{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T01:35:44Z","timestamp":1788917744078,"version":"build-2803163510"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,7,10]],"date-time":"2023-07-10T00:00:00Z","timestamp":1688947200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001652","name":"Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001652","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Netw. Physiol."],"abstract":"<jats:p>While it is well established that the isoform 2 of the hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN2) plays an important role in the development and maintenance of pain, the role of the closely related HCN4 isoform in the processing of nociceptive signals is not known. HCN4 channels are highly expressed in the thalamus, a region important for stimulus transmission and information processing. We used a brain-specific HCN4-knockout mouse line (HCN4-KO) to explore the role of HCN4 channels in acute nociceptive processing using several behavioral tests as well as a multimodal magnetic resonance imaging (MRI) approach. Functional MRI (fMRI) brain responses were measured during acute peripheral thermal stimulation complemented by resting state (RS) before and after stimulation. The data were analyzed by conventional and graph-theoretical approaches. Finally, high-resolution anatomical brain data were acquired. HCN4-KO animals showed a central thermal, but not a mechanical hypersensitivity in behavioral experiments. The open field analysis showed no significant differences in motor readouts between HCN4-KO and controls but uncovered increased anxiety in the HCN4-KO mice. Thermal stimulus-driven fMRI (s-fMRI) data revealed increased response volumes and response amplitudes for HCN4-KO, most pronounced at lower stimulation temperatures in the subcortical input, the amygdala as well as in limbic\/hippocampal regions, and in the cerebellum. These findings could be cross-validated by graph-theoretical analyses. Assessment of short-term RS before and after thermal stimulation revealed that stimulation-related modulations of the functional connectivity only occurred in control animals. This was consistent with the finding that the hippocampus was found to be smaller in HCN4-KO. In summary, the deletion of HCN4 channels impacts on processing of acute nociception, which is remarkably manifested as a thermal hypersensitive phenotype. This was mediated by the key regions hypothalamus, somatosensory cortex, cerebellum and the amygdala. As consequence, HCN4-KO mice were more anxious, and their brain-wide RS functional connectivity could not be modulated by thermal nociceptive stimulation.<\/jats:p>","DOI":"10.3389\/fnetp.2023.1090502","type":"journal-article","created":{"date-parts":[[2023,7,10]],"date-time":"2023-07-10T05:55:01Z","timestamp":1688968501000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["The impact of HCN4 channels on CNS brain networks as a new target in pain development"],"prefix":"10.3389","volume":"3","author":[{"given":"Maximilian","family":"H\u00e4fele","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silke","family":"Kreitz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Ludwig","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Hess","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel","family":"Wank","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,7,10]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"3027","DOI":"10.1111\/j.1460-9568.2004.03762.x","article-title":"Effects of ventral hippocampal lesion on thermal and mechanical nociception in neonates and adult rats","volume":"20","author":"Al Amin","year":"2004","journal-title":"Eur. J. Neurosci."},{"key":"B2","unstructured":"Allen mouse brain atlas2011"},{"key":"B3","first-page":"1","article-title":"Advanced normalization tools (ANTS)","volume":"2","author":"Avants","year":"2009","journal-title":"Insight J."},{"key":"B4","doi-asserted-by":"publisher","first-page":"1633","DOI":"10.1016\/j.neuroimage.2012.07.021","article-title":"Quantitative mouse brain phenotyping based on single and multispectral MR protocols","volume":"63","author":"Badea","year":"2012","journal-title":"Neuroimage"},{"key":"B5","doi-asserted-by":"publisher","first-page":"606","DOI":"10.1037\/h0026512","article-title":"Effects of limbic lesions on passive avoidance and reactivity to shock","volume":"66","author":"Blanchard","year":"1968","journal-title":"J. Comp. Physiol. Psychol."},{"key":"B6","doi-asserted-by":"publisher","first-page":"252","DOI":"10.1007\/s12311-008-0011-6","article-title":"Human cerebellar responses to brush and heat stimuli in healthy and neuropathic pain subjects","volume":"7","author":"Borsook","year":"2008","journal-title":"Cerebellum"},{"key":"B7","first-page":"105","volume-title":"Somatosensory cortex and tactile perceptions. Pain and touch","author":"Burton","year":"1996"},{"key":"B8","doi-asserted-by":"publisher","first-page":"7705","DOI":"10.1073\/pnas.96.14.7705","article-title":"Pain perception: Is there a role for primary somatosensory cortex?","volume":"96","author":"Bushnell","year":"1999","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B9","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1007\/BF00247291","article-title":"Mechanical response properties of ventroposterior medial thalamic neurons in the alert monkey","volume":"67","author":"Bushnell","year":"1987","journal-title":"Exp. Brain Res."