Interaction Between HCN and Slack Channels Regulates mPFC Pyramidal Cell Excitability and Working Memory

The ability of monkeys and rats to carry out spatial working memory tasks has been shown to depend on the persistent firing of pyramidal cells in the prefrontal cortex (PFC), arising from recurrent excitatory connections on dendritic spines. These spines express hyperpolarization-activated cyclic nucleotide-gated (HCN) channels whose open state is increased by cAMP signaling, and which markedly alter PFC network connectivity and neuronal firing. In traditional neural circuits, activation of these non-selective cation channels leads to neuronal depolarization and increased firing rate. Paradoxically, cAMP activation of HCN channels in PFC pyramidal cells reduces working memory-related neuronal firing. This suggests that activation of HCN channels may hyperpolarize rather than depolarize these neurons. The current study tested the hypothesis that Na+ influx through HCN channels activates Na+-activated K+ (KNa or Slack) channels to hyperpolarize the membrane. We have found that HCN and Slack KNa channels co-immunoprecipitate in cortical extracts and that, by immunoelectron microscopy, they colocalize at postsynaptic spines of PFC pyramidal neurons. A specific blocker of HCN channels, ZD7288, reduces KNa current in pyramidal cells that express both HCN and Slack channels, indicating that blockade of HCN channels reduced K+ current indirectly by lowering Na+ influx. In contrast, ZD7288 has no effect on KNa currents in an HEK cell line stably expressing this Slack channels but no HCN channels, demonstrating that ZD7288 does not block Slack channels directly. Activation of HCN channels by cAMP in a cell line expressing a Ca2+ reporter results in elevation of cytoplasmic Ca2+, but the effect of cAMP is completely reversed if the HCN channels are co-expressed with Slack channels. Finally, we have used a novel pharmacological blocker of Slack channels to show that inhibition of either Slack or HCN channels in rat PFC improves working memory performance, and that the actions of Slack and HCN channel blockers occlude each other in the memory task. Our results suggest that the regulation of working memory by HCN channels in PFC pyramidal neurons is mediated by an HCN-Slack channel complex that links activation HCN channels to suppression of neuronal excitability.

[1]  Yu Du,et al.  Structure-activity relationship studies in a new series of 2-amino-N-phenylacetamide inhibitors of Slack potassium channels. , 2022, Bioorganic & medicinal chemistry letters.

[2]  Tracy S. Gertler,et al.  KNa1.1 gain-of-function preferentially dampens excitability of murine parvalbumin-positive interneurons , 2022, Neurobiology of Disease.

[3]  Z. Hughes,et al.  Discovery of the First Orally Available, Selective KNa1.1 Inhibitor: In Vitro and In Vivo Activity of an Oxadiazole Series. , 2021, ACS medicinal chemistry letters.

[4]  T. Hughes,et al.  Optically activated, customizable, excitable cells , 2020, PloS one.

[5]  A. Arnsten,et al.  The genie in the bottle-magnified calcium signaling in dorsolateral prefrontal cortex , 2020, Molecular Psychiatry.

[6]  William F Tobin,et al.  Reduced GABAergic Neuron Excitability, Altered Synaptic Connectivity, and Seizures in a KCNT1 Gain-of-Function Mouse Model of Childhood Epilepsy , 2020, Cell reports.

[7]  C. Weaver,et al.  VU0606170, a Selective Slack Channels Inhibitor, Decreases Calcium Oscillations in Cultured Cortical Neurons. , 2020, ACS chemical neuroscience.

[8]  C. Fishwick,et al.  Structure-Based Identification and Characterization of Inhibitors of the Epilepsy-Associated KNa1.1 (KCNT1) Potassium Channel , 2020, iScience.

[9]  J. Mazer,et al.  Core Differences in Synaptic Signaling Between Primary Visual and Dorsolateral Prefrontal Cortex , 2018, Cerebral cortex.

[10]  Erik C. Nook,et al.  Weak dorsolateral prefrontal response to social criticism predicts worsened mood and symptoms following social conflict in people at familial risk for schizophrenia , 2018, NeuroImage: Clinical.

[11]  D. Brager,et al.  Perisomatic changes in h-channels regulate depressive behaviors following chronic unpredictable stress , 2017, Molecular Psychiatry.

