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The \u03b17 nAChR has high Ca2+ permeability and can be quickly activated and desensitized, and is closely related to Alzheimer\u2019s disease (AD), epilepsy, schizophrenia, lung cancer, Parkinson\u2019s disease (PD), inflammation, and other diseases. \u03b1-conotoxins from marine cone snail venom are typically short, disulfide-rich neuropeptides targeting nAChRs and can distinguish various subtypes, providing vital pharmacological tools for the functional research of nAChRs. [Q1G, \u0394R14]Lv\u0399B is a rat \u03b17 nAChRs selective antagonist, modified from \u03b1-conotoxin Lv\u0399B. In this study, we utilized three types of fluorescein after N-Hydroxy succinimide (NHS) activation treatment: 6-TAMRA-SE, Cy3 NHS, and BODIPY-FL NHS, labeling the N-Terminal of [Q1G, \u0394R14]Lv\u0399B under weak alkaline conditions, obtaining three fluorescent analogs: LvIB-R, LvIB-C, and LvIB-B, respectively. The potency of [Q1G, \u0394R14]Lv\u0399B fluorescent analogs was evaluated at rat \u03b17 nAChRs expressed in Xenopus laevis oocytes. Using a two-electrode voltage clamp (TEVC), the half-maximal inhibitory concentration (IC50) values of LvIB-R, LvIB-C, and LvIB-B were 643.3 nM, 298.0 nM, and 186.9 nM, respectively. The stability of cerebrospinal fluid analysis showed that after incubation for 12 h, the retention rates of the three fluorescent analogs were 52.2%, 22.1%, and 0%, respectively. [Q1G, \u0394R14]Lv\u0399B fluorescent analogs were applied to explore the distribution of \u03b17 nAChRs in the hippocampus and striatum of rat brain tissue and it was found that Cy3- and BODIPY FL-labeled [Q1G, \u0394R14]Lv\u0399B exhibited better imaging characteristics than 6-TAMARA-. It was also found that \u03b17 nAChRs are widely distributed in the cerebral cortex and cerebellar lobules. Taking into account potency, imaging, and stability, [Q1G, \u0394R14]Lv\u0399B -BODIPY FL is an ideal pharmacological tool to investigate the tissue distribution and function of \u03b17 nAChRs. Our findings not only provide a foundation for the development of conotoxins as visual pharmacological probes, but also demonstrate the distribution of \u03b17 nAChRs in the rat brain.<\/jats:p>","DOI":"10.3390\/md22050200","type":"journal-article","created":{"date-parts":[[2024,4,29]],"date-time":"2024-04-29T06:13:22Z","timestamp":1714371202000},"page":"200","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Fluorescent \u03b1-Conotoxin [Q1G, \u0394R14]LvIB Identifies the Distribution of \u03b17 Nicotinic Acetylcholine Receptor in the Rat Brain"],"prefix":"10.3390","volume":"22","author":[{"given":"Hongyu","family":"Shan","sequence":"first","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nan","family":"Wang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinyu","family":"Gao","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zihan","family":"Wang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4936-8637","authenticated-orcid":false,"given":"Jinpeng","family":"Yu","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dongting","family":"Zhangsun","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"},{"name":"Key Laboratory of Tropical Biological Resources of Ministry of Education, Hainan University, Haikou 570228, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8830-4954","authenticated-orcid":false,"given":"Xiaopeng","family":"Zhu","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9358-5970","authenticated-orcid":false,"given":"Sulan","family":"Luo","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China"},{"name":"Key Laboratory of Tropical Biological Resources of Ministry of Education, Hainan University, Haikou 570228, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1038\/nrn731","article-title":"Ion channel structure: Emerging structure of the Nicotinic Acetylcholine receptors","volume":"3","author":"Karlin","year":"2002","journal-title":"Nat. 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