{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T13:17:49Z","timestamp":1790774269197,"version":"4.1.0"},"reference-count":46,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2020,7,22]],"date-time":"2020-07-22T00:00:00Z","timestamp":1595376000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR19-CE20-0003-01"],"award-info":[{"award-number":["ANR19-CE20-0003-01"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"name":"E2S UPPA","award":["Junior chair 2018-2023"],"award-info":[{"award-number":["Junior chair 2018-2023"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Nowadays, aquaculture provides more than 50% of fish consumed worldwide but faces new issues that challenge its sustainability. One of them relies on the replacement of fish meal (FM) in aquaculture feeds by other protein sources without deeply affecting the whole organism\u2019s homeostasis. Multiple strategies have already been tested using in vivo approaches, but they hardly managed to cope with the multifactorial problems related to the complexities of fish biology together with new feed formulations. In this context, rainbow trout (RT) is particularly concerned by these problems, since, as a carnivorous fish, dietary proteins provide the amino acids required to supply most of its energetic metabolism. Surprisingly, we noticed that in vitro approaches considering RT cell lines as models to study RT amino acid metabolism were never previously used. Therefore, we decided to investigate if, and how, three major pathways described, in other species, to be regulated by amino acid and to control cellular homeostasis were functional in a RT cell line called RTH-149\u2014namely, the mechanistic Target Of Rapamycin (mTOR), autophagy and the general control nonderepressible 2 (GCN2) pathways. Our results not only demonstrated that these three pathways were functional in RTH-149 cells, but they also highlighted some RT specificities with respect to the time response, amino acid dependencies and the activation levels of their downstream targets. Altogether, this article demonstrated, for the first time, that RT cell lines could represent an interesting alternative of in vivo experimentations for the study of fish nutrition-related questions.<\/jats:p>","DOI":"10.3390\/cells9081754","type":"journal-article","created":{"date-parts":[[2020,7,23]],"date-time":"2020-07-23T11:26:01Z","timestamp":1595503561000},"page":"1754","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["RTH-149 Cell Line, a Useful Tool to Decipher Molecular Mechanisms Related to Fish Nutrition"],"prefix":"10.3390","volume":"9","author":[{"given":"Guillaume","family":"Morin","sequence":"first","affiliation":[{"name":"Universit\u00e9 de Pau et des Pays de l\u2019Adour, E2S UPPA, INRAE, NUMEA, 64310 Saint-P\u00e9e-sur-Nivelle, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karine","family":"Pinel","sequence":"additional","affiliation":[{"name":"Universit\u00e9 de Pau et des Pays de l\u2019Adour, E2S UPPA, INRAE, NUMEA, 64310 Saint-P\u00e9e-sur-Nivelle, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karine","family":"Dias","sequence":"additional","affiliation":[{"name":"Universit\u00e9 de Pau et des Pays de l\u2019Adour, E2S UPPA, INRAE, NUMEA, 64310 Saint-P\u00e9e-sur-Nivelle, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2202-1756","authenticated-orcid":false,"given":"Iban","family":"Seiliez","sequence":"additional","affiliation":[{"name":"Universit\u00e9 de Pau et des Pays de l\u2019Adour, E2S UPPA, INRAE, NUMEA, 64310 Saint-P\u00e9e-sur-Nivelle, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6017-7172","authenticated-orcid":false,"given":"Florian","family":"Beaumatin","sequence":"additional","affiliation":[{"name":"Universit\u00e9 de Pau et des Pays de l\u2019Adour, E2S UPPA, INRAE, NUMEA, 64310 Saint-P\u00e9e-sur-Nivelle, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,22]]},"reference":[{"key":"ref_1","unstructured":"FAO (2020). The State of World Fisheries and Aquaculture 2020: Sustainability in Action, The State of World Fisheries and Aquaculture (SOFIA); FAO."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15103","DOI":"10.1073\/pnas.0905235106","article-title":"Feeding aquaculture in an era of finite resources","volume":"106","author":"Naylor","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/S0044-8486(03)00422-8","article-title":"Almost total replacement of fish meal by plant protein sources in the diet of a marine teleost, the European seabass, Dicentrarchus labrax","volume":"230","author":"Kaushik","year":"2004","journal-title":"Aquaculture"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1079\/BJN20041176","article-title":"Dietary plant-protein substitution affects hepatic metabolism in rainbow trout (Oncorhynchus mykiss)","volume":"92","author":"Vilhelmsson","year":"2004","journal-title":"Br. J. Nutr."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1007\/s11306-018-1454-5","article-title":"Characterizing alternative feeds for rainbow trout (O-mykiss) by H-1 NMR metabolomics","volume":"14","author":"Roques","year":"2018","journal-title":"Metabolomics"},{"key":"ref_6","first-page":"164","article-title":"A review on replacing fish meal in aqua feeds using plant protein sources","volume":"6","author":"Daniel","year":"2018","journal-title":"Int. J. Fish. Aquat. Stud."