{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T15:47:27Z","timestamp":1764604047827,"version":"build-2065373602"},"reference-count":58,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,1,7]],"date-time":"2024-01-07T00:00:00Z","timestamp":1704585600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"IPSEN bio innovation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Botulinum neurotoxin A (BoNT\/A) is a potent neurotoxin that silences cholinergic neurotransmission through the cleavage of the synaptic protein SNAP-25. Previous studies have shown that, in addition to its paralytic effects, BoNT\/A can inhibit sensory nerve activity. The aim of this study was to identify how BoNT\/A inhibits afferent signalling from the bladder. To investigate the role of SNAP-25 cleavage in the previously reported BoNT\/A-dependent inhibition of sensory signalling, we developed a recombinant form of BoNT\/A with an inactive light chain, rBoNT\/A (0), unable to paralyse muscle. We also developed recombinant light chain (LC)-domain-only proteins to better understand the entry mechanisms, as the heavy chain (HC) of the protein is responsible for the internalisation of the light chain. We found that, despite a lack of catalytic activity, rBoNT\/A (0) potently inhibited the afferent responses to bladder distension to a greater degree than catalytically active rBoNT\/A. This was also clear from the testing of the LC-only proteins, as the inactive rLC\/A (0) protein inhibited afferent responses significantly more than the active rLC\/A protein. Immunohistochemistry for cleaved SNAP-25 was negative, and purinergic and nitrergic antagonists partially and totally reversed the sensory inhibition, respectively. These data suggest that the BoNT\/A inhibition of sensory nerve activity in this assay is not due to the classical well-characterised \u2018double-receptor\u2019 mechanism of BoNT\/A, is independent of SNAP25 cleavage and involves nitrergic and purinergic signalling mechanisms.<\/jats:p>","DOI":"10.3390\/toxins16010030","type":"journal-article","created":{"date-parts":[[2024,1,8]],"date-time":"2024-01-08T05:21:38Z","timestamp":1704691298000},"page":"30","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["A Novel Catalytically Inactive Construct of Botulinum Neurotoxin A (BoNT\/A) Directly Inhibits Visceral Sensory Signalling"],"prefix":"10.3390","volume":"16","author":[{"given":"Hodan","family":"Ibrahim","sequence":"first","affiliation":[{"name":"School of Pharmacy and Biomedical Sciences, University of Central Lancashire, Preston Campus, Preston PR1 2HE, UK"},{"name":"Ipsen, Abingdon OX14 4RY, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7506-4859","authenticated-orcid":false,"given":"Kevin","family":"Retailleau","sequence":"additional","affiliation":[{"name":"Ipsen, 5 av Canada, 91940 Les Ulis, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fraser","family":"Hornby","sequence":"additional","affiliation":[{"name":"Ipsen, Abingdon OX14 4RY, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2512-5593","authenticated-orcid":false,"given":"Jacquie","family":"Maignel","sequence":"additional","affiliation":[{"name":"Ipsen, 5 av Canada, 91940 Les Ulis, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3249-7851","authenticated-orcid":false,"given":"Matthew","family":"Beard","sequence":"additional","affiliation":[{"name":"Ipsen, Abingdon OX14 4RY, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9026-8200","authenticated-orcid":false,"given":"Donna Marie","family":"Daly","sequence":"additional","affiliation":[{"name":"School of Pharmacy and Biomedical Sciences, University of Central Lancashire, Preston Campus, Preston PR1 2HE, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1016\/j.coph.2004.12.006","article-title":"Botulinal neurotoxins: Revival of an old killer","volume":"5","author":"Montecucco","year":"2005","journal-title":"Curr. 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