Abstract
It has previously been shown that the inhibition of L-type calcium channels (LTCCs) decreases alcohol consumption, although the contribution of the central LTCC subtypes Cav1.2 and Cav1.3 remains unknown. Here, we determined changes in Cav1.2 (Cacna1c) and Cav1.3 (Cacna1d) mRNA and protein expression in alcohol-dependent rats during protracted abstinence and naive controls using in situ hybridization and western blot analysis. Functional validation was obtained by electrophysiological recordings of calcium currents in dissociated hippocampal pyramidal neurons. We then measured alcohol self-administration and cue-induced reinstatement of alcohol seeking in dependent and nondependent rats after intracerebroventricular (i.c.v.) injection of the LTCC antagonist verapamil, as well as in mice with an inducible knockout (KO) of Cav1.2 in Ca2+/calmodulin-dependent protein kinase IIα (CaMKIIα)-expressing neurons. Our results show that Cacna1c mRNA concentration was increased in the amygdala and hippocampus of alcohol-dependent rats after 21 days of abstinence, with no changes in Cacna1d mRNA. This was associated with increased Cav1.2 protein concentration and L-type calcium current amplitudes. Further analysis of Cacna1c mRNA in the CA1, basolateral amygdala (BLA), and central amygdala (CeA) revealed a dynamic regulation over time during the development of alcohol dependence. The inhibition of central LTCCs via i.c.v. administration of verapamil prevented cue-induced reinstatement of alcohol seeking in alcohol-dependent rats. Further studies in conditional Cav1.2-KO mice showed a lack of dependence-induced increase of alcohol-seeking behavior. Together, our data indicate that central Cav1.2 channels, rather than Cav1.3, mediate alcohol-seeking behavior. This finding may be of interest for the development of new antirelapse medications.
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References
Baldelli P, Forni PE, Carbone E (2000). BDNF, NT-3 and NGF induce distinct new Ca2+ channel synthesis in developing hippocampal neurons. Eur J Neurosci 12: 4017–4032.
Berger SM, Bartsch D (2014). The role of L-type voltage-gated calcium channels Cav1.2 and Cav1.3 in normal and pathological brain function. Cell Tissue Res 357: 463–476.
Bernardi RE, Uhrig S, Spanagel R, Hansson AC (2014). Transcriptional regulation of L-type calcium channel subtypes Cav1.2 and Cav1.3 by nicotine and their potential role in nicotine sensitization. Nicotine Tob Res 16: 774–785.
Bjork K, Hansson AC, Sommer WH (2010). Genetic variation and brain gene expression in rodent models of alcoholism implications for medication development. Int Rev Neurobiol 91: 129–171.
Blackburn-Munro G, Brown CH, Neumann ID, Landgraf R, Russell JA (2000). Verapamil prevents withdrawal excitation of oxytocin neurones in morphine-dependent rats. Neuropharmacology 39: 1596–1607.
Carden WB, Alexander GM, Friedman DP, Daunais JB, Grant KA, Mu J et al (2006). Chronic ethanol drinking reduces native T-type calcium current in the thalamus of nonhuman primates. Brain Res 1089: 92–100.
Casamassima F, Hay AC, Benedetti A, Lattanzi L, Cassano GB, Perlis RH (2010). L-type calcium channels and psychiatric disorders: a brief review. Am J Med Genet B Neuropsychiatr Genet 153B: 1373–1390.
Clark NC, Nagano N, Kuenzi FM, Jarolimek W, Huber I, Walter D et al (2003). Neurological phenotype and synaptic function in mice lacking the CaV1.3 alpha subunit of neuronal L-type voltage-dependent Ca2+ channels. Neuroscience 120: 435–442.
Colombo G, Agabio R, Lobina C, Reali R, Melis F, Fadda F et al (1995). Effects of the calcium channel antagonist darodipine on ethanol withdrawal in rats. Alcohol Alcohol 30: 125–131.
De Beun R, Schneider R, Klein A, Lohmann A, De Vry J (1996a). Effects of nimodipine and other calcium channel antagonists in alcohol-preferring AA rats. Alcohol 13: 263–271.
De Beun R, Schneider R, Klein A, Lohmann A, Schreiber R, De Vry J (1996b). The calcium channel agonist BAY k 8644 reduces ethanol intake and preference in alcohol-preferring AA rats. Psychopharmacology (Berl) 127: 302–310.
Edwards S, Guerrero M, Ghoneim OM, Roberts E, Koob GF (2012). Evidence that vasopressin V1b receptors mediate the transition to excessive drinking in ethanol-dependent rats. Addict Biol 17: 76–85.