},{"key":"B10","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1152\/jn.01078.2015","article-title":"Novel method for functional brain imaging in awake minimally restrained rats","volume":"116","author":"Chang","year":"2016","journal-title":"J. neurophysiology"},{"key":"B11","doi-asserted-by":"publisher","first-page":"4572","DOI":"10.3390\/ijms20184572","article-title":"Noncanonical ion channel behaviour in pain","volume":"20","author":"Ciotu","year":"2019","journal-title":"Int. J. Mol. Sci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.neulet.2016.08.021","article-title":"Inhibition of HCN channel activity in the thalamus attenuates chronic pain in rats","volume":"631","author":"Ding","year":"2016","journal-title":"Neurosci. Lett."},{"key":"B13","doi-asserted-by":"publisher","first-page":"1462","DOI":"10.1126\/science.1206243","article-title":"HCN2 ion channels play a central role in inflammatory and neuropathic pain","volume":"333","author":"Emery","year":"2011","journal-title":"Science"},{"key":"B14","doi-asserted-by":"publisher","first-page":"3270","DOI":"10.1152\/jn.90777.2008","article-title":"The global signal and observed anticorrelated resting state brain networks","volume":"101","author":"Fox","year":"2009","journal-title":"J. neurophysiology"},{"key":"B15","volume-title":"The mouse brain in stereotaxic coordinates","author":"Franklin","year":"2008"},{"key":"B16","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1007\/s00441-009-0904-9","article-title":"Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice","volume":"339","author":"Fried","year":"2010","journal-title":"Cell Tissue Res."},{"key":"B17","doi-asserted-by":"publisher","first-page":"e12550","DOI":"10.1111\/gbb.12550","article-title":"HCN4 knockdown in dorsal hippocampus promotes anxiety-like behavior in mice","volume":"18","author":"Gunther","year":"2019","journal-title":"Genes Brain Behav."},{"key":"B18","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1016\/j.neuroimage.2015.10.009","article-title":"Morphological maturation of the mouse brain: An in vivo MRI and histology investigation","volume":"125","author":"Hammelrath","year":"2016","journal-title":"Neuroimage"},{"key":"B19","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1016\/0304-3959(88)90026-7","article-title":"A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia","volume":"32","author":"Hargreaves","year":"1988","journal-title":"Pain"},{"key":"B20","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.pneurobio.2013.10.001","article-title":"Neurophysiology of HCN channels: From cellular functions to multiple regulations","volume":"112","author":"He","year":"2014","journal-title":"Prog. Neurobiol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1038\/nn0504-483","article-title":"Gene expression nervous system atlas (GENSAT)","volume":"7","author":"Heintz","year":"2004","journal-title":"Nat. Neurosci."},{"key":"B22","doi-asserted-by":"publisher","first-page":"386","DOI":"10.1007\/s00381-002-0604-1","article-title":"Functional anatomy of thalamus and basal ganglia","volume":"18","author":"Herrero","year":"2002","journal-title":"Childs Nerv. Syst."},{"key":"B23","doi-asserted-by":"publisher","first-page":"2012","DOI":"10.1097\/j.pain.0000000000001005","article-title":"Protein kinase A regulates inflammatory pain sensitization by modulating HCN2 channel activity in nociceptive sensory neurons","volume":"158","author":"Herrmann","year":"2017","journal-title":"Pain"},{"key":"B24","doi-asserted-by":"publisher","first-page":"3731","DOI":"10.1073\/pnas.1011774108","article-title":"Blockade of TNF-\u03b1 rapidly inhibits pain responses in the central nervous system","volume":"108","author":"Hess","year":"2011","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B25","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1016\/j.ejpain.2006.01.005","article-title":"Imaging of hyperalgesia in rats by functional MRI","volume":"11","author":"Hess","year":"2007","journal-title":"Eur. J. Pain"},{"key":"B26","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1016\/j.neuroscience.2013.01.028","article-title":"HCN4 subunit expression in fast-spiking interneurons of the rat spinal cord and hippocampus","volume":"237","author":"Hughes","year":"2013","journal-title":"Neuroscience"},{"key":"B27","first-page":"132","article-title":"The effects of morphine methadone and meperidine on some reflex responses of spinal animals to nociceptive stimulation","volume":"101","author":"Irwin","year":"1951","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"B28","doi-asserted-by":"publisher","first-page":"334","DOI":"10.3389\/fnins.2018.00334","article-title":"A new analysis of resting state connectivity and graph theory reveals distinctive short-term modulations due to whisker stimulation in rats","volume":"12","author":"Kreitz","year":"2018","journal-title":"Front. Neurosci."