{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T08:29:22Z","timestamp":1778056162792,"version":"3.51.4"},"reference-count":41,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2023,6,28]],"date-time":"2023-06-28T00:00:00Z","timestamp":1687910400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National High Level Hospital Clinical Research Funding","award":["2022-PUMCH-C-029"],"award-info":[{"award-number":["2022-PUMCH-C-029"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background: In vertebrates, the development of the inner ear is a delicate process, whereas its relating molecular pathways are still poorly understood. LMO4, an LIM domain-only transcriptional regulator, is drawing an increasing amount of interest for its multiple roles regarding human embryonic development and the modulation of ototoxic side effects of cisplatin including cochlear apoptosis and hearing loss. The aim of the present study is to further explore the role of lmo4a in zebrafish inner ear development and thus explore its functional role. Methods: The Spatial Transcript Omics DataBase was referred to in order to evaluate the expression of lmo4a during the first 24 h of zebrafish development. In situ hybridization was applied to validate and extend the expression profile of lmo4a to 3 days post-fertilization. The morpholino (MO) knockdown and CRISPR\/Cas9 knockout (KO) of lmo4a was applied. Morphological analyses of otic vesical, hair cells, statoacoustic ganglion and semicircular canals were conducted. The swimming pattern of lmo4a KO and MO zebrafish was tracked. In situ hybridization was further applied to verify the expression of genes of the related pathways. Rescue of the phenotype was attempted by blockage of the bmp pathway via heat shock and injection of Dorsomorphin. Results: lmo4a is constitutively expressed in the otic placode and otic vesicle during the early stages of zebrafish development. Knockdown and knockout of lmo4a both induced smaller otocysts, less hair cells, immature statoacoustic ganglion and malformed semicircular canals. Abnormal swimming patterns could be observed in both lmo4a MO and KO zebrafish. eya1 in preplacodal ectoderm patterning was downregulated. bmp2 and bmp4 expressions were found to be upregulated and extended in lmo4a morphants, and blockage of the Bmp pathway partially rescued the vestibular defects. Conclusions: We concluded that lmo4a holds a regulative effect on the Bmp pathway and is required for the normal development of zebrafish inner ear. Our study pointed out the conservatism of LMO4 in inner ear development between mammals and zebrafish as well as shed more light on the molecular mechanisms behind it. Further research is needed to distinguish the relationships between lmo4 and the Bmp pathway, which may lead to diagnostic and therapeutic approaches towards human inner ear malformation.<\/jats:p>","DOI":"10.3390\/genes14071371","type":"journal-article","created":{"date-parts":[[2023,6,29]],"date-time":"2023-06-29T00:49:47Z","timestamp":1687999787000},"page":"1371","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["lmo4a Contributes to Zebrafish Inner Ear and Vestibular Development via Regulation of the Bmp Pathway"],"prefix":"10.3390","volume":"14","author":[{"given":"Le","family":"Sun","sequence":"first","affiliation":[{"name":"Department of Otolaryngology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, #1 Shuaifuyuan, Dongcheng District, Beijing 100730, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lu","family":"Ping","sequence":"additional","affiliation":[{"name":"Chinese Academy of Medical Sciences and Peking Union Medical College, #9 Dongdan Santiao, Dongcheng District, Beijing 100050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruzhen","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bo","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaowei","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, #1 Shuaifuyuan, Dongcheng District, Beijing 100730, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1111\/joa.12308","article-title":"Form and function of the mammalian inner ear","volume":"228","author":"Ekdale","year":"2016","journal-title":"J. Anat."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1503","DOI":"10.1387\/ijdb.072422ba","article-title":"Patterning and cell fate in ear development","volume":"53","author":"Alsina","year":"2009","journal-title":"Int. J. Dev. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1631\/jzus.B1800084","article-title":"Bone morphogenetic proteins and inner ear development","volume":"20","author":"Ma","year":"2019","journal-title":"J. Zhejiang Univ. B"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/S0925-4773(01)00479-8","article-title":"Expression of BMP signalling pathway members in the developing zebrafish inner ear and lateral line","volume":"108","author":"Mowbray","year":"2001","journal-title":"Mech. Dev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3327","DOI":"10.1523\/JNEUROSCI.18-09-03327.1998","article-title":"Development of the mouse inner ear and origin of its sensory organs","volume":"18","author":"Morsli","year":"1998","journal-title":"J. Neurosci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"6454","DOI":"10.1523\/JNEUROSCI.16-20-06454.1996","article-title":"Sensory organ generation in the chick inner ear","volume":"16","author":"Wu","year":"1996","journal-title":"J. Neurosci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1002\/dvg.1020170109","article-title":"Ventral mesodermal patterning inXenopus embryos: Expression patterns and activities of BMP-2 and BMP-4","volume":"17","author":"Thomsen","year":"1995","journal-title":"Dev. