{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T16:07:45Z","timestamp":1753891665280,"version":"3.41.2"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,2,16]],"date-time":"2024-02-16T00:00:00Z","timestamp":1708041600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:p>Protein kinase C \u03b3 (PKC\u03b3), a neuronal isoform present exclusively in the central nervous system, is most abundantly expressed in cerebellar Purkinje cells (PCs). Targeted deletion of PKC\u03b3 causes a climbing fiber synapse elimination in developing PCs and motor deficit. However, physiological roles of PKC\u03b3 in adult mouse PCs are little understood. In this study, we aimed to unravel the roles of PKC\u03b3 in mature mouse PCs by deleting PKC\u03b3 from adult mouse PCs of PKC\u03b3<jats:italic><jats:sup>fl\/fl<\/jats:sup><\/jats:italic> mice via cerebellar injection of adeno-associated virus (AAV) vectors expressing Cre recombinase under the control of the PC-specific L7-6 promoter. Whole cell patch-clamp recording of PCs showed higher intrinsic excitability in PCs virally lacking PKC\u03b3 [PKC\u03b3-conditional knockout (PKC\u03b3-cKO) PCs] than in wild-type (WT) mouse PCs in the zebrin-negative module, but not in the zebrin-positive module. AAV-mediated PKC\u03b3 re-expression in PKC\u03b3-deficient mouse PCs in the zebrin-negative module restored the enhanced intrinsic excitability to a level comparable to that of wild-type mouse PCs. In parallel with higher intrinsic excitability, we found larger hyperpolarization-activated cyclic nucleotide-gated (HCN) channel currents in PKC\u03b3-cKO PCs located in the zebrin-negative module, compared with those in WT mouse PCs in the same region. However, pharmacological inhibition of the HCN currents did not restore the enhanced intrinsic excitability in PKC\u03b3-cKO PCs in the zebrin-negative module. These results suggested that PKC\u03b3 suppresses the intrinsic excitability in zebrin-negative PCs, which is likely independent of the HCN current inhibition.<\/jats:p>","DOI":"10.3389\/fncel.2024.1349878","type":"journal-article","created":{"date-parts":[[2024,2,16]],"date-time":"2024-02-16T04:34:31Z","timestamp":1708058071000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Protein kinase C\u03b3 negatively regulates the intrinsic excitability in zebrin-negative cerebellar Purkinje cells"],"prefix":"10.3389","volume":"18","author":[{"given":"Masashi","family":"Watanave","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mika","family":"Kawachi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ayumu","family":"Konno","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryo","family":"Aoki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuuki","family":"Fukai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yasunori","family":"Matsuzaki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryosuke","family":"Kaneko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hirokazu","family":"Hirai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2024,2,16]]},"reference":[{"key":"B1","doi-asserted-by":"crossref","first-page":"1253","DOI":"10.1016\/0092-8674(93)90613-U","article-title":"Modified hippocampal long-term potentiation in PKCy-mutant mice.","volume":"75","author":"Abeliovich","year":"","journal-title":"Cell"},{"key":"B2","doi-asserted-by":"publisher","first-page":"1263","DOI":"10.1016\/0092-8674(93)90614-v","article-title":"PKC\u03b3 mutant mice exhibit mild deficits in spatial and contextual learning.","volume":"75","author":"Abeliovich","year":"","journal-title":"Cell"},{"key":"B3","doi-asserted-by":"crossref","first-page":"19854","DOI":"10.1074\/jbc.M801492200","article-title":"Enzymological analysis of mutant protein kinase C\u03b3 causing spinocerebellar ataxia type 14 and dysfunction in Ca2+ homeostasis.","volume":"283","author":"Adachi","year":"2008","journal-title":"J. Biol. Chem."},{"key":"B4","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/0092-8674(94)90205-4","article-title":"Deficient cerebellar long-term depression and impaired motor learning in mGluR1 mutant mice.","volume":"79","author":"Aiba","year":"1994","journal-title":"Cell"},{"key":"B5","doi-asserted-by":"crossref","DOI":"10.1523\/JNEUROSCI.08-10-03850.1988","article-title":"Distinct cellular expression of \u03b2I-and \u03b2ll-subspecies of protein kinase C in rat cerebellum.","volume":"8","author":"Ase","year":"1988","journal-title":"J. Neurosci."},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3390\/cells12040623","article-title":"Lobule-related action potential shape- and history-dependent current integration in Purkinje cells of adult and developing mice.","volume":"12","author":"Beekhof","year":"2023","journal-title":"Cells"},{"key":"B7","doi-asserted-by":"publisher","first-page":"13630","DOI":"10.1523\/JNEUROSCI.3226-10.2010","article-title":"Intrinsic plasticity complements long-term potentiation in parallel fiber input gain control in cerebellar Purkinje cells.","volume":"30","author":"Belmeguenai","year":"2010","journal-title":"J. Neurosci."