{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T03:52:12Z","timestamp":1791604332728,"version":"4.3.4"},"reference-count":42,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2021YFA0805000"],"award-info":[{"award-number":["2021YFA0805000"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["82241201"],"award-info":[{"award-number":["82241201"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Jiangsu Double Innovation Talent Program","award":["JSSCRC2021567"],"award-info":[{"award-number":["JSSCRC2021567"]}]},{"name":"Jiangsu Cardiovascular Medicine Innovation Center","award":["CXZX202210"],"award-info":[{"award-number":["CXZX202210"]}]},{"DOI":"10.13039\/501100012246","name":"Priority Academic Program Development of Jiangsu Higher Education Institutions","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100012246","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Congenital valve defects account for a substantial proportion of cardiovascular malformations, yet the molecular mechanisms orchestrating cardiac valve development remain incompletely elucidated. While Bone morphogenetic protein (BMP) signaling is essential for valvulogenesis, the specific contributions of individual BMP ligands, particularly the teleost-specific bmp16, have not been characterized. Using the CRISPR\/Cas9 system, we generated a bmp16 null knockout and delineated critical roles of this ligand in valvular morphogenesis. bmp16 knockout embryos display a significant reduction in Sox9-positive valvular cells and exhibit severely dysplastic arterial valves, characterized by increased interleaflet distance, thickened leaflets, and shortened leaflet lengths. These morphological abnormalities correlate with impaired valve function, culminating in progressive blood regurgitation, ventricular dilation, and pericardial edema. Mechanistically, loss of bmp16 or pharmacological inhibition of BMP signaling significantly downregulates notch1b expression in developing valves, while pharmacological activation of Notch signaling rescues the regurgitation phenotype in bmp16 mutants. Collectively, our findings establish bmp16 as a novel regulator of valve development and uncover a functional BMP-Notch signaling axis required for vertebrate valvulogenesis, providing new insights into the molecular mechanisms that govern cardiac valve formation and pathogenesis.<\/jats:p>","DOI":"10.3390\/ijms27052111","type":"journal-article","created":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T16:58:18Z","timestamp":1771952298000},"page":"2111","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Bmp16 Regulates Arterial Valve Morphogenesis Through Modulation of Notch Signaling in Zebrafish"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1506-2469","authenticated-orcid":false,"given":"Mengting","family":"Xu","sequence":"first","affiliation":[{"name":"Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University, Suzhou Medical College, Soochow University, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunmiao","family":"Ma","sequence":"additional","affiliation":[{"name":"Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University, Suzhou Medical College, Soochow University, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengxin","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University, Suzhou Medical College, Soochow University, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yueqiu","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University, Suzhou Medical College, Soochow University, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zongyi","family":"Duan","sequence":"additional","affiliation":[{"name":"Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University, Suzhou Medical College, Soochow University, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenya","family":"Shen","sequence":"additional","affiliation":[{"name":"Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University, Suzhou Medical College, Soochow University, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1838-0836","authenticated-orcid":false,"given":"Yanchao","family":"Han","sequence":"additional","affiliation":[{"name":"Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University, Suzhou Medical College, Soochow University, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1038\/s41569-022-00749-y","article-title":"Changing epidemiology of congenital heart disease: Effect on outcomes and quality of care in adults","volume":"20","author":"Liu","year":"2023","journal-title":"Nat. Rev. Cardiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2899","DOI":"10.1007\/s00018-012-1197-9","article-title":"Co-ordinating Notch, BMP, and TGF-beta signaling during heart valve development","volume":"70","author":"Garside","year":"2013","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.1016\/S0140-6736(23)02755-1","article-title":"Valvular heart disease: From mechanisms to management","volume":"403","author":"Praz","year":"2024","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1080\/00015385.2020.1746053","article-title":"Aortic valve: Anatomy and structure and the role of vasculature in the degenerative process","volume":"76","author":"Katsi","year":"2021","journal-title":"Acta Cardiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"dev183020","DOI":"10.1242\/dev.183020","article-title":"Mechanisms of heart valve development and disease","volume":"147","author":"Yutzey","year":"2020","journal-title":"Development"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1431","DOI":"10.1161\/01.RES.0000224114.65109.4e","article-title":"Extracellular matrix remodeling and organization in developing and diseased aortic valves","volume":"98","author":"Hinton","year":"2006","journal-title":"Circ. