{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T18:58:21Z","timestamp":1780513101345,"version":"3.54.1"},"reference-count":43,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:00:00Z","timestamp":1760054400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"International Science and Technology Cooperation Key Special Project of the National Key Research and Development Program","award":["2023YFE0104500"],"award-info":[{"award-number":["2023YFE0104500"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["31702312"],"award-info":[{"award-number":["31702312"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background: The Tibetan Plateau, which is known for its high elevation and low oxygen levels, presents a challenging environment for its inhabitants. To adapt to these hypoxic conditions, species of Schizothoracine, a subfamily of Cyprinidae, have developed unique physiological mechanisms and functions. Transforming growth factor-\u03b2 (TGF-\u03b2) is a multifunctional cytokine involved in the regulation of cell growth, differentiation, apoptosis, and the cellular immune response. However, its specific role in adaptation to hypoxia remains poorly understood. Methods: In this study, we aimed to characterize the TGF-\u03b21 gene in Gymnocypris dobula (Gd) and Schizothorax prenanti (Sp) and to test whether TGF-\u03b21 contributes to hypoxia adaptation in plateau Schizothoracine fish. The predicted protein for Gd-TGF-\u03b21 contains several primary domains, including cwf21 (cdc5 protein 21), GYF (Glycine-Tyrosine-Phenylalanine), FN1 (Fibronectin 1), a conservative domain, and a signal peptide. Results: The results of tissue distribution revealed that the mRNA level of TGF-\u03b21 in brain, heart, muscle, skin, gills, and spleen\u2014which are key tissues involved in oxygen sensing, transport, and physiological adaptation to hypoxic environments\u2014was significantly lower in G. dobula than that in S. prenanti. Western blotting analysis revealed that the expression of activated TGF-\u03b21 in G. dobula was significantly higher than that in S. prenanti. To investigate whether TGF-\u03b21 in G. dobula possesses hypoxic adaptive features, Gd-TGF-\u03b21 and Sp-TGF-\u03b21 were cloned into an expression vector and transfected into 293-T cells, which are widely used due to their ease of culture, high transfectability, and well-characterized properties. We found that the survival rate of cells transfected with Gd-TGF-\u03b21 was significantly higher than that of cells transfected with Sp-TGF-\u03b21 after hypoxia treatment. Conclusions: These findings suggest that G. dobula may promote hypoxic adaptation through the activation and increased expression of TGF-\u03b21. Changes in TGF-\u03b21 expression may play a role in the adaptation of G. dobula to hypoxic conditions.<\/jats:p>","DOI":"10.3390\/genes16101176","type":"journal-article","created":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T12:32:47Z","timestamp":1760099567000},"page":"1176","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Analysis of the TGF-\u03b21 of a Tibetan Plateau Schizothoracine Fish (Gymnocypris dobula) Revealed Enhanced Cytoprotection in Hypoxic Environments"],"prefix":"10.3390","volume":"16","author":[{"given":"Ziyu","family":"Le","sequence":"first","affiliation":[{"name":"College of Marine Living Resource Sciences and Management, Shanghai Ocean University, Shanghai 201306, China"},{"name":"School of Life Sciences, Fudan University, Shanghai 200438, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohui","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Marine Living Resource Sciences and Management, Shanghai Ocean University, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Liu","sequence":"additional","affiliation":[{"name":"Marine Biomedical Science and Technology Innovation Platform of Lin-Gang Special Area, Ministry of Agriculture and Rural Affairs, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0351-1765","authenticated-orcid":false,"given":"Qianghua","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Marine Living Resource Sciences and Management, Shanghai Ocean University, Shanghai 201306, China"},{"name":"National Distant-Water Fisheries Engineering Research Center, Shanghai Ocean University, Shanghai 201306, China"},{"name":"Key Laboratory of Sustainable Exploitation of Oceanic Fisheries Resources, Ministry of