},{"key":"B8","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1002\/cne.902910405","article-title":"Zebrin II: a polypeptide antigen expressed selectively by purkinje cells reveals compartments in rat and fish cerebellum.","volume":"291","author":"Brochu","year":"1990","journal-title":"J. Compar. Neurol."},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.42766","article-title":"HCN channel-mediated neuromodulation can control action potential velocity and fidelity in central axons.","volume":"8","author":"Byczkowicz","year":"2019","journal-title":"Elife"},{"key":"B10","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1038\/nrn3886","article-title":"Redefining the cerebellar cortex as an assembly of non-uniform Purkinje cell microcircuits.","volume":"16","author":"Cerminara","year":"2015","journal-title":"Nat. Rev. Neurosci."},{"key":"B11","doi-asserted-by":"publisher","first-page":"1172","DOI":"10.1038\/nn.4593","article-title":"Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems.","volume":"20","author":"Chan","year":"2017","journal-title":"Nat. Neurosci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1233","DOI":"10.1016\/0092-8674(95)90148-5","article-title":"Impaired motor coordination correlates with persistent multiple climbing fiber innervation in PKC\u03b3 mutant mice.","volume":"83","author":"Chen","year":"1995","journal-title":"Cell"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3000596","article-title":"SK2 channels in cerebellar Purkinje cells contribute to excitability modulation in motor-learning\u2013specific memory traces.","volume":"18","author":"Grasselli","year":"2020","journal-title":"PLoS Biol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"458","DOI":"10.1038\/mp.2016.99","article-title":"HCN-channel dendritic targeting requires bipartite interaction with TRIP8b and regulates antidepressant-like behavioral effects.","volume":"22","author":"Han","year":"2017","journal-title":"Mol. Psychiatry"},{"key":"B15","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1016\/j.coph.2018.02.002","article-title":"Type 1 metabotropic glutamate receptor and its signaling molecules as therapeutic targets for the treatment of cerebellar disorders.","volume":"38","author":"Hirai","year":"2018","journal-title":"Curr. Opin. Pharmacol."},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1038\/srep40257","article-title":"HCN1 channels reduce the rate of exocytosis from a subset of cortical synaptic terminals.","volume":"7","author":"Huang","year":"2017","journal-title":"Sci. Rep."},{"key":"B17","doi-asserted-by":"crossref","first-page":"1832","DOI":"10.1126\/science.288.5472.1832","article-title":"mGluR1 in cerebellar Purkinje cells essential for long-term depression, synapse elimination, and motor coordination.","volume":"208","author":"Ichise","year":"2000","journal-title":"Science"},{"key":"B18","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1002\/cne.24343","article-title":"Inhibitory neuron-specific Cre-dependent red fluorescent labeling using VGAT BAC-based transgenic mouse lines with identified transgene integration sites.","volume":"526","author":"Kaneko","year":"2018","journal-title":"J. Comp. Neurol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1223","DOI":"10.1016\/0092-8674(95)90147-7","article-title":"Impaired synapse elimination during cerebellar development in PKC\u03b3 mutant mice.","volume":"83","author":"Kano","year":"1995","journal-title":"Cell"},{"key":"B20","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1016\/s0896-6273(01)80047-7","article-title":"Persistent multiple climbing fiber innervationof cerebellar purkinje cellsin mice lacking mGluR1.","volume":"18","author":"Kano","year":"1997","journal-title":"Neuron"},{"key":"B21","doi-asserted-by":"publisher","first-page":"2173","DOI":"10.1098\/rstb.2008.2270","article-title":"Type-1 metabotropic glutamate receptor in cerebellar Purkinje cells: a key molecule responsible for long-term depression, endocannabinoid signalling and synapse elimination.","volume":"363","author":"Kano","year":"2008","journal-title":"Philos. Trans. R. Soc. B Biol. Sci."},{"key":"B22","doi-asserted-by":"crossref","first-page":"15724","DOI":"10.1073\/pnas.95.26.15724","article-title":"Phospholipase C\u03b24 is specifically involved in climbing fiber synapse elimination in the developping cerebellum.","volume":"95","author":"Kano","year":"1998","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B23","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1038\/38508","article-title":"Phospholipase C isozymes selectively couple to specific neurotransmitter receptors.","volume":"389","author":"Kim","year":"1997","journal-title":"Nature"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2020.108914","article-title":"Efficient whole brain transduction by systemic infusion of minimally purified AAV-PHP.eB.","volume":"346","author":"Konno","year":"2020","journal-title":"J. Neurosci. Methods"},{"key":"B25","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1126\/science.278.5336.279","article-title":"Preserved acute pain and reduced neuropathic pain in mice lacking PKCgamma.","volume":"278","author":"Malmberg","year":"1997","journal-title":"Science"},{"key":"B26","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1016\/j.jneumeth.2013.12.004","article-title":"Generation of a neurodegenerative disease mouse model using lentiviral vectors carrying an enhanced synapsin I promoter.","volume":"223","author":"Matsuzaki","year":"2014","journal-title":"J. Neurosci. Methods"},{"key":"B27","doi-asserted-by":"crossref","first-page":"1945","DOI":"10.1046\/j.0953-816x.2001.01570.x","article-title":"Deficient long-term synaptic depression in the rostral cerebellum correlated with impaired motor learning in phospholipase C \u03b24 mutant mice.","volume":"13","author":"Miyata","year":"2001","journal-title":"Eur. J. Neurosci."