N-type calcium channel explained

N-type calcium channels also called Ca_v2.2 channels are voltage gated calcium channels that are localized primarily on the nerve terminals and dendrites as well as neuroendocrine cells.^[1] The calcium N-channel consists of several subunits: the primary subunit α1B and the auxiliary subunits α2δ and β. The α1B subunit forms the pore through which the calcium enters and helps to determine most of the channel's properties. These channels play an important role in the neurotransmission during development. In the adult nervous system, N-type calcium channels are critically involved in the release of neurotransmitters, and in pain pathways.^{[2] [3]} N-type calcium channels are the target of ziconotide, the drug prescribed to relieve intractable cancer pain. There are many known N-type calcium channel blockers that function to inhibit channel activity, although the most notable blockers are ω-conotoxins.^[4]

Structure

N-type calcium channels are categorized as high threshold-activated channels and seen in the Cav2 gene family. The structure of the N-type calcium channel is very similar to other voltage-dependent channels. The most important part of the channel is the actual pore that is formed by the α1B subunit. This pore is the location of the import of the extracellular ions. The α1B subunit has as many as 2000 amino acid residues within an amino acid sequence with the transmembrane structure with a pore. This is organized into 6 six segments(S1-S6). S1, S2, S3, S5, and S6 are hydrophobic while S4 serves as the voltage-sensor. In addition there is a membrane-associated loop in between S5 and S6. The activity of the pore is modulated by 4 subunits: an intracellular β-subunit, a transmembrane gamma subunit, and complex of alpha-2 and delta subunits.^[5]

In addition to the α1B subunit encoded by CACNA1B gene, the following auxiliary subunits are present in the N-type calcium channel:

• α2δ – encoded by either one of two genes CACNA2D1, CACNA2D2
• β – encoded by either one of four genes CACNB1, CACNB2, CACNB3, CACNB4

Function

N-type calcium channels are important in neurotransmitter release because they are localized at the synaptic terminals.^[6] In the peripheral nervous system, N-type channels are known to be involved in the release of many neurotransmitters like glutamate, GABA, acetylcholine, dopamine, and norepinephrine. When extracellular calcium flows into N-type calcium channels due to an action potential, it triggers the fusion of the secretory vesicles.

Studies on the cardiovascular system reveal when ω-Conotoxin is introduced, it causes the inhibition of norepinephrine, and this shows that only the N-type calcium channel, not the P/Q/L type calcium channels, are involved in the release of norepinephrine.^[7]

In the kidneys, blocking of N-type calcium channels reduce glomerular pressure through dilation of arterioles.^[8]

N-type calcium channels have been shown to play a part in the localization of neurite growth in the sympathetic nervous system and the skin and spinal cord. The neurite outgrowth was shown to be inhibited through an interaction between laminin and the 11th loop of the n-type calcium channel structure.^[9] It has been suggested that neurites outgrowth is inhibited by the influx of calcium through the growth cone, and this happens when the Cav2.2 channel comes in contact with laminin 2, and in response can induce a stretch activation of the N-type calcium channel.^[9]

Mutations

A rare gain-of-function mutation in CACNA1B gene encoding the α1B subunit of N-type calcium channel was suggested as the cause of several cases of a myoclonus-dystonia syndrome, although this suggestion has been disputed.^[3] Loss-of-function CACNA1B mutations were found to be present in progressive epilepsy-dyskinesia.^[10]

Clinical significance

The alteration of N-type calcium channels in therapeutic processes occurs in four major ways; through the blockage of N-type calcium channel peptides, interference of the flow of ions through the channel itself, activation of G-protein coupled signaling, and interference of the G-protein pathways.^[11] Studies have shown that the intrathecal injection of the calcium channel inhibitor ziconotide, to block the N-type calcium channels, have produced alleviation of intractable pain.^[12] Blockade of the N-type calcium channel is a potential therapeutic strategy for the treatment of alcoholism. Because prolonged alcohol exposure over time has been known to increase N-type channel function, experiments have shown that using N-type antagonists to decrease channel activity resulted in reduced voluntary consumption of alcohol in mice.^[13]

Blockers

In the pain pathway, N-type calcium channels serve to regulate pain signals sent from the peripheral nervous system to Central Nervous System. Although many N-type calcium channels blockers are known, most potent and selective belong to the family of conotoxins.^[14]

List of N-type Calcium channel blockers:

• ω-Conotoxins
• Cadmium
• Caroverine
• Cilnidipine
• Desipramine^[15]
• Gabapentin nonselectively inhibits N-type calcium channels by attaching to auxiliary α2δ subunit
• Levetiracetam
• Lamotrigine
• Nicardipine
• NP078585
• Piracetam^[16]
• Pregabalin nonselectively inhibits N-type calcium channels by attaching to auxiliary α2δ subunit
• TROX-1
• Ziconotide, a synthetic version of one of conotoxins