Education, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Congcong","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Marine Living Resource Sciences and Management, Shanghai Ocean University, Shanghai 201306, China"},{"name":"Marine Biomedical Science and Technology Innovation Platform of Lin-Gang Special Area, Ministry of Agriculture and Rural Affairs, Shanghai 201306, China"},{"name":"National Distant-Water Fisheries Engineering Research Center, Shanghai Ocean University, Shanghai 201306, China"},{"name":"Key Laboratory of Sustainable Exploitation of Oceanic Fisheries Resources, Ministry of Education, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2295","DOI":"10.1139\/f75-268","article-title":"Minimal dissolved oxygen requirements of aquatic life with emphasis on Canadian species: A review","volume":"32","author":"Davis","year":"1975","journal-title":"J. Fish. Res. Board Can."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1139\/a06-006","article-title":"The effects of hypoxia on fishes: From ecological relevance to physiological effects","volume":"15","author":"Pollock","year":"2007","journal-title":"Environ. Rev."},{"key":"ref_3","first-page":"5073","article-title":"Water Environmental Characteristics and Water Quality Assessment of Lakes in Tibetan Plateau","volume":"43","author":"Liu","year":"2022","journal-title":"Environ. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1007\/s10695-019-00694-7","article-title":"Hypoxia-inducible factor 1\u03b1 from a high-altitude fish enhances cytoprotection and elevates nitric oxide production in hypoxic environment","volume":"46","author":"Wang","year":"2020","journal-title":"Fish Physiol. Biochem."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Xu, Q.H., Zhang, C., Zhang, D.S., Jiang, H.P., Peng, S.H., Liu, Y., Zhao, K., Wang, C.C., and Chen, L.B. (2016). Analysis of the erythropoietin of a Tibetan Plateau Schizothoracine fish (Gymnocypris dobula) reveals enhanced cytoprotection function in hypoxic environments. BMC Evol. Biol., 16.","DOI":"10.1186\/s12862-015-0581-0"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1007\/s10695-022-01080-6","article-title":"Characterization of EPO H131S as a key mutation site in the hypoxia-adaptive evolution of Gymnocypris dobula","volume":"48","author":"Wang","year":"2022","journal-title":"Fish Physiol. Biochem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/S0020-0255(02)00231-1","article-title":"Evolutionary analysis of functional divergence in TGF-\u03b2 signaling pathway","volume":"145","author":"Gu","year":"2002","journal-title":"Inf. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1006\/fsim.1999.0239","article-title":"Molecular isolation and characterisation of carp transforming growth factor beta 1 from activated leucocytes","volume":"10","author":"Zhan","year":"2000","journal-title":"Fish Shellfish Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1931","DOI":"10.1210\/en.2002-0126","article-title":"Cloning of Transforming growth factor-beta 1(TGF-beta1) and its type I receptor from zebrafish ovary and role of TGF-beta l in ocyte maturation","volume":"144","author":"Gurneet","year":"2003","journal-title":"Endocrinology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1792","DOI":"10.1016\/j.molimm.2007.09.027","article-title":"Grass carp transforming growth factor-beta 1 (TGF beta1): Molecular cloning, tissue distribution and immunobiological activity in teleost peripheral blood lymphocytes","volume":"45","author":"Yang","year":"2008","journal-title":"Mol. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"S53","DOI":"10.1046\/j.1523-1755.2000.07709.x","article-title":"Mechanism of transforming growth factor-beta1 signaling","volume":"58","author":"Choi","year":"2000","journal-title":"Kidney Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1007\/s12264-008-0130-6","article-title":"Neuropeptide y promotes TGF-\u03b21 production in raw264.7 cells by activating PI3K pathway via Y1 receptor","volume":"24","author":"Zhou","year":"2008","journal-title":"Neurosci. Bull."},{"key":"ref_13","unstructured":"Williams, E.J. (2001). Secretion and Activation of TGF-\u03b21 by Hepatic Stellate Cells and Its Contribution to Liver Fibrosis. [Ph.D. Thesis, University of Southampton]. Available online: https:\/\/www.researchgate.net\/publication\/361923012."