{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T05:45:22Z","timestamp":1792043122141,"version":"4.4.0"},"reference-count":43,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,6,17]],"date-time":"2024-06-17T00:00:00Z","timestamp":1718582400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,6,17]],"date-time":"2024-06-17T00:00:00Z","timestamp":1718582400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001794","name":"The University of Queensland","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100001794","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Cell. Mol. Life Sci."],"published-print":{"date-parts":[[2024,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Acid-sensing ion channels (ASICs) are trimeric proton-gated cation channels that play a role in neurotransmission and pain sensation. The snake venom-derived peptides, mambalgins, exhibit potent analgesic effects in rodents by inhibiting central ASIC1a and peripheral ASIC1b. Despite their distinct species- and subtype-dependent pharmacology, previous structure-function studies have focussed on the mambalgin interaction with ASIC1a. Currently, the specific channel residues responsible for this pharmacological profile, and the mambalgin pharmacophore at ASIC1b remain unknown. Here we identify non-conserved residues at the ASIC1 subunit interface that drive differences in the mambalgin pharmacology from rat ASIC1a to ASIC1b, some of which likely do not make peptide binding interactions. Additionally, an amino acid variation below the core binding site explains potency differences between rat and human ASIC1. Two regions within the palm domain, which contribute to subtype-dependent effects for mambalgins, play key roles in ASIC gating, consistent with subtype-specific differences in the peptides mechanism. Lastly, there is a shared primary mambalgin pharmacophore for ASIC1a and ASIC1b activity, with certain peripheral peptide residues showing variant-specific significance for potency. Through our broad mutagenesis studies across various species and subtype variants, we gain a more comprehensive understanding of the pharmacophore and the intricate molecular interactions that underlie ligand specificity. These insights pave the way for the development of more potent and targeted peptide analogues required to advance our understating of human ASIC1 function and its role in disease.<\/jats:p>","DOI":"10.1007\/s00018-024-05276-2","type":"journal-article","created":{"date-parts":[[2024,6,16]],"date-time":"2024-06-16T23:10:15Z","timestamp":1718579415000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Revealing molecular determinants governing mambalgin-3 pharmacology at acid-sensing ion channel 1 variants"],"prefix":"10.1007","volume":"81","author":[{"given":"Ben","family":"Cristofori-Armstrong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Budusan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer J.","family":"Smith","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steve","family":"Reynaud","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kerstin","family":"Voll","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ir\u00e8ne R.","family":"Chassagnon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Durek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3616-0279","authenticated-orcid":false,"given":"Lachlan D.","family":"Rash","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,6,17]]},"reference":[{"issue":"15","key":"5276_CR1","doi-asserted-by":"publisher","first-page":"1750","DOI":"10.2174\/138161209788186263","volume":"15","author":"GR Dub\u00e9","year":"2009","unstructured":"Dub\u00e9 GR, Elagoz A, Mangat H (2009) Acid sensing ion channels and acid nociception. Curr Pharm Design 15(15):1750\u20131766","journal-title":"Curr Pharm Design"},{"issue":"1","key":"5276_CR2","doi-asserted-by":"publisher","first-page":"150","DOI":"10.1016\/0006-8993(81)90446-7","volume":"214","author":"OA Krishtal","year":"1981","unstructured":"Krishtal OA, Pidoplichko VI (1981) Receptor for protons in the membrane of sensory neurons. Brain Res 214(1):150\u2013154","journal-title":"Brain Res"},{"issue":"7373","key":"5276_CR3","doi-asserted-by":"publisher","first-page":"410","DOI":"10.1038\/nature10607","volume":"479","author":"CJ Bohlen","year":"2011","unstructured":"Bohlen CJ et al (2011) A heteromeric Texas