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Given the lack of disease-modifying cures, current therapies are limited to symptomatic relief. Here, we investigated the neurotrophic potential of the skin secretion (SS) from Rhinella schneideri, its polar fraction (PF) and nonpolar (NPF) fraction, and respective subfractions on the morphology of neuron-like cells. Following initial H2O-CH2CL2 partitioning, PF and NPF subfractions were isolated via RP-HPLC. Chemical characterization using LC-MS-IT-TOF identified eight distinct molecules, notably bufotenine and marinobufagin. Cytotoxicity screening established safe working concentrations (100\u2013250 ng\/mL for SS\/PF; 250\u2013500 ng\/mL for NPF and subfractions) for downstream morphological evaluations using High Content Screening (HCS). The subfraction polar 5 (SfP5) elicited a robust neurotrophic response, significantly enhancing all assessed morphometric parameters: total neurite outgrowth (+72%), branching points (+120%), maximum process length (+60%), and total number of processes (+35%). Our data show that Rhinella schneideri SS contains molecules that improve in vitro neuronal networks, serving as a promising source for preliminary screening of neuroprotective effects.<\/jats:p>","DOI":"10.3390\/toxins18060271","type":"journal-article","created":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T18:35:05Z","timestamp":1782239705000},"page":"271","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Enhancing Neuronal Networks with Rhinella schneideri Skin Secretion Molecules: Implications for Neurodegenerative Disorders"],"prefix":"10.3390","volume":"18","author":[{"given":"Giovanna Arruda","family":"Caires","sequence":"first","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias-Toxinologia do Instituto Butantan, Sao Paulo 05503-900, Brazil"},{"name":"Laboratory of Genetics, Butantan Institute, Sao Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabela Souza","family":"Pereira","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias-Toxinologia do Instituto Butantan, Sao Paulo 05503-900, Brazil"},{"name":"Laboratory of Genetics, Butantan Institute, Sao Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8933-4474","authenticated-orcid":false,"given":"Carlos","family":"DeOcesano-Pereira","sequence":"additional","affiliation":[{"name":"Center of Excellence in New Target Discovery, Butantan Institute, Sao Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2406-0860","authenticated-orcid":false,"given":"Daniel Carvalho","family":"Pimenta","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias-Toxinologia do Instituto Butantan, Sao Paulo 05503-900, Brazil"},{"name":"Laboratory of Biochemistry and Biophysics, Butantan Institute, Sao Paulo 05503-900, Brazil"},{"name":"Laboratory of Ecology and Evolution, Butantan Institute, Sao Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irina","family":"Kerkis","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias-Toxinologia do Instituto Butantan, Sao Paulo 05503-900, Brazil"},{"name":"Laboratory of Genetics, Butantan Institute, Sao Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7213-206X","authenticated-orcid":false,"given":"Juliana Mozer","family":"Sciani","sequence":"additional","affiliation":[{"name":"Laboratory of Natural Products, S\u00e3o Francisco University, Bragan\u00e7a Paulista 12916-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8197-2260","authenticated-orcid":false,"given":"Hugo","family":"Vigerelli","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias-Toxinologia do Instituto Butantan, Sao Paulo 05503-900, Brazil"},{"name":"Laboratory of Genetics, Butantan Institute, Sao Paulo 05503-900, Brazil"},{"name":"Center of Excellence in New Target Discovery, Butantan Institute, Sao Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1517\/14712598.2011.621940","article-title":"Venoms as a Platform for Human Drugs: Translating Toxins into Therapeutics","volume":"11","author":"King","year":"2011","journal-title":"Expert Opin. 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