Erk S, Meyer-Lindenberg A, Schmierer P, Mohnke S, Grimm O, Garbusow M et al (2014). Hippocampal and frontolimbic function as intermediate phenotype for psychosis: evidence from healthy relatives and a common risk variant in CACNA1C. Biol Psychiatry 76: 466–475.
Fadda F, Garau B, Colombo G, Gessa GL (1992). Isradipine and other calcium channel antagonists attenuate ethanol consumption in ethanol-preferring rats. Alcohol Clin Exp Res 16: 449–452.
Gardell LR, Reid LD, Boedeker KL, Liakos TM, Hubbell CL (1997). Isradipine and naltrexone in combination with isradipine interact with a period of abstinence to reduce rats' intakes of an alcoholic beverage. Alcohol Clin Exp Res 21: 1592–1598.
Gardell LR, Reid ML, Cavallero CA, Burgess SE, Wallace RF, Hubbell CL et al (1999). Amlodipine, a calcium channel inhibitor, and cocaine and ethanol's reinforcing effects. Pharmacol Biochem Behav 64: 567–572.
Giordano TP, Satpute SS, Striessnig J, Kosofsky BE, Rajadhyaksha AM (2006). Up-regulation of dopamine D(2)L mRNA levels in the ventral tegmental area and dorsal striatum of amphetamine-sensitized C57BL/6 mice: role of Ca(v)1.3 L-type Ca(2+) channels. J Neurochemistry 99: 1197–1206.
Giordano TP, Tropea TF, Satpute SS, Sinnegger-Brauns MJ, Striessnig J, Kosofsky BE et al (2010). Molecular switch from L-type Ca v 1.3 to Ca v 1.2 Ca2+ channel signaling underlies long-term psychostimulant-induced behavioral and molecular plasticity. J Neurosci 30: 17051–17062.
Hansson AC, Cippitelli A, Sommer WH, Ciccocioppo R, Heilig M (2007). Region-specific down-regulation of Crhr1 gene expression in alcohol-preferring msP rats following ad lib access to alcohol. Addict Biol 12: 30–34.
Hansson AC, Cippitelli A, Sommer WH, Fedeli A, Bjork K, Soverchia L et al (2006). Variation at the rat Crhr1 locus and sensitivity to relapse into alcohol seeking induced by environmental stress. Proc Natl Acad Sci USA 103: 15236–15241.
Hansson AC, Nixon K, Rimondini R, Damadzic R, Sommer WH, Eskay R et al (2010). Long-term suppression of forebrain neurogenesis and loss of neuronal progenitor cells following prolonged alcohol dependence in rats. Int J Neuropsychopharmacol 13: 583–593.
Heilig M, Egli M, Crabbe JC, Becker HC (2010). Acute withdrawal, protracted abstinence and negative affect in alcoholism: are they linked? Addict Biol 15: 169–184.
Hell JW, Westenbroek RE, Warner C, Ahlijanian MK, Prystay W, Gilbert MM et al (1993). Identification and differential subcellular localization of the neuronal class C and class D L-type calcium channel alpha 1 subunits. J Cell Biol 123: 949–962.
Hermann D, Weber-Fahr W, Sartorius A, Hoerst M, Frischknecht U, Tunc-Skarka N et al (2012). Translational magnetic resonance spectroscopy reveals excessive central glutamate levels during alcohol withdrawal in humans and rats. Biol Psychiatry 71: 1015–1021.
Hirth N, Meinhardt MW, Noori HR, Hsalgado H, Torres-Ramirez O, Uhrig S et al (2016). Convergent evidence from alcohol dependent humans and rats for a hyperdopaminergic state in protracted abstinence. Proc Natl Acad Sci USA 113: 3024–3029.
Jacob SW, De La Torre JC (2009). Pharmacology of dimethyl sulfoxide in cardiac and CNS damage. Pharmacol Rep 61: 225–235.
Kelley AE (2004). Memory and addiction: shared neural circuitry and molecular mechanisms. Neuron 44: 161–179.
Koob GF, Le Moal M (2008). Addiction and the brain antireward system. Annu Rev Psychol 59: 29–53.
Ledoux J (2003). The emotional brain, fear, and the amygdala. Cell Mol Neurobiol 23: 727–738.