coral snake toxin targets acid-sensing ion channels to produce pain. Nature 479(7373):410\u2013414","journal-title":"Nature"},{"key":"5276_CR4","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1016\/j.neuropharm.2017.04.042","volume":"127","author":"B Cristofori-Armstrong","year":"2017","unstructured":"Cristofori-Armstrong B, Rash LD (2017) Acid-sensing ion channel (ASIC) structure and function: insights from spider, snake and sea anemone venoms. Neuropharmacology 127:173\u2013184","journal-title":"Neuropharmacology"},{"issue":"22","key":"5276_CR5","doi-asserted-by":"publisher","first-page":"3047","DOI":"10.1038\/emboj.2008.213","volume":"27","author":"E Deval","year":"2008","unstructured":"Deval E et al (2008) ASIC3, a sensor of acidic and primary inflammatory pain. EMBO J 27(22):3047\u20133055","journal-title":"EMBO J"},{"key":"5276_CR6","doi-asserted-by":"publisher","first-page":"982689","DOI":"10.3389\/fnmol.2022.982689","volume":"15","author":"J Ridley","year":"2022","unstructured":"Ridley J et al (2022) Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms. Front Mol Neurosci 15:982689","journal-title":"Front Mol Neurosci"},{"issue":"10","key":"5276_CR7","doi-asserted-by":"publisher","first-page":"709","DOI":"10.3390\/toxins14100709","volume":"14","author":"C Verkest","year":"2022","unstructured":"Verkest C et al (2022) Mechanisms of action of the peptide toxins Targeting Human and Rodent Acid-sensing Ion channels and relevance to their in vivo Analgesic effects. Toxins 14(10):709","journal-title":"Toxins"},{"issue":"7421","key":"5276_CR8","doi-asserted-by":"publisher","first-page":"552","DOI":"10.1038\/nature11494","volume":"490","author":"S Diochot","year":"2012","unstructured":"Diochot S et al (2012) Black mamba venom peptides target acid-sensing ion channels to abolish pain. Nature 490(7421):552\u2013555","journal-title":"Nature"},{"key":"5276_CR9","doi-asserted-by":"crossref","unstructured":"Chang C-T et al (2019) Involvement of acid-sensing Ion Channel 1b in the development of Acid-Induced Chronic muscle Pain. Frontiers in Neuroscience, p 13","DOI":"10.3389\/fnins.2019.01247"},{"issue":"3","key":"5276_CR10","doi-asserted-by":"publisher","first-page":"552","DOI":"10.1097\/j.pain.0000000000000397","volume":"157","author":"S Diochot","year":"2016","unstructured":"Diochot S et al (2016) Analgesic effects of mambalgin peptide inhibitors of acid-sensing ion channels in inflammatory and neuropathic pain. Pain 157(3):552\u2013559","journal-title":"Pain"},{"issue":"21","key":"5276_CR11","doi-asserted-by":"publisher","first-page":"4154","DOI":"10.1111\/bph.14462","volume":"175","author":"C Verkest","year":"2018","unstructured":"Verkest C et al (2018) Effects of systemic inhibitors of acid-sensing ion channels 1 (ASIC1) against acute and chronic mechanical allodynia in a rodent model of migraine. Br J Pharmacol 175(21):4154\u20134166","journal-title":"Br J Pharmacol"},{"key":"5276_CR12","doi-asserted-by":"crossref","unstructured":"Cristofori-Armstrong B, Budusan E, Rash LD (2021) Mambalgin-3 potentiates human acid-sensing ion channel 1b under mild to moderate acidosis: implications as an analgesic lead. Proc Natl Acad Sci 118(8)","DOI":"10.1073\/pnas.2021581118"},{"issue":"6","key":"5276_CR13","doi-asserted-by":"publisher","first-page":"2616","DOI":"10.1074\/jbc.M115.702373","volume":"291","author":"G Mourier","year":"2016","unstructured":"Mourier G et al (2016) Mambalgin-1 Pain-relieving peptide, stepwise solid-phase synthesis, Crystal structure, and functional domain for acid-sensing Ion Channel 1a inhibition. J Biol Chem 291(6):2616\u20132629","journal-title":"J Biol Chem"},{"key":"5276_CR14","doi-asserted-by":"publisher","first-page":"108453","DOI":"10.1016\/j.neuropharm.2021.108453","volume":"185","author":"M Salinas","year":"2021","unstructured":"Salinas M et al (2021) Mambalgin-1 pain-relieving peptide locks the hinge between alpha4 and alpha5 helices to inhibit rat acid-sensing ion channel 1a. Neuropharmacology 185:108453","journal-title":"Neuropharmacology"},{"key":"5276_CR15","doi-asserted-by":"crossref","unstructured":"Sun D et al (2020) Structural insights into human acid-sensing ion channel 1a inhibition by snake toxin mambalgin1. eLife 9","DOI":"10.7554\/eLife.57096"},{"issue":"4","key":"5276_CR16","doi-asserted-by":"publisher","first-page":"1017","DOI":"10.1002\/anie.201308898","volume":"53","author":"CI