Further reading

• Calabrese B, Tabarean IV, Juranka P, Morris CE . Mechanosensitivity of N-type calcium channel currents . Biophysical Journal . 83 . 5 . 2560–74 . November 2002 . 12414690 . 1302342 . 10.1016/S0006-3495(02)75267-3 . 2002BpJ....83.2560C .
• Moskvina V, Craddock N, Holmans P, Nikolov I, Pahwa JS, Green E, Owen MJ, O'Donovan MC . Gene-wide analyses of genome-wide association data sets: evidence for multiple common risk alleles for schizophrenia and bipolar disorder and for overlap in genetic risk . Molecular Psychiatry . 14 . 3 . 252–60 . March 2009 . 19065143 . 10.1038/mp.2008.133 . 3970088 .
• Diane Lipscombe . Castiglioni AJ, Raingo J, Lipscombe D . Alternative splicing in the C-terminus of CaV2.2 controls expression and gating of N-type calcium channels . The Journal of Physiology . 576 . Pt 1 . 119–34 . October 2006 . 16857708 . 1995641 . 10.1113/jphysiol.2006.115030 .
• Catterall WA, Perez-Reyes E, Snutch TP, Striessnig J . International Union of Pharmacology. XLVIII. Nomenclature and structure-function relationships of voltage-gated calcium channels . Pharmacological Reviews . 57 . 4 . 411–25 . December 2005 . 16382099 . 10.1124/pr.57.4.5 . 10386627 .
• Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P, Mann M . Global, in vivo, and site-specific phosphorylation dynamics in signaling networks . Cell . 127 . 3 . 635–48 . November 2006 . 17081983 . 10.1016/j.cell.2006.09.026 . free .
• Stotz SC, Barr W, McRory JE, Chen L, Jarvis SE, Zamponi GW . Several structural domains contribute to the regulation of N-type calcium channel inactivation by the beta 3 subunit . The Journal of Biological Chemistry . 279 . 5 . 3793–800 . January 2004 . 14602720 . 10.1074/jbc.M308991200 . free .
• Maximov A, Bezprozvanny I . Synaptic targeting of N-type calcium channels in hippocampal neurons . The Journal of Neuroscience . 22 . 16 . 6939–52 . August 2002 . 12177192 . 3307533 . 10.1523/JNEUROSCI.22-16-06939.2002 .
• Peng S, Hajela RK, Atchison WD . Characteristics of block by Pb2+ of function of human neuronal L-, N-, and R-type Ca2+ channels transiently expressed in human embryonic kidney 293 cells . Molecular Pharmacology . 62 . 6 . 1418–30 . December 2002 . 12435810 . 10.1124/mol.62.6.1418 .
• Murakami M, Fleischmann B, De Felipe C, Freichel M, Trost C, Ludwig A, Wissenbach U, Schwegler H, Hofmann F, Hescheler J, Flockerzi V, Cavalié A . Pain perception in mice lacking the beta3 subunit of voltage-activated calcium channels . The Journal of Biological Chemistry . 277 . 43 . 40342–51 . October 2002 . 12161429 . 10.1074/jbc.M203425200 . free . 10261/310195 . free .
• Vitko I, Shcheglovitov A, Baumgart JP, Arias-Olguín II, Murbartián J, Arias JM, Perez-Reyes E . Orientation of the calcium channel beta relative to the alpha(1)2.2 subunit is critical for its regulation of channel activity . PLOS ONE . 3 . 10 . e3560 . 2008 . 18958281 . 2570331 . 10.1371/journal.pone.0003560 . Schwartz . 2008PLoSO...3.3560V . Arnold . free .
• Agler HL, Evans J, Tay LH, Anderson MJ, Colecraft HM, Yue DT . G protein-gated inhibitory module of N-type (ca(v)2.2) ca2+ channels . Neuron . 46 . 6 . 891–904 . June 2005 . 15953418 . 10.1016/j.neuron.2005.05.011 . free .

