![conference banner](/images/conference-banner.jpeg)
Abstract
Background: Transforming growth factor-β (TGF-β)/nodal signaling is involved in early embryonic patterning in vertebrates. Nodal modulator (Nomo, also called pM5) is a negative regulator of nodal signaling. Currently, the role of nomo gene in cartilage development in vertebrates remains unknown.
Methods: Nomo mutants were generated in a knockout model of zebrafish by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) targeting of the fibronectin type III domain. The expression of related genes, which are critical for chondrogenesis, was analyzed by whole-mount in situ hybridization and qRT-PCR. Whole-mount alcian staining was performed to analyze the cartilage structure.
Results: nomo is highly expressed in various tissues including the cartilage. We successfully constructed a zebrafish nomo knockout model. nomo homozygous mutants exhibited varying degrees of hypoplasia and dysmorphism on 4 and 5 dpf, which is similar to chondrodysplasia in humans. The key genes of cartilage and skeletal development, including sox9a, sox9b, dlx1a, dlx2a, osx, col10a1, and col11a2 were all downregulated in nomo mutants compared with the wildtype.
Conclusion: The nomo gene positively regulates the expression of the master regulator and other key development genes involved in bone formation and cartilage development and it is essential for cartilage development in zebrafish.
Keywords: nomo, CRISPR/Cas9, chondrogenesis, zebrafish, crest cells, in situ.
Current Molecular Medicine
Title:Loss of the Nodal modulator Nomo results in chondrodysplasia in zebrafish
Volume: 18 Issue: 7
Author(s): Linghui Cao, Lingyu Li, Yongqing Li, Jian Zhuang, Yu Chen, Yuequn Wang, Yan Shi, Jimei Chen, Xiaolan Zhu, Yongqi Wan, Fang Li, Wuzhou Yuan, Xiaoyang Mo, Xiangli Ye, Zuoqiong Zhou, Guo Dai, Zhigang Jiang*, Ping Zhu*, Xiushan Wu*Xiongwei Fan*
Affiliation:
- The Center for Heart Development, State Key Lab of Development Biology of Freshwater Fish, The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, Key Lab of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan,China
- Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100,China
- The Center for Heart Development, State Key Lab of Development Biology of Freshwater Fish, The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, Key Lab of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan,China
- The Center for Heart Development, State Key Lab of Development Biology of Freshwater Fish, The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, Key Lab of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan,China
Keywords: nomo, CRISPR/Cas9, chondrogenesis, zebrafish, crest cells, in situ.
Abstract: Background: Transforming growth factor-β (TGF-β)/nodal signaling is involved in early embryonic patterning in vertebrates. Nodal modulator (Nomo, also called pM5) is a negative regulator of nodal signaling. Currently, the role of nomo gene in cartilage development in vertebrates remains unknown.
Methods: Nomo mutants were generated in a knockout model of zebrafish by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) targeting of the fibronectin type III domain. The expression of related genes, which are critical for chondrogenesis, was analyzed by whole-mount in situ hybridization and qRT-PCR. Whole-mount alcian staining was performed to analyze the cartilage structure.
Results: nomo is highly expressed in various tissues including the cartilage. We successfully constructed a zebrafish nomo knockout model. nomo homozygous mutants exhibited varying degrees of hypoplasia and dysmorphism on 4 and 5 dpf, which is similar to chondrodysplasia in humans. The key genes of cartilage and skeletal development, including sox9a, sox9b, dlx1a, dlx2a, osx, col10a1, and col11a2 were all downregulated in nomo mutants compared with the wildtype.
Conclusion: The nomo gene positively regulates the expression of the master regulator and other key development genes involved in bone formation and cartilage development and it is essential for cartilage development in zebrafish.
Export Options
About this article
Cite this article as:
Cao Linghui , Li Lingyu , Li Yongqing , Zhuang Jian , Chen Yu, Wang Yuequn, Shi Yan , Chen Jimei , Zhu Xiaolan , Wan Yongqi, Li Fang , Yuan Wuzhou , Mo Xiaoyang , Ye Xiangli , Zhou Zuoqiong , Dai Guo , Jiang Zhigang *, Zhu Ping *, Wu Xiushan *, Fan Xiongwei *, Loss of the Nodal modulator Nomo results in chondrodysplasia in zebrafish, Current Molecular Medicine 2018; 18 (7) . https://dx.doi.org/10.2174/1566524019666181212095307
DOI https://dx.doi.org/10.2174/1566524019666181212095307 |
Print ISSN 1566-5240 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5666 |
![Wayfinder Image](/images/wayfinder.jpg)
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Nongenomic Actions of Thyroid Hormones: From Basic Research to Clinical Applications. An Update
Immunology, Endocrine & Metabolic Agents in Medicinal Chemistry (Discontinued) Inhibition of Respiration in Yeast by 2-Phenylethylamine
Current Bioactive Compounds Characterization of LC-HCC Fusion Protein of Botulinum Neurotoxin Type A
Protein & Peptide Letters Melatonin and Synthetic Melatonergic Agonists: Actions and Metabolism in the Central Nervous System
Central Nervous System Agents in Medicinal Chemistry Introduction: Strategies, Molecular Targets and Animal Models Useful for Developing Therapies for Alzheimers Disease
Current Medicinal Chemistry - Central Nervous System Agents Molecular Aspects of Melatonin Treatment in Tinnitus: A Review
Current Drug Targets Role of the α7 Nicotinic Acetylcholine Receptor and RIC-3 in the Cholinergic Anti-inflammatory Pathway
Central Nervous System Agents in Medicinal Chemistry Editorial (Thematic Issue: Metabolic Disorders, Drug Development, Drug Design and Biomarkers)
Current Pharmaceutical Design Gene Therapy, A Potential Therapeutic Tool for Neurological and Neuropsychiatric Disorders: Applications, Challenges and Future Perspective
Current Gene Therapy Research Progress of Mitophagy in Alzheimer's Disease
Current Alzheimer Research Developing Antitumor Magnetic Hyperthermia: Principles, Materials and Devices
Recent Patents on Anti-Cancer Drug Discovery Neurotransmitters and Substances of Abuse: Effects on Adult Neurogenesis
Current Neurovascular Research Bioenergetics and Mitochondrial Dysfunction in Aging: Recent Insights for a Therapeutical Approach
Current Pharmaceutical Design Ibogaine in the Treatment of Substance Dependence
Current Drug Abuse Reviews NADPH Oxidase and Neurodegeneration
Current Neuropharmacology Pharmacogenetics and Pharmacotherapy of Military Personnel Suffering from Post-traumatic Stress Disorder
Current Neuropharmacology Effects of the Novel Non-Peptidyl Low Molecular Weight Radical Scavenger IAC in Different Models of Inflammation: A New Perspective in Anti-Inflammatory Therapy
Current Medicinal Chemistry Organoselenium Compounds as Potential Neuroprotective Therapeutic Agents
Current Organic Chemistry Combination Treatment with rt-PA is More Effective than rt-PA Alone in an in Vitro Human Clot Model
Current Neurovascular Research Prevention of Lymphocyte Neurotoxic Effects by microRNA Delivery
MicroRNA