Schroeder","year":"2014","unstructured":"Schroeder CI et al (2014) Chemical synthesis, 3D structure, and ASIC binding site of the Toxin Mambalgin-2. Angew Chem Int Ed 53(4):1017\u20131020","journal-title":"Angew Chem Int Ed"},{"key":"5276_CR17","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1007\/978-1-4939-6887-9_10","volume":"1586","author":"NJ Saez","year":"2017","unstructured":"Saez NJ, Cristofori-Armstrong B, Anangi R, King GF (2017) A strategy for production of correctly folded disulfide-rich peptides in the periplasm of E. Coli. Methods Mol Biol 1586:155\u2013180","journal-title":"Methods Mol Biol"},{"key":"5276_CR18","doi-asserted-by":"publisher","first-page":"14763","DOI":"10.1038\/srep14763","volume":"5","author":"B Cristofori-Armstrong","year":"2015","unstructured":"Cristofori-Armstrong B et al (2015) Xenopus borealis as an alternative source of oocytes for biophysical and pharmacological studies of neuronal ion channels. Sci Rep 5:14763","journal-title":"Sci Rep"},{"issue":"46","key":"5276_CR19","doi-asserted-by":"publisher","first-page":"45240","DOI":"10.1074\/jbc.M304441200","volume":"278","author":"T Coric","year":"2003","unstructured":"Coric T, Zhang P, Todorovic N, Canessa CM (2003) The extracellular domain determines the kinetics of desensitization in acid-sensitive ion channel 1. J Biol Chem 278(46):45240\u201345247","journal-title":"J Biol Chem"},{"issue":"44","key":"5276_CR20","doi-asserted-by":"publisher","first-page":"41597","DOI":"10.1074\/jbc.M205877200","volume":"277","author":"E Babini","year":"2002","unstructured":"Babini E, Paukert M, Geisler HS, Grunder S (2002) Alternative splicing and interaction with di- and polyvalent cations control the dynamic range of acid-sensing ion channel 1 (ASIC1). J Biol Chem 277(44):41597\u201341603","journal-title":"J Biol Chem"},{"issue":"3","key":"5276_CR21","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1085\/jgp.200509409","volume":"127","author":"X Chen","year":"2006","unstructured":"Chen X, Kalbacher H, Gr\u00fcnder S (2006) Interaction of Acid-sensing Ion Channel (ASIC) 1 with the Tarantula Toxin Psalmotoxin 1 is State Dependent. J Gen Physiol 127(3):267\u2013276","journal-title":"J Gen Physiol"},{"issue":"19","key":"5276_CR22","doi-asserted-by":"publisher","first-page":"13363","DOI":"10.1074\/jbc.M114.561076","volume":"289","author":"M Salinas","year":"2014","unstructured":"Salinas M et al (2014) Binding site and inhibitory mechanism of the mambalgin-2 pain-relieving peptide on acid-sensing ion channel 1a. J Biol Chem 289(19):13363\u201313373","journal-title":"J Biol Chem"},{"issue":"7","key":"5276_CR23","doi-asserted-by":"publisher","first-page":"1837","DOI":"10.3390\/cancers12071837","volume":"12","author":"M Bychkov","year":"2020","unstructured":"Bychkov M et al (2020) Mambalgin-2 induces cell cycle arrest and apoptosis in glioma cells via Interaction with ASIC1a. Cancers 12(7):1837","journal-title":"Cancers"},{"issue":"10","key":"5276_CR24","doi-asserted-by":"publisher","first-page":"1324","DOI":"10.3390\/biomedicines9101324","volume":"9","author":"ML Bychkov","year":"2021","unstructured":"Bychkov ML et al (2021) Mambalgin-2 inhibits Growth, Migration, and Invasion of metastatic melanoma cells by targeting the channels containing an ASIC1a subunit whose Up-Regulation correlates with poor survival prognosis. Biomedicines 9(10):1324","journal-title":"Biomedicines"},{"issue":"10","key":"5276_CR25","doi-asserted-by":"publisher","first-page":"612","DOI":"10.3390\/toxins15100612","volume":"15","author":"EN Lyukmanova","year":"2023","unstructured":"Lyukmanova EN et al (2023) Molecular basis for Mambalgin-2 Interaction with Heterotrimeric \u03b1-ENaC\/ASIC1a\/\u03b3-ENaC channels in Cancer cells. Toxins 15(10):612","journal-title":"Toxins"},{"key":"5276_CR26","doi-asserted-by":"publisher","first-page":"904742","DOI":"10.3389\/fonc.2022.904742","volume":"12","author":"AV Sudarikova","year":"2022","unstructured":"Sudarikova AV et al (2022) Mambalgin-2 inhibits Lung Adenocarcinoma Growth and Migration by Selective Interaction with ASIC1\/\u03b1-ENaC\/\u03b3-ENaC Heterotrimer. Front Oncol 12:904742","journal-title":"Front Oncol"},{"key":"5276_CR27","doi-asserted-by":"publisher","first-page":"429","DOI":"10.1016\/j.neuropharm.2017.08.004","volume":"125","author":"T Besson","year":"2017","unstructured":"Besson