[12]  P. Licznerski,et al.  Inhibition of Bcl-xL prevents pro-death actions of ΔN-Bcl-xL at the mitochondrial inner membrane during glutamate excitotoxicity , 2017, Cell Death and Differentiation.

[13]  C. Elger,et al.  Mesial temporal lobe epilepsy associated with KCNT1 mutation , 2017, Seizure.

[14]  Leonard K. Kaczmarek,et al.  International Union of Basic and Clinical Pharmacology. C. Nomenclature and Properties of Calcium-Activated and Sodium-Activated Potassium Channels , 2017, Pharmacological Reviews.

[15]  C. Berridge,et al.  Stress Degrades Prefrontal Cortex Neuronal Coding of Goal‐Directed Behavior , 2016, Cerebral cortex.

[16]  D. P. Jenkins,et al.  Stimulation of Slack K(+) Channels Alters Mass at the Plasma Membrane by Triggering Dissociation of a Phosphatase-Regulatory Complex. , 2016, Cell reports.

[17]  A. Weisz,et al.  Characterization of two de novo KCNT1 mutations in children with malignant migrating partial seizures in infancy , 2016, Molecular and Cellular Neuroscience.

[18]  A. Arnsten,et al.  Stress weakens prefrontal networks: molecular insults to higher cognition , 2015, Nature Neuroscience.

[19]  T. Shiihara,et al.  De novo KCNT1 mutations in early‐onset epileptic encephalopathy , 2015, Epilepsia.

[20]  G. Carvill,et al.  Mutations in KCNT1 cause a spectrum of focal epilepsies , 2015, Epilepsia.

[21]  Daeyeol Lee,et al.  Stress Impairs Prefrontal Cortical Function Via D1 Dopamine Receptor Interactions With HCN Channels , 2015 .

[22]  L. Kaczmarek,et al.  Human slack potassium channel mutations increase positive cooperativity between individual channels. , 2014, Cell reports.

[23]  H. Oguni,et al.  A novel KCNT1 mutation in a Japanese patient with epilepsy of infancy with migrating focal seizures , 2014, Human Genome Variation.

[24]  Boris Yamrom,et al.  The contribution of de novo coding mutations to autism spectrum disorder , 2014, Nature.

[25]  E. Chuang,et al.  Disease-Targeted Sequencing of Ion Channel Genes identifies de novo mutations in Patients with Non-Familial Brugada Syndrome , 2014, Scientific Reports.

[26]  L. Kaczmarek,et al.  Emerging role of the KCNT1 Slack channel in intellectual disability , 2014, Front. Cell. Neurosci..

[27]  A. Suominen,et al.  Epilepsy Among Children and Adolescents with Autism Spectrum Disorders: A Population-Based Study , 2014, Journal of autism and developmental disorders.

[28]  I. Scheffer,et al.  KCNT1 gain of function in 2 epilepsy phenotypes is reversed by quinidine , 2014, Annals of neurology.

[29]  Zhian Hu,et al.  Neurophysiology of HCN channels: From cellular functions to multiple regulations , 2014, Progress in Neurobiology.

[30]  Peter Donnelly,et al.  Clinical whole-genome sequencing in severe early-onset epilepsy reveals new genes and improves molecular diagnosis , 2014, Human molecular genetics.

[31]  A. Vanderver,et al.  Identification of a novel de novo p.Phe932Ile KCNT1 mutation in a patient with leukoencephalopathy and severe epilepsy. , 2014, Pediatric neurology.

[32]  M. Ōsawa,et al.  A recurrent KCNT1 mutation in two sporadic cases with malignant migrating partial seizures in infancy. , 2013, Gene.

[33]  Christine M Constantinople,et al.  Prefrontal Cortex HCN1 Channels Enable Intrinsic Persistent Neural Firing and Executive Memory Function , 2013, The Journal of Neuroscience.

[34]  A. Arnsten,et al.  Constellation of HCN channels and cAMP regulating proteins in dendritic spines of the primate prefrontal cortex: potential substrate for working memory deficits in schizophrenia. , 2013, Cerebral cortex.

[35]  I. Scheffer,et al.  Migrating partial seizures of infancy: expansion of the electroclinical, radiological and pathological disease spectrum. , 2013, Brain : a journal of neurology.

[36]  L. Kaczmarek,et al.  De novo gain-of-function KCNT1 channel mutations cause malignant migrating partial seizures of infancy , 2012, Nature Genetics.