},{"key":"B29","doi-asserted-by":"publisher","first-page":"1480","DOI":"10.1097\/00000542-199812000-00026","article-title":"Local cerebral blood flow, local cerebral glucose utilization, and flow-metabolism coupling during sevoflurane versus isoflurane anesthesia in rats","volume":"89","author":"Lenz","year":"1998","journal-title":"Anesthesiology"},{"key":"B30","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/s1058-9139(06)80011-2","article-title":"The ventral posterior nucleus of thalamus is involved in the generation of central pain syndromes","volume":"1","author":"Lenz","year":"1992","journal-title":"APS J."},{"key":"B31","doi-asserted-by":"publisher","first-page":"2323","DOI":"10.1093\/emboj\/18.9.2323","article-title":"Two pacemaker channels from human heart with profoundly different activation kinetics","volume":"18","author":"Ludwig","year":"1999","journal-title":"EMBO J."},{"key":"B32","doi-asserted-by":"publisher","first-page":"1477","DOI":"10.1016\/j.neuroscience.2006.10.048","article-title":"Role of peripheral hyperpolarization-activated cyclic nucleotide-modulated channel pacemaker channels in acute and chronic pain models in the rat","volume":"144","author":"Luo","year":"2007","journal-title":"Neuroscience"},{"key":"B33","doi-asserted-by":"publisher","first-page":"242","DOI":"10.1111\/j.1460-9568.2009.06812.x","article-title":"Dose-dependent effect of isoflurane on neurovascular coupling in rat cerebral cortex","volume":"30","author":"Masamoto","year":"2009","journal-title":"Eur. J. Neurosci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"265","DOI":"10.1111\/j.1749-6632.2001.tb05830.x","article-title":"Plasticity of the hippocampus: Adaptation to chronic stress and allostatic load","volume":"933","author":"McEwen","year":"2001","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"B35","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1016\/s0169-328x(00)00155-8","article-title":"Cloning and localization of the hyperpolarization-activated cyclic nucleotide-gated channel family in rat brain","volume":"81","author":"Monteggia","year":"2000","journal-title":"Brain Res. Mol. Brain Res."},{"key":"B36","doi-asserted-by":"publisher","first-page":"975","DOI":"10.1515\/BC.1999.121","article-title":"Differential distribution of four hyperpolarization-activated cation channels in mouse brain","volume":"380","author":"Moosmang","year":"1999","journal-title":"Biol. Chem."},{"key":"B37","doi-asserted-by":"publisher","first-page":"3795","DOI":"10.1523\/JNEUROSCI.6709-10.2011","article-title":"Aversion-related circuitry in the cerebellum: Responses to noxious heat and unpleasant images","volume":"31","author":"Moulton","year":"2011","journal-title":"J. Neurosci. official J. Soc. Neurosci."},{"key":"B38","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1016\/j.brainresrev.2010.05.005","article-title":"The cerebellum and pain: Passive integrator or active participator?","volume":"65","author":"Moulton","year":"2010","journal-title":"Brain Res. Rev."},{"key":"B39","doi-asserted-by":"publisher","first-page":"1749","DOI":"10.1113\/jphysiol.2012.247361","article-title":"Hyperpolarisation-activated cyclic nucleotide-gated channels regulate the spontaneous firing rate of olfactory receptor neurons and affect glomerular formation in mice","volume":"591","author":"Nakashima","year":"2013","journal-title":"J. Physiol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"628","DOI":"10.1016\/j.cell.2010.09.047","article-title":"A genome-wide Drosophila screen for heat nociception identifies \u03b12\u03b43 as an evolutionarily conserved pain gene","volume":"143","author":"Neely","year":"2010","journal-title":"Cell"},{"key":"B41","doi-asserted-by":"publisher","first-page":"952","DOI":"10.1016\/j.bbrc.2010.12.087","article-title":"HCN channel activity-dependent modulation of inhibitory synaptic transmission in the rat basolateral amygdala","volume":"404","author":"Park","year":"2011","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"B42","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1146\/annurev.physiol.65.092101.142734","article-title":"Hyperpolarization-activated cation currents: From molecules to physiological function","volume":"65","author":"Robinson","year":"2003","journal-title":"Annu. Rev. Physiol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"5264","DOI":"10.1523\/JNEUROSCI.20-14-05264.2000","article-title":"Molecular and functional heterogeneity of hyperpolarization-activated pacemaker channels in the mouse CNS","volume":"20","author":"Santoro","year":"2000","journal-title":"J. Neurosci."},{"key":"B44","doi-asserted-by":"publisher","first-page":"1079","DOI":"10.1016\/j.pain.2014.02.006","article-title":"HCN2 channels account for mechanical (but not heat) hyperalgesia during long-standing inflammation","volume":"155","author":"Schnorr","year":"2014","journal-title":"PAIN\u00ae."