{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T13:08:38Z","timestamp":1781788118643,"version":"3.54.5"},"reference-count":49,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T00:00:00Z","timestamp":1766707200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science and Technology Council, Taiwan","award":["NSTC-114-2313-B-030-001"],"award-info":[{"award-number":["NSTC-114-2313-B-030-001"]}]},{"DOI":"10.13039\/501100013784","name":"Fu Jen Catholic University, Taiwan","doi-asserted-by":"publisher","award":["9991F02-1120232 and A0113210"],"award-info":[{"award-number":["9991F02-1120232 and A0113210"]}],"id":[{"id":"10.13039\/501100013784","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Collaborative Research Project between Fu Jen Catholic University Hospital and Fu Jen Catholic University, Taiwan","award":["CPL-202508003"],"award-info":[{"award-number":["CPL-202508003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Zebrafish is emerging as a model animal for phenotype-based drug screening. Drugs screened from the zebrafish platform have advanced into clinical trials, underscoring their translational potential. Amyotrophic lateral sclerosis is a progressive motor neurons (MN) degenerative disease with few approved drugs. Previously, supplementation with exogenous recombinant phosphoglycerate kinase 1 (Pgk1) was found to improve MN growth through its interaction with receptor Eno2. To bypass the high complexity and cost of full-length Pgk1 production, a short segment within Pgk1 (M08) was predicted as the key motif interacting with Eno2, and a zebrafish phenotypic screening platform was established to find the most neurotrophic compound(s) among M08 and its mutants. We first found that M08-injected zebrafish embryos significantly increased branched caudal primary MNs (CaPMNs). However, compared to M08 (59.20 \u00b1 1.80%), M039, among 17 mutants further screened, showed even more improvement of branched CaPMNs, up to 74.54 \u00b1 3.73%. Next, when we administered the M039 peptide to C9ORF72-knockdown ALS-like zebrafish embryos, it improved axonal growth and swimming ability. Then, we employed a cellular model as a secondary screen, and M039 exhibited improved neurite outgrowth of MN (NOMN) and reduced p-Cofilin in NSC34 neural cells grown in ALS-like condition. Therefore, by using a zebrafish MN phenotype as a primary screening platform, we identified a mutated short peptide M039 having the most pronounced positive effect on improving neurite growth among all 17 mutants in comparison to parental M08, demonstrating the feasibility of zebrafish screening as a cost-effective strategy for finding promising neuroprotective short peptides that serve as neurotherapeutic potentials.<\/jats:p>","DOI":"10.3390\/ijms27010281","type":"journal-article","created":{"date-parts":[[2025,12,29]],"date-time":"2025-12-29T05:28:19Z","timestamp":1766986099000},"page":"281","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Multilevel Screening Platform Utilizing Cellular and Zebrafish Models to Identify Short Peptides with High Improvement of Motor Neuron Growth"],"prefix":"10.3390","volume":"27","author":[{"given":"Bing-Chang","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Life Science, Fu Jen Catholic University, New Taipei City 242, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chun-Cheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Molecular and Cellular Biology, National Taiwan University, Taipei 106, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shan-Pin","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Molecular and Cellular Biology, National Taiwan University, Taipei 106, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8242-4939","authenticated-orcid":false,"given":"Huai-Jen","family":"Tsai","sequence":"additional","affiliation":[{"name":"Department of Life Science, Fu Jen Catholic University, New Taipei City 242, Taiwan"},{"name":"Institute of Molecular and Cellular Biology, National Taiwan University, Taipei 106, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1093\/af\/vfz020","article-title":"The use of zebrafish (Danio rerio) as biomedical models","volume":"9","author":"Teame","year":"2019","journal-title":"Anim. 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