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"11257","DOI":"10.1073\/pnas.95.19.11257","article-title":"Identification and characterization of LMO 4, an LMO gene with a novel pattern of expression during embryogenesis","volume":"95","author":"Kenny","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7659","DOI":"10.1073\/pnas.0502990102","article-title":"Overexpression of LMO4 induces mammary hyperplasia, promotes cell invasion, and is a predictor of poor outcome in breast","volume":"102","author":"Sum","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1016\/j.modgep.2006.02.005","article-title":"Differential expression of LIM domain-only (LMO) genes in the developing mouse inner ear","volume":"6","author":"Deng","year":"2006","journal-title":"Gene Expr. Patterns"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"10072","DOI":"10.1523\/JNEUROSCI.0352-14.2014","article-title":"LMO4 Functions as a Negative Regulator of Sensory Organ Formation in the Mammalian Cochlea","volume":"34","author":"Deng","year":"2014","journal-title":"J. Neurosci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.ydbio.2009.11.003","article-title":"Requirement for Lmo4 in the vestibular morphogenesis of mouse inner ear","volume":"338","author":"Deng","year":"2010","journal-title":"Dev. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3545","DOI":"10.1002\/jcb.26529","article-title":"CRISPR\/Cas9-mediated knockout of Lim-domain only four retards organ of Corti cell growth","volume":"119","author":"Rathinam","year":"2018","journal-title":"J. Cell. Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/j.redox.2016.10.016","article-title":"Targeting nitrative stress for attenuating cisplatin-induced downregulation of cochlear LIM domain only 4 and ototoxicity","volume":"10","author":"Jamesdaniel","year":"2016","journal-title":"Redox Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"15052","DOI":"10.1038\/cddiscovery.2015.52","article-title":"Cisplatin-induced apoptosis in auditory, renal, and neuronal cells is associated with nitration and downregulation of LMO4","volume":"1","author":"Rathinam","year":"2015","journal-title":"Cell Death Discov."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Jamesdaniel, S. (2014). Downstream Targets of Lmo4 Are Modulated by Cisplatin in the Inner Ear of Wistar Rats. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0115263"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"18674","DOI":"10.1074\/jbc.M111.297960","article-title":"Cisplatin-induced Ototoxicity Is Mediated by Nitroxidative Modification of Cochlear Proteins Characterized by Nitration of Lmo4","volume":"287","author":"Jamesdaniel","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2019","DOI":"10.1007\/s12035-020-02226-4","article-title":"Lmo4 Deficiency Enhances Susceptibility to Cisplatin-Induced Cochlear Apoptosis and Hearing Loss","volume":"58","author":"Rosati","year":"2021","journal-title":"Mol. Neurobiol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Chang, W., Lin, Z., Kulessa, H., Hebert, J., Hogan, B.L.M., and Wu, D.K. (2008). Bmp4 Is Essential for the Formation of the Vestibular Apparatus that Detects Angular Head Movements. PLoS Genet., 4.","DOI":"10.1371\/journal.pgen.1000050"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"dev174748","DOI":"10.1242\/dev.174748","article-title":"Genetic interactions support an inhibitory relationship between bone morphogenetic protein 2 and netrin 1 during semicircular canal formation","volume":"146","author":"Hwang","year":"2019","journal-title":"Development"},{"key":"ref_21","unstructured":"Mm, W. (1994). The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish Danio (Brachydanio) Rerio1994, Zebrafish International Resource Center."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2333","DOI":"10.1242\/dev.01806","article-title":"Transgenic zebrafish reveal stage-specific roles for Bmp signaling in ventral and posterior mesoderm development","volume":"132","author":"Pyati","year":"2005","journal-title":"Development"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.1016\/j.devcel.2022.04.009","article-title":"Spatiotemporal mapping of gene expression landscapes and developmental trajectories during zebrafish embryogenesis","volume":"57","author":"Liu","year":"2022","journal-title":"Dev. Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1007\/s004270050270","article-title":"The zebrafish eya1 gene and its expression pattern during embryogenesis","volume":"209","author":"Sahly","year":"1999","journal-title":"Dev. Genes Evol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/S0925-4773(96)00625-9","article-title":"Conservation of BMP signaling in zebrafish mesoderm patterning","volume":"61","author":"Nikaido","year":"1997","journal-title":"Mech. Dev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1203","DOI":"10.1242\/dev.119.4.1203","article-title":"Structure of the zebrafish snail1 gene and its expression in wild-type, spadetail and no tail mutant embryos","volume":"119","author":"Thisse","year":"1993","journal-title":"Development"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.devcel.2018.06.003","article-title":"A rapid method for directed gene knockout for screening in G0 zebrafish","volume":"46","author":"Wu","year":"2018","journal-title":"Dev. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1111\/j.1440-169X.2007.00964.x","article-title":"Role of Bone morphogenetic protein 4 in zebrafish semicircular canal development","volume":"49","author":"Omata","year":"2007","journal-title":"Dev. Growth Differ."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"580","DOI":"10.1006\/geno.1997.4978","article-title":"Relationship between the genomic organization and the overlapping embryonic expression patterns of the ZebrafishdlxGenes","volume":"45","author":"Ellies","year":"1997","journal-title":"Genomics"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3475","DOI":"10.1523\/JNEUROSCI.14-06-03475.1994","article-title":"Combinatorial expression of three zebrafish genes related to distal-less: Part of a homeobox gene code for the head","volume":"14","author":"Akimenko","year":"1994","journal-title":"J. Neurosci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/S0925-4773(00)00451-2","article-title":"Expression of three zebrafish Six4 genes in the cranial sensory placodes and the developing somites","volume":"98","author":"Kobayashi","year":"2000","journal-title":"Mech. Dev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.mcn.2004.04.010","article-title":"The LIM domain-only protein LMO4 is required for neural tube closure","volume":"28","author":"Lee","year":"2005","journal-title":"Mol. Cell. Neurosci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/S0959-4388(02)00287-8","article-title":"Revisiting cell fate specification in the inner ear","volume":"12","author":"Fekete","year":"2002","journal-title":"Curr. Opin. Neurobiol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Hammond, K.L., Loynes, H.E., Mowbray, C., Runke, G., Hammerschmidt, M., Mullins, M.C., Hildreth, V., Chaudhry, B., and Whitfield, T.T. (2009). A late role for bmp2b in the morphogenesis of semicircular canal ducts in the zebrafish inner ear. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0004368"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1002\/dvg.20233","article-title":"Dlx5 andDlx6 homeobox genes are required for specification of the mammalian vestibular apparatus","volume":"44","author":"Robledo","year":"2006","journal-title":"Genes"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1612","DOI":"10.1101\/gad.1303905","article-title":"Wnt-dependent regulation of inner ear morphogenesis is balanced by the opposing and supporting roles of Shh","volume":"19","author":"Riccomagno","year":"2005","journal-title":"Genes Dev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1016\/j.devcel.2004.06.016","article-title":"Hmx2 and Hmx3 Homeobox Genes Direct Development of the Murine Inner Ear and Hypothalamus and Can Be Functionally Replaced by Drosophila Hmx","volume":"7","author":"Wang","year":"2004","journal-title":"Dev. Cell"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1006\/dbio.2002.0713","article-title":"The Dlx5 Homeobox Gene Is Essential for Vestibular Morphogenesis in the Mouse Embryo through a BMP4-Mediated Pathway","volume":"248","author":"Merlo","year":"2002","journal-title":"Dev. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5429","DOI":"10.1523\/JNEUROSCI.2484-17.2018","article-title":"Reciprocal Negative Regulation Between Lmx1a and Lmo4 Is Required for Inner Ear Formation","volume":"38","author":"Huang","year":"2018","journal-title":"J. Neurosci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.yexcr.2019.05.008","article-title":"Inhibition of protein nitration prevents cisplatin-induced inactivation of STAT3 and promotes anti-apoptotic signaling in organ of Corti cells","volume":"381","author":"Rosati","year":"2019","journal-title":"Exp. Cell Res."},{"key":"ref_41","unstructured":"National Center for Biotechnology Information (2019, July 10). ClinVar; Variation ID 442387, Available online: https:\/\/preview.ncbi.nlm.nih.gov\/clinvar\/variation\/442387."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/14\/7\/1371\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:03:02Z","timestamp":1760126582000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/14\/7\/1371"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,28]]},"references-count":41,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2023,7]]}},"alternative-id":["genes14071371"],"URL":"https:\/\/doi.org\/10.3390\/genes14071371","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,28]]}}}