Liebmann L, Karst H, Sidiropoulou K, Van Gemert N, Meijer OC, Poirazi P et al (2008). Differential effects of corticosterone on the slow afterhyperpolarization in the basolateral amygdala and CA1 region: possible role of calcium channel subunits. J Neurophysiol 99: 958–968.
Lipscombe D (2002). L-type calcium channels: highs and new lows. Circ Res 90: 933–935.
Littleton JM, Little HJ, Whittington MA (1990). Effects of dihydropyridine calcium channel antagonists in ethanol withdrawal; doses required, stereospecificity and actions of Bay K 8644. Psychopharmacology (Berl) 100: 387–392.
Mahapatra S, Marcantoni A, Vandael DH, Striessnig J, Carbone E (2011). Are Ca(v)1.3 pacemaker channels in chromaffin cells? Possible bias from resting cell conditions and DHP blockers usage. Channels (Austin) 5: 219–224.
Mandyam CD (2013). The interplay between the hippocampus and amygdala in regulating aberrant hippocampal neurogenesis during protracted abstinence from alcohol dependence. Front Psychiatry 4: 61.
Marcantoni A, Vandael DH, Mahapatra S, Carabelli V, Sinnegger-Brauns MJ, Striessnig J et al (2010). Loss of Cav1.3 channels reveals the critical role of L-type and BK channel coupling in pacemaking mouse adrenal chromaffin cells. J Neurosci 30: 491–504.
Meinhardt MW, Hansson AC, Perreau-Lenz S, Bauder-Wenz C, Stahlin O, Heilig M et al (2013). Rescue of infralimbic mGluR2 deficit restores control over drug-seeking behavior in alcohol dependence. J Neurosci 33: 2794–2806.
Meinhardt MW, Sommer WH (2015). Postdependent state in rats as a model for medication development in alcoholism. Addict Biol 20: 1–21.
N'gouemo P, Akinfiresoye LR, Allard JS, Lovinger DM (2015). Alcohol withdrawal-induced seizure susceptibility is associated with an upregulation of CaV1.3 channels in the rat inferior colliculus. Int J Neuropsychopharmacol 18: pyu123.
N'gouemo P, Morad M (2003). Ethanol withdrawal seizure susceptibility is associated with upregulation of L- and P-type Ca2+ channel currents in rat inferior colliculus neurons. Neuropharmacology 45: 429–437.
Newton PM, Messing RO (2009). The N-type calcium channel is a novel target for treating alcohol use disorders. Channels (Austin) 3: 77–81.
Newton PM, Zeng L, Wang V, Connolly J, Wallace MJ, Kim C et al (2008). A blocker of N- and T-type voltage-gated calcium channels attenuates ethanol-induced intoxication, place preference, self-administration, and reinstatement. J Neurosci 28: 11712–11719.
Noori HR, Spanagel R, Hansson AC (2012). Neurocircuitry for modeling drug effects. Addict Biol 17: 827–864.
Nordskog BK, Hammarback JA, Godwin DW (2006). Diurnal gene expression patterns of T-type calcium channels and their modulation by ethanol. Neuroscience 141: 1365–1373.
Nunez-Santana FL, Oh MM, Antion MD, Lee A, Hell JW, Disterhoft JF (2014). Surface L-type Ca2+ channel expression levels are increased in aged hippocampus. Aging Cell 13: 111–120.
Otis JM, Fitzgerald MK, Mueller D (2014). Inhibition of hippocampal beta-adrenergic receptors impairs retrieval but not reconsolidation of cocaine-associated memory and prevents subsequent reinstatement. Neuropsychopharmacology 39: 303–310.
Pucilowski O, Krzascik P, Trzaskowska E, Kostowski W (1989). Different effect of diltiazem and nifedipine on some central actions of ethanol in the rat. Alcohol 6: 165–168.
Rashidy-Pour A, Vafaei AA, Taherian AA, Miladi-Gorji H, Sadeghi H, Fathollahi Y et al (2009). Verapamil enhances acute stress or glucocorticoid-induced deficits in retrieval of long-term memory in rats. Behav Brain Res 203: 76–80.
Rezvani AH, Janowsky DS (1990). Decreased alcohol consumption by verapamil in alcohol preferring rats. Prog Neuropsychopharmacol Biol Psychiatry 14: 623–631.
Rezvani AH, Mack CM, De Lacy PA, Janowsky DS (1990). Verapamil effects on physiological and behavioral responses to ethanol in the rat. Alcohol Alcohol 25: 51–58.
Rimondini R, Arlinde C, Sommer W, Heilig M (2002). Long-lasting increase in voluntary ethanol consumption and transcriptional regulation in the rat brain after intermittent exposure to alcohol. FASEB J 16: 27–35.