[37]  Katherine R. Smith,et al.  Missense mutations in the sodium-gated potassium channel gene KCNT1 cause severe autosomal dominant nocturnal frontal lobe epilepsy , 2012, Nature Genetics.

[38]  Daeyeol Lee,et al.  Neuronal basis of age-related working memory decline , 2011, Nature.

[39]  Akihiro Yasuhara,et al.  Correlation between EEG abnormalities and symptoms of autism spectrum disorder (ASD) , 2010, Brain and Development.

[40]  L. Kaczmarek,et al.  The slack sodium-activated potassium channel provides a major outward current in olfactory neurons of Kv1.3-/- super-smeller mice. , 2010, Journal of neurophysiology.

[41]  A. Bhattacharjee,et al.  cAMP-dependent kinase does not modulate the Slack sodium-activated potassium channel , 2009, Neuropharmacology.

[42]  L. Kaczmarek,et al.  Amino‐termini isoforms of the Slack K+ channel, regulated by alternative promoters, differentially modulate rhythmic firing and adaptation , 2008, The Journal of physiology.

[43]  M. Baulac,et al.  Epilepsy in Autism is Associated with Intellectual Disability and Gender: Evidence from a Meta-Analysis , 2008, Biological Psychiatry.

[44]  Murrey G. Olmsted,et al.  Co‐occurring conditions associated with FMR1 gene variations: Findings from a national parent survey , 2008, American journal of medical genetics. Part A.

[45]  D. McCormick,et al.  α2A-Adrenoceptors Strengthen Working Memory Networks by Inhibiting cAMP-HCN Channel Signaling in Prefrontal Cortex , 2007, Cell.

[46]  L. Kaczmarek,et al.  Pharmacological activation and inhibition of Slack (Slo2.2) channels , 2006, Neuropharmacology.

[47]  D. Surmeier,et al.  Dendritic Excitability of Mouse Frontal Cortex Pyramidal Neurons Is Shaped by the Interaction among HCN, Kir2, and Kleak Channels , 2005, The Journal of Neuroscience.

[48]  Matthew F. Nolan,et al.  A Behavioral Role for Dendritic Integration HCN1 Channels Constrain Spatial Memory and Plasticity at Inputs to Distal Dendrites of CA1 Pyramidal Neurons , 2004, Cell.

[49]  R. Shigemoto,et al.  Immunohistochemical localization of Ih channel subunits, HCN1–4, in the rat brain , 2004, The Journal of comparative neurology.

[50]  L. Kaczmarek,et al.  Slick (Slo2.1), a Rapidly-Gating Sodium-Activated Potassium Channel Inhibited by ATP , 2003, The Journal of Neuroscience.

[51]  A. Arnsten,et al.  Dysregulation of Protein Kinase A Signaling in the Aged Prefrontal Cortex New Strategy for Treating Age-Related Cognitive Decline , 2003, Neuron.

[52]  L. Kaczmarek,et al.  The Sodium-Activated Potassium Channel Is Encoded by a Member of the Slo Gene Family , 2003, Neuron.

[53]  L. Kaczmarek,et al.  Localization of the Slack potassium channel in the rat central nervous system , 2002, The Journal of comparative neurology.

[54]  Gábor Tamás,et al.  Polarized and compartment-dependent distribution of HCN1 in pyramidal cell dendrites , 2002, Nature Neuroscience.

[55]  M. Lauritsen,et al.  The genetics of autism , 2001, Acta psychiatrica Scandinavica.

[56]  S. Siegelbaum,et al.  Properties of Hyperpolarization-Activated Pacemaker Current Defined by Coassembly of Hcn1 and Hcn2 Subunits and Basal Modulation by Cyclic Nucleotide , 2001, The Journal of general physiology.

[57]  P. Bosco,et al.  Epilepsy and EEG Findings in Males with Fragile X Syndrome , 1999, Epilepsia.

[58]  L. Kaczmarek,et al.  Formation of intermediate-conductance calcium-activated potassium channels by interaction of Slack and Slo subunits , 1998, Nature Neuroscience.

[59]  P. Goldman-Rakic.  Cellular basis of working memory , 1995, Neuron.

[60]  P. Goldman-Rakic,et al.  Mnemonic coding of visual space in the monkey's dorsolateral prefrontal cortex. , 1989, Journal of neurophysiology.