},{"key":"B45","doi-asserted-by":"publisher","first-page":"9391","DOI":"10.1073\/pnas.96.16.9391","article-title":"Molecular characterization of a slowly gating human hyperpolarization-activated channel predominantly expressed in thalamus, heart, and testis","volume":"96","author":"Seifert","year":"1999","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B46","doi-asserted-by":"publisher","first-page":"e0120345","DOI":"10.1371\/journal.pone.0120345","article-title":"Default mode network, motor network, dorsal and ventral basal ganglia networks in the rat brain: Comparison to human networks using resting state-fMRI","volume":"10","author":"Sierakowiak","year":"2015","journal-title":"PLoS One"},{"key":"B47","doi-asserted-by":"publisher","first-page":"15235","DOI":"10.1073\/pnas.2434235100","article-title":"The hyperpolarization-activated channel HCN4 is required for the generation of pacemaker action potentials in the embryonic heart","volume":"100","author":"Stieber","year":"2003","journal-title":"Proc. Natl. Acad. Sci."},{"key":"B48","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1016\/s0014-2999(00)00569-0","article-title":"Noradrenaline systems in the hypothalamus, amygdala and locus coeruleus are involved in the provocation of anxiety: Basic studies","volume":"405","author":"Tanaka","year":"2000","journal-title":"Eur. J. Pharmacol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1016\/s0304-3959(98)00184-5","article-title":"The cortical representation of pain","volume":"79","author":"Treede","year":"1999","journal-title":"Pain"},{"key":"B50","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1038\/12703","article-title":"Disruption of the glucocorticoid receptor gene in the nervous system results in reduced anxiety","volume":"23","author":"Tronche","year":"1999","journal-title":"Nat. Genet."},{"key":"B51","doi-asserted-by":"publisher","first-page":"743","DOI":"10.1002\/hbm.20902","article-title":"Distinct and shared cerebral activations in processing innocuous versus noxious contact heat revealed by functional magnetic resonance imaging","volume":"31","author":"Tseng","year":"2010","journal-title":"Hum. Brain Mapp."},{"key":"B52","doi-asserted-by":"publisher","first-page":"470","DOI":"10.1007\/s00018-008-8525-0","article-title":"HCN channels: Structure, cellular regulation and physiological function","volume":"66","author":"Wahl-Schott","year":"2009","journal-title":"Cell Mol. Life Sci."},{"key":"B53","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/j.pain.2010.02.023","article-title":"Abnormal thalamocortical activity in patients with complex regional pain syndrome (CRPS) type I","volume":"150","author":"Walton","year":"2010","journal-title":"Pain"},{"key":"B54","doi-asserted-by":"publisher","first-page":"e0266669","DOI":"10.1371\/journal.pone.0266669","article-title":"Imaging the influence of peripheral TRPV1-signaling on cerebral nociceptive processing applying fMRI-based graph theory in a resiniferatoxin rat model","volume":"17","author":"Wank","year":"2022","journal-title":"PLoS One"},{"key":"B55","doi-asserted-by":"publisher","first-page":"204","DOI":"10.1016\/j.brainres.2019.03.036","article-title":"HCN channel antagonist ZD7288 ameliorates neuropathic pain and associated depression","volume":"1717","author":"Yang","year":"2019","journal-title":"Brain Res."},{"key":"B56","doi-asserted-by":"publisher","first-page":"1197","DOI":"10.1016\/j.neuroimage.2010.06.041","article-title":"Network-based statistic: Identifying differences in brain networks","volume":"53","author":"Zalesky","year":"2010","journal-title":"NeuroImage"},{"key":"B57","doi-asserted-by":"publisher","first-page":"2291","DOI":"10.1093\/cercor\/bhz047","article-title":"The hyperpolarization-activated HCN4 channel is important for proper maintenance of oscillatory activity in the thalamocortical system","volume":"29","author":"Zobeiri","year":"2019","journal-title":"Cereb. cortex"},{"key":"B58","doi-asserted-by":"publisher","first-page":"1429","DOI":"10.1016\/j.bbrc.2016.08.141","article-title":"Expression and cellular localization of HCN channels in rat cerebellar granule neurons","volume":"478","author":"Zuniga","year":"2016","journal-title":"Biochem. Biophys. Res. Commun."}],"container-title":["Frontiers in Network Physiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnetp.2023.1090502\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,10]],"date-time":"2023-07-10T05:55:35Z","timestamp":1688968535000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnetp.2023.1090502\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,10]]},"references-count":58,"alternative-id":["10.3389\/fnetp.2023.1090502"],"URL":"https:\/\/doi.org\/10.3389\/fnetp.2023.1090502","relation":{},"ISSN":["2674-0109"],"issn-type":[{"value":"2674-0109","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,10]]},"article-number":"1090502"}}