Notes and References

1. Catterall WA, Perez-Reyes E, Snutch TP,Striessnig J. Voltage-gated calcium channels (version 2019.4) in the IUPHAR/BPS Guide to Pharmacology Database . IUPHAR/BPS Guide to Pharmacology CITE . Edinburgh University Library . 2019 . 4 . 2019-09-16 . 2633-1020 . 10.2218/gtopdb/f80/2019.4 . free .
2. Web site: Voltage-dependent calcium channel, N-type, alpha-1 subunit. InterPro. EMBL-EBI.
3. Heyes S, Pratt WS, Rees E, Dahimene S, Ferron L, Owen MJ, Dolphin AC . Genetic disruption of voltage-gated calcium channels in psychiatric and neurological disorders . Prog Neurobiol . 134 . 36–54 . November 2015 . 26386135 . 4658333 . 10.1016/j.pneurobio.2015.09.002 .
4. Adams DJ, Berecki G . Mechanisms of conotoxin inhibition of N-type (Ca(v)2.2) calcium channels . Biochimica et Biophysica Acta (BBA) - Biomembranes . 1828 . 7 . 1619–28 . July 2013 . 23380425 . 10.1016/j.bbamem.2013.01.019 . free .
5. Web site: EMBL-EBI. InterPro. Voltage-dependent calcium channel, N-type, alpha-1 subunit (IPR005447) < InterPro < EMBL-EBI. www.ebi.ac.uk. en.
6. Weber AM, Wong FK, Tufford AR, Schlichter LC, Matveev V, Stanley EF . N-type Ca2+ channels carry the largest current: implications for nanodomains and transmitter release . Nature Neuroscience . 13 . 11 . 1348–50 . 2010 . 20953196 . 10.1038/nn.2657 . 205433301 .
7. Molderings GJ, Likungu J, Göthert M . N-Type calcium channels control sympathetic neurotransmission in human heart atrium . Circulation . 101 . 4 . 403–7 . February 2000 . 10653832 . 10.1161/01.cir.101.4.403. free .
8. Hayashi K, Wakino S, Sugano N, Ozawa Y, Homma K, Saruta T . Ca2+ channel subtypes and pharmacology in the kidney . Circulation Research . 100 . 3 . 342–53 . February 2007 . 17307972 . 10.1161/01.RES.0000256155.31133.49 . free .
9. Weiss N . The N-type voltage-gated calcium channel: when a neuron reads a map . The Journal of Neuroscience . 28 . 22 . 5621–2 . May 2008 . 18509022 . 10.1523/JNEUROSCI.1538-08.2008 . 6670785 .
10. Gorman KM, Meyer E, Grozeva D, Spinelli E, McTague A, Sanchis-Juan A, Carss KJ, Bryant E, Reich A, Schneider AL, Pressler RM, Simpson MA, Debelle GD, Wassmer E, Morton J, Sieciechowicz D, Jan-Kamsteeg E, Paciorkowski AR, King MD, Cross JH, Poduri A, Mefford HC, Scheffer IE, Haack TB, McCullagh G, Millichap JJ, Carvill GL, Clayton-Smith J, Maher ER, Raymond FL, Kurian MA . 6. Bi-allelic Loss-of-Function CACNA1B Mutations in Progressive Epilepsy-Dyskinesia . Am J Hum Genet . 104 . 5 . 948–956 . May 2019 . 30982612 . 6507039 . 10.1016/j.ajhg.2019.03.005 .
11. Zamponi GW, Striessnig J, Koschak A, Dolphin AC . The Physiology, Pathology, and Pharmacology of Voltage-Gated Calcium Channels and Their Future Therapeutic Potential . Pharmacological Reviews . 67 . 4 . 821–70 . October 2015 . 26362469 . 10.1124/pr.114.009654 . Zamponi . 4630564.
12. Dray A, Read SJ . Arthritis and pain. Future targets to control osteoarthritis pain . Arthritis Research & Therapy . 9 . 3 . 212 . May 2007 . 17561993 . 2206352 . 10.1186/ar2178 . free .
13. Newton PM, Zeng L, Wang V, Connolly J, Wallace MJ, Kim C, Shin HS, Belardetti F, Snutch TP, Messing RO . A blocker of N- and T-type voltage-gated calcium channels attenuates ethanol-induced intoxication, place preference, self-administration, and reinstatement . The Journal of Neuroscience . 28 . 45 . 11712–9 . November 2008 . 18987207 . 10.1523/JNEUROSCI.3621-08.2008 . Newton . 3045811.
14. 2013-07-01. Mechanisms of conotoxin inhibition of N-type (Cav2.2) calcium channels. Biochimica et Biophysica Acta (BBA) - Biomembranes. en. 1828. 7. 1619–1628. 10.1016/j.bbamem.2013.01.019. 23380425. 0005-2736. Adams. David J.. Berecki. Géza. free.
15. Benjamin ER, Pruthi F, Olanrewaju S, Shan S, Hanway D, Liu X, Cerne R, Lavery D, Valenzano KJ, Woodward RM, Ilyin VI . Pharmacological characterization of recombinant N-type calcium channel (Cav2.2) mediated calcium mobilization using FLIPR . Biochem Pharmacol . 72 . 6 . 770–82 . September 2006 . 16844100 . 10.1016/j.bcp.2006.06.003 .
16. Bravo-Martínez J, Arenas I, Vivas O, Rebolledo-Antúnez S, Vázquez-García M, Larrazolo A, García DE . A novel CaV2.2 channel inhibition by piracetam in peripheral and central neurons . Exp Biol Med (Maywood) . 237 . 10 . 1209–18 . October 2012 . 23045722 . 10.1258/ebm.2012.012128 . 25909697 .