T, Lingueglia E, Salinas M (2017) Pharmacological modulation of acid-sensing Ion channels 1a and 3 by amiloride and 2-guanidine-4-methylquinazoline (GMQ). Neuropharmacology 125:429\u2013440","journal-title":"Neuropharmacology"},{"issue":"5","key":"5276_CR28","doi-asserted-by":"publisher","first-page":"320","DOI":"10.1002\/psc.2868","volume":"22","author":"H Lan","year":"2016","unstructured":"Lan H et al (2016) Total synthesis of mambalgin-1\/2\/3 by two-segment hydrazide-based native chemical ligation. J Pept Sci 22(5):320\u2013326","journal-title":"J Pept Sci"},{"issue":"44","key":"5276_CR29","doi-asserted-by":"publisher","first-page":"5837","DOI":"10.1039\/C4CC00779D","volume":"50","author":"M Pan","year":"2014","unstructured":"Pan M et al (2014) One-pot hydrazide-based native chemical ligation for efficient chemical synthesis and structure determination of toxin Mambalgin-1. Chem Commun (Camb) 50(44):5837\u20135839","journal-title":"Chem Commun (Camb)"},{"key":"5276_CR30","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1038\/s41421-018-0026-1","volume":"4","author":"D Sun","year":"2018","unstructured":"Sun D et al (2018) Cryo-EM structure of the ASIC1a-mambalgin-1 complex reveals that the peptide toxin mambalgin-1 inhibits acid-sensing ion channels through an unusual allosteric effect. Cell Discov 4:27","journal-title":"Cell Discov"},{"issue":"38","key":"5276_CR31","doi-asserted-by":"publisher","first-page":"8153","DOI":"10.1039\/C5CC01418B","volume":"51","author":"M Wen","year":"2015","unstructured":"Wen M et al (2015) Site-specific fluorescence spectrum detection and characterization of hASIC1a channels upon toxin mambalgin-1 binding in live mammalian cells. Chem Commun (Camb) 51(38):8153\u20138156","journal-title":"Chem Commun (Camb)"},{"issue":"41","key":"5276_CR32","doi-asserted-by":"publisher","first-page":"31285","DOI":"10.1074\/jbc.M110.160382","volume":"285","author":"T Li","year":"2010","unstructured":"Li T, Yang Y, Canessa CM (2010) Asn415 in the beta11-beta12 linker decreases proton-dependent desensitization of ASIC1. J Biol Chem 285(41):31285\u201331291","journal-title":"J Biol Chem"},{"issue":"4","key":"5276_CR33","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1038\/s41573-020-00135-8","volume":"20","author":"M Muttenthaler","year":"2021","unstructured":"Muttenthaler M, King GF, Adams DJ, Alewood PF (2021) Trends in peptide drug discovery. Nat Rev Drug Discov 20(4):309\u2013325","journal-title":"Nat Rev Drug Discov"},{"issue":"27","key":"5276_CR34","doi-asserted-by":"publisher","first-page":"24374","DOI":"10.1074\/jbc.M111.230797","volume":"286","author":"A Springauf","year":"2011","unstructured":"Springauf A, Bresenitz P, Gr\u00fcnder S (2011) The interaction between two extracellular linker regions controls sustained opening of acid-sensing ion channel 1. J Biol Chem 286(27):24374\u201324384","journal-title":"J Biol Chem"},{"key":"5276_CR35","doi-asserted-by":"publisher","first-page":"e51111","DOI":"10.7554\/eLife.51111","volume":"9","author":"ML Rook","year":"2020","unstructured":"Rook ML et al (2020) \u03b211\u201312 linker isomerization governs acid-sensing ion channel desensitization and recovery. eLife 9:e51111","journal-title":"eLife"},{"key":"5276_CR36","doi-asserted-by":"publisher","first-page":"e45851","DOI":"10.7554\/eLife.45851","volume":"8","author":"Y Wu","year":"2019","unstructured":"Wu Y, Chen Z, Canessa CM (2019) A valve-like mechanism controls desensitization of functional mammalian isoforms of acid-sensing ion channels. eLife 8:e45851","journal-title":"eLife"},{"issue":"2","key":"5276_CR37","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1113\/JP278707","volume":"599","author":"ML Rook","year":"2021","unstructured":"Rook ML, Musgaard M, MacLean DM (2021) Coupling structure with function in acid-sensing ion channels: challenges in pursuit of proton sensors. J Physiol 599(2):417\u2013430","journal-title":"J Physiol"},{"key":"5276_CR38","doi-asserted-by":"publisher","first-page":"58","DOI":"10.3389\/fmolb.2020.00058","volume":"7","author":"O Bignucolo","year":"2020","unstructured":"Bignucolo O et al (2020) Structural and functional analysis of Gly212 mutants reveals the importance of Intersubunit interactions in ASIC1a Channel function. Front Mol Biosci 7:58","journal-title":"Front Mol