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1093\/cvr\/cvae230","article-title":"Zebrafish arterial valve development occurs through direct differentiation of second heart field progenitors","volume":"121","author":"Derrick","year":"2025","journal-title":"Cardiovasc. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1633","DOI":"10.1242\/dev.030924","article-title":"Distinct phases of cardiomyocyte differentiation regulate growth of the zebrafish heart","volume":"136","author":"Clijsters","year":"2009","journal-title":"Development"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Wang, Y., Fang, Y., Lu, P., Wu, B., and Zhou, B. (2021). NOTCH signaling in aortic valve development and calcific aortic valve disease. Front. Cardiovasc. Med., 8.","DOI":"10.3389\/fcvm.2021.682298"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1038\/nature03940","article-title":"Mutations in NOTCH1 cause aortic valve disease","volume":"437","author":"Garg","year":"2005","journal-title":"Nature"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.ceb.2018.05.007","article-title":"The mechanobiology of zebrafish cardiac valve leaflet formation","volume":"55","author":"Paolini","year":"2018","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"107883","DOI":"10.1016\/j.celrep.2020.107883","article-title":"Antagonistic activities of Vegfr3\/Flt4 and Notch1b Fine-tune mechanosensitive signaling during Zebrafish cardiac Valvulogenesis","volume":"32","author":"Fontana","year":"2020","journal-title":"Cell. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5601","DOI":"10.1242\/dev.02156","article-title":"Bmp2 is essential for cardiac cushion epithelial-mesenchymal transition and myocardial patterning","volume":"132","author":"Ma","year":"2005","journal-title":"Development"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2362","DOI":"10.1101\/gad.1124803","article-title":"An essential role of Bmp4 in the atrioventricular septation of the mouse heart","volume":"17","author":"Jiao","year":"2003","journal-title":"Genes Dev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3200","DOI":"10.1002\/dvdy.21743","article-title":"BMP4 is required in the anterior heart field and its derivatives for endocardial cushion remodeling, outflow tract septation, and semilunar valve development","volume":"237","author":"McCulley","year":"2008","journal-title":"Dev. Dyn."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1002\/dvg.20511","article-title":"Bmp2 and Bmp4 genetically interact to support multiple aspects of mouse development including functional heart development","volume":"47","author":"Uchimura","year":"2009","journal-title":"Genesis"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1006\/dbio.2001.0284","article-title":"Bmp6 and Bmp7 are required for cushion formation and septation in the developing mouse heart","volume":"235","author":"Kim","year":"2001","journal-title":"Dev. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1007\/s00018-015-2024-x","article-title":"Comparative analysis of zebrafish bone morphogenetic proteins 2, 4 and 16: Molecular and evolutionary perspectives","volume":"73","author":"Marques","year":"2016","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Feiner, N., Begemann, G., Renz, A.J., Meyer, A., and Kuraku, S. (2009). The origin of bmp16, a novel Bmp2\/4 relative, retained in teleost fish genomes. BMC Evol. Biol., 9.","DOI":"10.1186\/1471-2148-9-277"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3136","DOI":"10.1038\/s41598-019-40055-1","article-title":"Asymmetric paralog evolution between the \u201ccryptic\u201d gene Bmp16 and its well-studied sister genes Bmp2 and Bmp4","volume":"9","author":"Feiner","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1002\/dvdy.22567","article-title":"Dynamic smad-mediated BMP signaling revealed through transgenic zebrafish","volume":"240","author":"Collery","year":"2011","journal-title":"Dev. Dyn."