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1016\/j.aquaculture.2004.01.006","article-title":"Effect of long-term feeding with a plant protein mixture based diet on growth and body\/fillet quality traits of large rainbow trout (Oncorhynchus mykiss)","volume":"236","author":"Parisi","year":"2004","journal-title":"Aquaculture"},{"key":"ref_8","first-page":"229","article-title":"The growth in vitro of single isolated tissue cells","volume":"9","author":"Sanford","year":"1948","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1084\/jem.97.5.695","article-title":"Studies on the propagation in vitro of poliomyelitis viruses","volume":"97","author":"Scherer","year":"1953","journal-title":"J. Exp. Med."},{"key":"ref_10","first-page":"932","article-title":"The in vitro cultivation of tissues of domestic and laboratory animals","volume":"18","author":"Madin","year":"1957","journal-title":"Am. J. Vet. Res."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"(1960). The Principles of Humane Experimental Technique. Med. J. Aust., 1, 500.","DOI":"10.5694\/j.1326-5377.1960.tb73127.x"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"25","DOI":"10.7171\/jbt.18-2902-002","article-title":"The Cellosaurus, a Cell-Line Knowledge Resource","volume":"29","author":"Bairoch","year":"2018","journal-title":"J. Biomol. Tech."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.1126\/science.135.3508.1065","article-title":"Established eurythermic line of fish cells in vitro","volume":"135","author":"Wolf","year":"1962","journal-title":"Science"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1007\/s11626-017-0142-5","article-title":"Invitromatics, invitrome, and invitroomics: Introduction of three new terms for in vitro biology and illustration of their use with the cell lines from rainbow trout","volume":"53","author":"Bols","year":"2017","journal-title":"In Vitro Cell. Dev. Biol. Anim."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1038\/s41556-018-0205-1","article-title":"mTOR as a central hub of nutrient signalling and cell growth","volume":"21","author":"Kim","year":"2019","journal-title":"Nat. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"14484","DOI":"10.1074\/jbc.273.23.14484","article-title":"Amino acid sufficiency and mTOR regulate p70 S6 kinase and eIF-4E BP1 through a common effector mechanism","volume":"273","author":"Hara","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1981","DOI":"10.1091\/mbc.e08-12-1248","article-title":"Nutrient-dependent mTORC1 association with the ULK1-Atg13-FIP200 complex required for autophagy","volume":"20","author":"Hosokawa","year":"2009","journal-title":"Mol. Biol. Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1016\/j.biopha.2018.05.007","article-title":"Autophagy in health and disease: A comprehensive review","volume":"104","author":"Saha","year":"2018","journal-title":"Biomed. Pharm."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.15252\/embr.201642195","article-title":"The integrated stress response","volume":"17","author":"Koryga","year":"2016","journal-title":"EMBO Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1007\/BF02618871","article-title":"Fish cell lines: Establishment and characterization of nine cell lines from salmonids","volume":"20","author":"Lannan","year":"1984","journal-title":"In Vitro"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"10774","DOI":"10.1073\/pnas.1504276112","article-title":"The amino acid sensor GCN2 inhibits inflammatory responses to apoptotic cells promoting tolerance and suppressing systemic autoimmunity","volume":"112","author":"Ravishankar","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/15548627.2015.1100356","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)","volume":"12","author":"Klionsky","year":"2016","journal-title":"Autophagy"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.aquaculture.2010.03.012","article-title":"Skeletal muscle growth dynamics and expression of related genes in white and red muscles of rainbow trout fed diets with graded levels of a mixture of plant protein sources as substitutes for fishmeal","volume":"303","author":"Medale","year":"2010","journal-title":"Aquaculture"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1017\/S0007114514001226","article-title":"Dietary methionine availability affects the main factors involved in muscle protein turnover in rainbow trout (Oncorhynchus mykiss)","volume":"112","author":"Belghit","year":"2014","journal-title":"Br. J. Nutr."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1007\/s00726-010-0533-3","article-title":"Integration of insulin and amino acid signals that regulate hepatic metabolism-related gene expression in rainbow trout: Role of TOR","volume":"39","author":"Lansard","year":"2010","journal-title":"Amino Acids"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1016\/j.cbpb.2010.06.011","article-title":"An in vivo and in vitro assessment of autophagy-related gene expression in muscle of rainbow trout (Oncorhynchus mykiss)","volume":"157","author":"Seiliez","year":"2010","journal-title":"Comp. Biochem. Physiol. Part. B: Biochem. Mol. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1080\/15548627.2015.1117732","article-title":"Looking at the metabolic consequences of the colchicine-based in vivo autophagic flux assay","volume":"12","author":"Seiliez","year":"2016","journal-title":"Autophagy"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1007\/BF02631451","article-title":"Cultured Trout Liver-Cells-Utilization of Substrates and Response to Hormones","volume":"30A","author":"Segner","year":"1994","journal-title":"In Vitro Cell. Dev. Biol.-Anim"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Froehlich, J.M., Seiliez, I., Gabillard, J.