Schierberl K, Giordano T, Satpute S, Hao J, Kaur G, Hofmann F et al (2012). Cav 1.3L-type Ca (2+) channels mediate long-term adaptation in dopamine D2L-mediated GluA1 trafficking in the dorsal striatum following cocaine exposure. Channels (Austin) 6: 11–17.
Schierberl K, Hao J, Tropea TF, Ra S, Giordano TP, Xu Q et al (2011). Cav1.2L-type Ca(2)(+) channels mediate cocaine-induced GluA1 trafficking in the nucleus accumbens, a long-term adaptation dependent on ventral tegmental area Ca(v)1.3 channels. J Neurosci 31: 13562–13575.
Shi HS, Luo YX, Yin X, Wu HH, Xue G, Geng XH et al (2015). Reconsolidation of a cocaine associated memory requires DNA methyltransferase activity in the basolateral amygdala. Sci Rep 5: 13327.
Shibasaki M, Kurokawa K, Mizuno K, Ohkuma S (2011). Up-regulation of Ca(v)1.2 subunit via facilitating trafficking induced by Vps34 on morphine-induced place preference in mice. Eur J Pharmacol 651: 137–145.
Sommer WH, Rimondini R, Hansson AC, Hipskind PA, Gehlert DR, Barr CS et al (2008). Upregulation of voluntary alcohol intake, behavioral sensitivity to stress, and amygdala crhr1 expression following a history of dependence. Biol Psychiatry 63: 139–145.
Spanagel R (2009). Alcoholism: a systems approach from molecular physiology to addictive behavior. Physiol Rev 89: 649–705.
Spanagel R, Durstewitz D, Hansson A, Heinz A, Kiefer F, Kohr G et al (2013). A systems medicine research approach for studying alcohol addiction. Addict Biol 18: 883–896.
Strohmaier J, Amelang M, Hothorn LA, Witt SH, Nieratschker V, Gerhard D et al (2013). The psychiatric vulnerability gene CACNA1C and its sex-specific relationship with personality traits, resilience factors and depressive symptoms in the general population. Mol Psychiatry 18: 607–613.
Walker BM, Zorrilla EP, Koob GF (2011). Systemic kappa-opioid receptor antagonism by nor-binaltorphimine reduces dependence-induced excessive alcohol self-administration in rats. Addict Biol 16: 116–119.
Waltereit R, Mannhardt S, Nescholta S, Maser-Gluth C, Bartsch D (2008). Selective and protracted effect of nifedipine on fear memory extinction correlates with induced stress response. Learn Mem 15: 348–356.
World Health Organization (2014). Global status report on alcohol and health. www.who.int/substance_abuse/publications/alcohol_2014/en/.
Zhu B, Herbert J (1997). Calcium channels mediate angiotensin II-induced drinking behaviour and c-fos expression in the brain. Brain Res 778: 206–214.
Acknowledgements
We thank Elisabeth Röbel, Sabrina Koch, Merle Kochan, and Leonie Mehrke for their invaluable technical assistance. The study was supported by the German Federal Ministry of Education and Research (BMBF, e:Med program, FKZ: 01ZX1311A Spanagel et al, 2013, IntegraMent FKZ: 01ZX1314H, ERA-Net TRANSALC, FKZ 01EW1112), European Union's Horizon 2020 research and innovation program (668863, SyBil-AA), and Deutsche Forschungsgemeinschaft (DFG HA6102/1-1, DFG Center Grant (SFB1134)).
Author contributions
ACH, EC, RS, and WHS were responsible for study design and procured study funding. SU, REB, NH, and LB performed the mRNA analysis and behavioral experiments. ND and JMD performed the western blots. DV, AM, and EC conducted the electrophysiological experiments. KS and DB provided the Cav1.2fl/fl mice and demonstrated the Cav1.2 knockout. PG and MAV performed the behavioral characterization of the Cav1.2KO mice. AB was responsible for the operant behavior in mice. SU, AB, PG, MAV, REB, EC, JMD, and ACH analyzed data. RS, SU, EC, WHS, and ACH wrote the manuscript.
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Uhrig, S., Vandael, D., Marcantoni, A. et al. Differential Roles for L-Type Calcium Channel Subtypes in Alcohol Dependence. Neuropsychopharmacol 42, 1058–1069 (2017). https://doi.org/10.1038/npp.2016.266
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DOI: https://doi.org/10.1038/npp.2016.266
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