Biosci"},{"issue":"7696","key":"5276_CR39","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1038\/nature25782","volume":"555","author":"N Yoder","year":"2018","unstructured":"Yoder N, Yoshioka C, Gouaux E (2018) Gating mechanisms of acid-sensing ion channels. Nature 555(7696):397\u2013401","journal-title":"Nature"},{"issue":"29","key":"5276_CR40","doi-asserted-by":"publisher","first-page":"17956","DOI":"10.1074\/jbc.M115.641688","volume":"290","author":"K Gwiazda","year":"2015","unstructured":"Gwiazda K, Bonifacio G, Vullo S, Kellenberger S (2015) Extracellular subunit interactions control transitions between Functional States of Acid-sensing Ion Channel 1a. J Biol Chem 290(29):17956\u201317966","journal-title":"J Biol Chem"},{"issue":"21","key":"5276_CR41","doi-asserted-by":"publisher","first-page":"16315","DOI":"10.1074\/jbc.M109.092015","volume":"285","author":"LA Liechti","year":"2010","unstructured":"Liechti LA et al (2010) A combined computational and functional approach identifies new residues involved in pH-dependent gating of ASIC1a. J Biol Chem 285(21):16315\u201316329","journal-title":"J Biol Chem"},{"key":"5276_CR42","doi-asserted-by":"publisher","first-page":"381","DOI":"10.1016\/j.bcp.2019.03.004","volume":"163","author":"B Cristofori-Armstrong","year":"2019","unstructured":"Cristofori-Armstrong B et al (2019) The modulation of acid-sensing ion channel 1 by PcTx1 is pH-, subtype- and species-dependent: importance of interactions at the channel subunit interface and potential for engineering selective analogues. Biochem Pharmacol 163:381\u2013390","journal-title":"Biochem Pharmacol"},{"key":"5276_CR43","doi-asserted-by":"crossref","unstructured":"Heusser SA, Borg CB, Colding JM, Pless SA (2022) Conformational decoupling in acid-sensing ion channels uncovers mechanism and stoichiometry of PcTx1-mediated inhibition. eLife 11:e73384","DOI":"10.7554\/eLife.73384"}],"container-title":["Cellular and Molecular Life Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00018-024-05276-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s00018-024-05276-2\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00018-024-05276-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,16]],"date-time":"2024-06-16T23:15:09Z","timestamp":1718579709000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s00018-024-05276-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,17]]},"references-count":43,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2024,12]]}},"alternative-id":["5276"],"URL":"https:\/\/doi.org\/10.1007\/s00018-024-05276-2","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-4539352\/v1","asserted-by":"object"}]},"ISSN":["1420-682X","1420-9071"],"issn-type":[{"value":"1420-682X","type":"print"},{"value":"1420-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6,17]]},"assertion":[{"value":"22 January 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 April 2024","order":2,"name":"revised","label":"Revised","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 May 2024","order":3,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"17 June 2024","order":4,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Animal care and procedures were carried out in strict accordance with the recommendations in the Australian code of practice for the care and use of animals for scientific purposes (8th Edition, 2013). The Anatomical Biosciences group of the Animal Ethics Committee at The University of Queensland approved surgery protocols (Approval numbers: 2018\/AE000421 and 2019\/AE000097). Animal surgery was performed under anaesthesia (frogs were bathed in ice-cold water containing 1.3\u00a0mg\/ml of buffered MS-222, ethyl 3-aminobenzoate methane sulfonate), and all efforts were made to minimise animal suffering. Three months was the minimum time between surgeries on the same frog, and after the final surgery (maximum of six surgeries per animal) frogs were euthanised by decapitation under ice and MS-222 anaesthesia.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}},{"value":"Author LDR is a co-inventor on a patent (US 10,485,847) for the use of the ASIC inhibitory peptide Hi1a as a neuroprotective agent. All other authors have no relevant financial or non-financial interests to disclose.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"<!--Emphasis Type='Bold' removed-->Competing interests"}}],"article-number":"266"}}