},{"key":"B28","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1016\/j.omtm.2017.07.006","article-title":"Minimal Purkinje cell-specific PCP2\/L7 promoter virally available for rodents and non-human primates.","volume":"6","author":"Nitta","year":"2017","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"B29","doi-asserted-by":"publisher","first-page":"532","DOI":"10.1159\/000350074","article-title":"Protein kinase C activation inhibits rat and human hyperpolarization activated cyclic nucleotide gated channel (HCN)1-mediated current in mammalian cells.","volume":"31","author":"Reetz","year":"2013","journal-title":"Cell. Physiol. Biochem."},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-15791-y","article-title":"An axon-specific expression of HCN channels catalyzes fast action potential signaling in GABAergic interneurons.","volume":"11","author":"Roth","year":"2020","journal-title":"Nat. Commun."},{"key":"B31","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1093\/oxfordjournals.jbchem.a003274","article-title":"Protein Kinase C (PKC): function of neuron specific isotype.","volume":"132","author":"Saito","year":"2002","journal-title":"J. Biochem."},{"key":"B32","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1002\/cne.20912","article-title":"Complementary stripes of phospholipase C\u03b23 and C\u03b24 expression by Purkinje cell subsets in the mouse cerebellum.","volume":"496","author":"Sarna","year":"2006","journal-title":"J. Comp. Neurol."},{"key":"B33","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1007\/s12311-010-0161-1","article-title":"High transgene expression by lentiviral vectors causes maldevelopment of Purkinje cells in vivo.","volume":"9","author":"Sawada","year":"2010","journal-title":"Cerebellum"},{"key":"B34","doi-asserted-by":"publisher","first-page":"660","DOI":"10.1111\/jnc.14239","article-title":"Regulatory connection between the expression level of classical protein kinase C and pruning of climbing fibers from cerebellar Purkinje cells.","volume":"143","author":"Takahashi","year":"2017","journal-title":"J. Neurochem."},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2021.103705","article-title":"Heterogeneity of intrinsic plasticity in cerebellar Purkinje cells linked with cortical molecular zones.","volume":"25","author":"Viet","year":"2022","journal-title":"iScience"},{"key":"B36","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1038\/nn1539","article-title":"Patterned expression of Purkinje cell glutamate transporters controls synaptic plasticity.","volume":"8","author":"Wadiche","year":"2005","journal-title":"Nat. Neurosci."},{"key":"B37","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1016\/j.nbd.2018.10.014","article-title":"Pharmacological enhancement of retinoid-related orphan receptor \u03b1 function mitigates spinocerebellar ataxia type 3 pathology.","volume":"121","author":"Watanave","year":"2019","journal-title":"Neurobiol. Dis."},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2018.00490","article-title":"Contribution of thyrotropin-releasing hormone to cerebellar long-term depression and motor learning.","volume":"12","author":"Watanave","year":"2018","journal-title":"Front. Cell Neurosci."},{"key":"B39","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.2113336119","article-title":"Protein kinase C\u03b3 in cerebellar Purkinje cells regulates Ca 2+-activated large-conductance K + channels and motor coordination.","volume":"119","author":"Watanave","year":"2022","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.45590","article-title":"TRPC3 is a major contributor to functional heterogeneity of cerebellar Purkinje cells.","volume":"8","author":"Wu","year":"2019","journal-title":"Elife"},{"key":"B41","doi-asserted-by":"crossref","DOI":"10.7554\/eLife.02536","article-title":"Cerebellar modules operate at different frequencies.","volume":"3","author":"Zhou","year":"2014","journal-title":"Elife"}],"container-title":["Frontiers in Cellular Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2024.1349878\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,16]],"date-time":"2024-02-16T04:34:43Z","timestamp":1708058083000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2024.1349878\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,16]]},"references-count":41,"alternative-id":["10.3389\/fncel.2024.1349878"],"URL":"https:\/\/doi.org\/10.3389\/fncel.2024.1349878","relation":{},"ISSN":["1662-5102"],"issn-type":[{"type":"electronic","value":"1662-5102"}],"subject":[],"published":{"date-parts":[[2024,2,16]]},"article-number":"1349878"}}