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4883","DOI":"10.1074\/jbc.M111023200","article-title":"Identification and functional characterization of distinct critically important bone morphogenetic protein-specific response elements in the Id1 promoter","volume":"277","author":"Korchynskyi","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1002\/dvg.20402","article-title":"Real time monitoring of BMP Smads transcriptional activity during mouse development","volume":"46","author":"Monteiro","year":"2008","journal-title":"Genesis"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4653","DOI":"10.1242\/jcs.01337","article-title":"Spatio-temporal activation of Smad1 and Smad5 in vivo: Monitoring transcriptional activity of Smad proteins","volume":"117","author":"Monteiro","year":"2004","journal-title":"J. Cell Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"RP101151","DOI":"10.7554\/eLife.101151","article-title":"Contraction-induced endocardial id2b plays a dual role in regulating myocardial contractility and valve formation","volume":"13","author":"Chen","year":"2025","journal-title":"Elife"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s10456-019-09663-3","article-title":"A ribosomal DNA-hosted microRNA regulates zebrafish embryonic angiogenesis","volume":"22","author":"Shi","year":"2019","journal-title":"Angiogenesis"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2111","DOI":"10.1242\/dev.063933","article-title":"Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo","volume":"138","author":"Krueger","year":"2011","journal-title":"Development"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1363","DOI":"10.1038\/nm.1888","article-title":"BMP type I receptor inhibition reduces heterotopic [corrected] ossification","volume":"14","author":"Yu","year":"2008","journal-title":"Nat. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1002\/ar.23913","article-title":"Dynamic expression Profiles of Sox9 in embryonic, post natal, and adult Heart valve cell populations","volume":"302","author":"Gallina","year":"2019","journal-title":"Anat. Rec."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.ydbio.2012.03.023","article-title":"In vivo Wnt signaling tracing through a transgenic biosensor fish reveals novel activity domains","volume":"366","author":"Moro","year":"2012","journal-title":"Dev. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"151786","DOI":"10.1016\/j.bbrc.2025.151786","article-title":"Chemogenetic ablation and regeneration of arterial valve in zebrafish","volume":"762","author":"Duan","year":"2025","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2776","DOI":"10.1242\/dev.091876","article-title":"The zebrafish common cardinal veins develop by a novel mechanism: Lumen ensheathment","volume":"140","author":"Helker","year":"2013","journal-title":"Development"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1273","DOI":"10.1002\/stem.2843","article-title":"A novel 2-Phenylamino-Quinazoline-Based compound Expands the neural stem cell pool and Promotes the hippocampal neurogenesis and the Cognitive ability of adult mice","volume":"36","author":"Lu","year":"2018","journal-title":"Stem Cells"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1161\/CIRCRESAHA.109.201566","article-title":"Heart valve development: Regulatory networks in development and disease","volume":"105","author":"Combs","year":"2009","journal-title":"Circ. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1146\/annurev-genet-110711-155646","article-title":"Uncovering the molecular and cellular mechanisms of heart development using the zebrafish","volume":"46","author":"Staudt","year":"2012","journal-title":"Annu. Rev. Genet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"580","DOI":"10.1016\/j.ydbio.2006.03.043","article-title":"Bmp2 instructs cardiac progenitors to form the heart-valve-inducing field","volume":"295","author":"Tabin","year":"2006","journal-title":"Dev. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1670","DOI":"10.1126\/science.293.5535.1670","article-title":"UDP-glucose dehydrogenase required for cardiac valve formation in zebrafish","volume":"293","author":"Walsh","year":"2001","journal-title":"Science"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1161\/ATVBAHA.113.302071","article-title":"Conserved transcriptional regulatory mechanisms in aortic valve development and disease","volume":"34","author":"Wirrig","year":"2014","journal-title":"Arter. Thromb. Vasc. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3891","DOI":"10.1242\/dev.029264","article-title":"Tailbud-derived Bmp4 drives proliferation and inhibits maturation of zebrafish chordamesoderm","volume":"135","author":"Esterberg","year":"2008","journal-title":"Development"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1002\/jcp.21496","article-title":"Sox9, a key transcription factor of bone morphogenetic protein-2-induced chondrogenesis, is activated through BMP pathway and a CCAAT box in the proximal promoter","volume":"217","author":"Pan","year":"2008","journal-title":"J. Cell. Physiol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Wang, Y., Wu, B., Chamberlain, A.A., Lui, W., Koirala, P., Susztak, K., Klein, D., Taylor, V., and Zhou, B. (2013). Endocardial to myocardial notch-wnt-bmp axis regulates early heart valve development. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0060244"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1016\/j.devcel.2019.01.001","article-title":"Vitamin D stimulates cardiomyocyte proliferation and controls organ size and regeneration in zebrafish","volume":"48","author":"Han","year":"2019","journal-title":"Dev. 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