Despite the prominence of PKA-mediated phosphorylation of Cav1.2 S1928 within the newly identified β2AR binding site, its physiological function has so far escaped identification. We show that phosphorylation of S1928 displaces the β2AR from Cav1.2 upon β-adrenergic stimulation rendering Cav1.2 refractory for several minutes from further β-adrenergic stimulation. This effect is lost in S1928A knock-in mice. Although AMPARs are clustered at postsynaptic sites like Cav1.2, β2AR association with and regulation of AMPARs do not show such dissociation. Accordingly, displacement of the β2AR from Cav1.2 is a uniquely specific desensitization mechanism of Cav1.2 regulation by highly localized β2AR/cAMP/PKA/S1928 signaling. The physiological implications of this mechanism are underscored by our finding that LTP induced by prolonged theta tetanus (PTT-LTP) depends on Cav1.2 and its regulation by channel-associated β2AR.