-C., and Biga, P.R. (2014). Preparation of Primary Myogenic Precursor Cell\/Myoblast Cultures from Basal Vertebrate Lineages. J. Vis. Exp., e51354.","DOI":"10.3791\/51354-v"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1002\/jez.1402710603","article-title":"In-Vitro Hepatocyte Metabolism of Alanine and Glucose and the Response to Insulin in Fed and Fasted Rainbow-Trout","volume":"271","author":"Pereira","year":"1995","journal-title":"J. Exp. Zool."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/0005-2760(95)00211-1","article-title":"The desaturation and elongation of linolenic acid and eicosapentaenoic acid by hepatocytes and liver microsomes from rainbow trout (Oncorhynchus mykiss) fed diets containing fish oil or olive oil","volume":"1299","author":"Buzzi","year":"1996","journal-title":"Biochim. Biophys. Acta-Lipids Lipid Metab."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"R1615","DOI":"10.1152\/ajpregu.00114.2010","article-title":"Paralogs of Atlantic salmon myoblast determination factor genes are distinctly regulated in proliferating and differentiating myogenic cells","volume":"298","author":"Bower","year":"2010","journal-title":"Am. J. Physiol.-Regul. Integr. Comp. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.cbpa.2013.10.020","article-title":"Characterisation and expression of myogenesis regulatory factors during in vitro myoblast development and in vivo fasting in the gilthead sea bream (Sparus aurata)","volume":"167","author":"Codina","year":"2014","journal-title":"Comp. Biochem. Physiol. A-Mol. Integr. Physiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.cbpc.2014.05.003","article-title":"In vitro and in vivo effects of phytoestrogens on protein turnover in rainbow trout (Oncorhynchus mykiss) white muscle","volume":"165","author":"Cleveland","year":"2014","journal-title":"Comp. Biochem. Physiol. C-Toxicol. Pharm."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"R341","DOI":"10.1152\/ajpregu.00516.2009","article-title":"Effects of insulin-like growth factor-I, insulin, and leucine on protein turnover and ubiquitin ligase expression in rainbow trout primary myocytes","volume":"298","author":"Cleveland","year":"2010","journal-title":"Am. J. Physiol.-Regul. Integr. Comp. Physiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"35106","DOI":"10.1074\/jbc.M806874200","article-title":"C\/EBP Homology Protein (CHOP) Interacts with Activating Transcription Factor 4 (ATF4) and Negatively Regulates the Stress-dependent Induction of the Asparagine Synthetase Gene","volume":"283","author":"Su","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"7683","DOI":"10.1093\/nar\/gkt563","article-title":"The eIF2 alpha\/ATF4 pathway is essential for stress-induced autophagy gene expression","volume":"41","author":"Maurin","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1016\/j.molcel.2016.09.037","article-title":"An Autophagic Flux Probe that Releases an Internal Control","volume":"64","author":"Kaizuka","year":"2016","journal-title":"Mol. Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.cell.2008.11.044","article-title":"Bidirectional transport of amino acids regulates mTOR and autophagy","volume":"136","author":"Nicklin","year":"2009","journal-title":"Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.molcel.2019.07.021","article-title":"mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux","volume":"76","author":"Beaumatin","year":"2019","journal-title":"Mol. Cell"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"364","DOI":"10.4161\/auto.18863","article-title":"Amino acids downregulate the expression of several autophagy-related genes in rainbow trout myoblasts","volume":"8","author":"Seiliez","year":"2012","journal-title":"Autophagy"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"R329","DOI":"10.1152\/ajpregu.00146.2008","article-title":"An in vivo and in vitro assessment of TOR signaling cascade in rainbow trout (Oncorhynchus mykiss)","volume":"295","author":"Seiliez","year":"2008","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"75","DOI":"10.3945\/jn.110.124511","article-title":"L-leucine, L-methionine, and L-lysine are involved in the regulation of intermediary metabolism-related gene expression in rainbow trout hepatocytes","volume":"141","author":"Lansard","year":"2011","journal-title":"J. Nutr."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e19960","DOI":"10.7554\/eLife.19960","article-title":"Dynamics of mTORC1 activation in response to amino acids","volume":"5","author":"Manifava","year":"2016","journal-title":"Elife"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1685","DOI":"10.1007\/s00726-018-2640-5","article-title":"Protein content and amino acid composition of commercially available plant-based protein isolates","volume":"50","author":"Gorissen","year":"2018","journal-title":"Amino Acids"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"101","DOI":"10.2527\/jas1983.561101x","article-title":"Requirement for dietary lysine and arginine by fry of rainbow trout","volume":"56","author":"Ketola","year":"1983","journal-title":"J. Anim. Sci."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/8\/1754\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:50:44Z","timestamp":1760176244000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/8\/1754"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,22]]},"references-count":46,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,8]]}},"alternative-id":["cells9081754"],"URL":"https:\/\/doi.org\/10.3390\/cells9081754","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,22]]}}}