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1385\/CBB:39:1:13","article-title":"TGF-b1 neuroprotective factor in cerebral ischemia","volume":"39","author":"Dhandapani","year":"2003","journal-title":"Cell Biochem. Biophys."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1081\/JDI-120006767","article-title":"Transforming growth factor-beta1 (TGF-beta1): A potential recovery signal in the post-ischemic kidney","volume":"24","author":"Docherty","year":"2002","journal-title":"Ren. Fail."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S1043-4666(03)00253-9","article-title":"Hypoxia modulates the effects of transforming growth factorbeta isoforms on matrix-formation by primary human lung fibroblasts","volume":"24","author":"Papakonstantinou","year":"2003","journal-title":"Cytokine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1647","DOI":"10.1097\/01.TP.0000063128.86981.B2","article-title":"Expression of transforming growth factor-beta1 and hypoxia-inducible factor-1 alpha in an experimental model of kidney transplantation","volume":"75","author":"Lario","year":"2003","journal-title":"Transplantation"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1006\/jsre.2001.6128","article-title":"Hypoxia regulates osteoblast gene expression","volume":"99","author":"Warren","year":"2001","journal-title":"Surg. Res."},{"key":"ref_19","first-page":"658","article-title":"Hypoxia potentiates transforming growth factor-beta expression of hepatocyte during the cirrhotic condition in rat liver","volume":"24","author":"Jeong","year":"2004","journal-title":"Liver Int. Off. J. Int. Assoc. Study Liver"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1016\/j.aquatox.2008.08.021","article-title":"Hypoxia alters gene expression in the gonads of zebrafish (Danio rerio)","volume":"95","author":"Martinovic","year":"2009","journal-title":"Aquat. Toxicol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.cbpc.2006.06.012","article-title":"Multiple tissue gene expression analyses in Japanese medaka (Oryzias latipes) exposed to hypoxia","volume":"145","author":"Ju","year":"2007","journal-title":"Comp. Biochem. Physiol. C Toxicol. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhang, H., Hu, Z., Li, R., Wang, Y., Zhou, J., Xu, H., Wang, G., Qiu, X., and Wang, X. (2022). Metabolomic Analysis of the Takifugu obscurus Gill under Acute Hypoxic Stress. Animals, 12.","DOI":"10.3390\/ani12192611"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/S0015-0282(02)03146-1","article-title":"Hypoxia-indued irreversible up-regulation of type 1 collagen and transforming growth factor-beta l in human peritoneal fibroblasts","volume":"78","author":"Sand","year":"2002","journal-title":"Fertil. Steril."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1002\/pros.10279","article-title":"Increased growth factor production in a human prostatic stromal cell culture model caused by hypoxia","volume":"57","author":"Berger","year":"2003","journal-title":"Prostate"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1016\/j.fsi.2012.11.011","article-title":"Transforming growth factor-\u03b21b: A second TGF-\u03b21 paralogue in the rainbow trout (Oncorhynchus mykiss) that has a lower constitutive expression but is more responsive to immune stimulation","volume":"34","author":"Maehr","year":"2013","journal-title":"Fish Shellfish Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1577\/H06-010.1","article-title":"Acute hypoxia\u2013reperfusion triggers immunocompromise in Nile tilapia","volume":"19","author":"Choi","year":"2007","journal-title":"J. Aquat. Anim. Health"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/S0145-305X(94)90357-3","article-title":"Effects of transforming growth factor \u03b21 on rainbow trout Oncorhynchus mykiss macrophage respiratory burst activity","volume":"18","author":"Jang","year":"1994","journal-title":"Dev. Comp. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Aksakal, E., and Ekinci, D. (2021). Effects of hypoxia and hyperoxia on growth parameters and transcription levels of growth, immune system and stress related genes in rainbow trout. Comp. Biochem. Physiol.\u2014Part A Mol. Integr. Physiol., 262.","DOI":"10.1016\/j.cbpa.2021.111060"},{"key":"ref_29","first-page":"6582","article-title":"HIF-1\u03b1 activates transforming growth factor-\u03b21\/Smad signaling in rat hepatic stellate cells under hypoxic conditions","volume":"17","author":"Wang","year":"2018","journal-title":"Mol. Med. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"24171","DOI":"10.1074\/jbc.M604507200","article-title":"Transforming growth factor \u03b21 induces hypoxia-inducible factor-1 stabilization through selective inhibition of PHD2 expression","volume":"281","author":"McMahon","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_31","unstructured":"(2018). Laboratory Animal\u2014Guideline for Ethical Review of Animal Welfare (Standard No. GB\/T 35892-2018)."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Chen, X., Yu, Y., Gao, T., Liu, Z., Chen, S., and Jia, Y. (2025). Determination of Stable Reference Genes for Gene Expression Analysis in Black Rockfish (Sebastes schlegelii) Under Hypoxia Stress. Genes, 16.","DOI":"10.3390\/genes16010009"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1007\/s40071-014-0061-y","article-title":"Identification of candidate reference genes for qRT-PCR under hypoxic stress in Clarias batrachus (Linnaeus, 1758)","volume":"6","author":"Mohindra","year":"2014","journal-title":"Int. Aquat. Res."},{"key":"ref_34","first-page":"2103","article-title":"Identification of suitable reference genes for qRT-PCR analysis in yellow catfish Pelteobagrus fulvidraco under hypoxia stress","volume":"49","author":"Hu","year":"2018","journal-title":"Aquac. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1287","DOI":"10.1007\/s10695-016-0218-2","article-title":"Morpho-histology of head kidney of female catfish Heteropneustes fossilis: Seasonal variations in melano-macrophage centers, melanin contents and effects of lipopolysaccharide and dexamethasone on melanins","volume":"42","author":"Kumar","year":"2016","journal-title":"Fish Physiol. Biochem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"D384","DOI":"10.1093\/nar\/gkac1096","article-title":"The conserved domain database in 2023","volume":"51","author":"Wang","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_37","first-page":"1797","article-title":"GYF Domain Proteomics Reveals Interaction Sites in Known and Novel Target Proteins","volume":"11","author":"Michael","year":"2005","journal-title":"Mol. Cell. Proteom."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1042\/CS20110086","article-title":"TGF-\u03b2\/TGF-\u03b2 receptor system and its role in physiological and pathological conditions","volume":"121","author":"Quintanilla","year":"2011","journal-title":"Clin. Sci."},{"key":"ref_39","unstructured":"Hou, S.F. (2014). Expression Characteristics and Functional Analysis of the Hypoxia Response Gene TGF-\u03b21 in Zebrafish. [Master\u2019s Thesis, Central China Normal University]. Available online: https:\/\/kns.cnki.net\/kcms2\/article\/abstract?v=F0lYaGTXfx-o0RcxQIfOC_Mb4f5RwCBQVSy_NB2rH-KDBIkQ515rhMqnJ7vQJf5T-UTHqce1meM1PQTHYGrK2i-Ij9ICnKDzPUIzhIy5EibdHYQtWIlGy8hWDtSTun-hSiHjCDYSYeUatzUR4_ZJL3gmHnQwLQYnerb1wXFw7zw3bBvo7OXjxm3pxfFnGuFg&uniplatform=NZKPT&language=CHS."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3966","DOI":"10.1016\/j.febslet.2009.10.036","article-title":"Correlation of mRNA and Protein in Complex Biological Samples","volume":"583","author":"Maier","year":"2009","journal-title":"FEBS Lett."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1038\/nrg3185","article-title":"Insights into the Regulation of Protein Abundance from Proteomic and Transcriptomic Analyses","volume":"13","author":"Vogel","year":"2012","journal-title":"Nat. Rev. Genet."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1038\/nature10152","article-title":"Latent TGF-\u03b2 structure and activation","volume":"474","author":"Shi","year":"2011","journal-title":"Nature"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"31557","DOI":"10.1074\/jbc.M503714200","article-title":"Involvement of transforming growth factor-\u03b21 signaling in hypoxia-induced tolerance to glucose starvation","volume":"280","author":"Suzuki","year":"2005","journal-title":"J. Biol. Chem."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/16\/10\/1176\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T04:22:55Z","timestamp":1760242975000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/16\/10\/1176"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,10]]},"references-count":43,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["genes16101176"],"URL":"https:\/\/doi.org\/10.3390\/genes16101176","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,10]]}}}