DOCSLIB.ORG

Nogo-A Regulates Myogenesis Via Interacting with Filamin-C

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Nogo-A Regulates Myogenesis Via Interacting with Filamin-C Park et al. Cell Death Discovery (2021) 7:1 https://doi.org/10.1038/s41420-020-00384-x Cell Death Discovery ARTICLE Open Access Nogo-A regulates myogenesis via interacting with Filamin-C SunYoung Park1,2,Ji-HwanPark3, Un-Beom Kang4, Seong-Kyoon Choi5,6,AhmedElfadl1,H.M.ArifUllah1, Myung-Jin Chung1, Ji-Yoon Son1,HyunHoYun1, Jae-Min Park1,Jae-hyukYim1,Seung-JunJung1,Sang-HyupKim1, Young-Chul Choi7, Dae-Seong Kim8, Jin-Hong Shin8, Jin-Sung Park9,KeunHur 10, Sang-Han Lee11,Eun-JooLee1, Daehee Hwang12 and Kyu-Shik Jeong 1,2 Abstract Among the three isoforms encoded by Rtn4, Nogo-A has been intensely investigated as a central nervous system inhibitor. Although Nogo-A expression is increased in muscles of patients with amyotrophic lateral sclerosis, its role in muscle homeostasis and regeneration is not well elucidated. In this study, we discovered a significant increase in Nogo-A expression in various muscle-related pathological conditions. Nogo−/− mice displayed dystrophic muscle structure, dysregulated muscle regeneration following injury, and altered gene expression involving lipid storage and muscle cell differentiation. We hypothesized that increased Nogo-A levels might regulate muscle regeneration. Differentiating myoblasts exhibited Nogo-A upregulation and silencing Nogo-A abrogated myoblast differentiation. Nogo-A interacted with filamin-C, suggesting a role for Nogo-A in cytoskeletal arrangement during myogenesis. In conclusion, Nogo-A maintains muscle homeostasis and integrity, and pathologically altered Nogo-A expression mediates muscle regeneration, suggesting Nogo-A as a novel target for the treatment of myopathies in clinical settings. 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Introduction during myogenesis, a multistep process including trans- Muscular dystrophies are a group of genetic muscle formation of satellite cells into myoblasts, differentiation disorders that cause defective muscle fiber function due to into fusion-competent myoblasts, and their fusion into the weakening and breakdown of muscle fibers as a result multinucleated myotubes5,6. Myogenesis is regulated by of inflammation accompanied with abrogated muscle the synchronized actions of the myogenic regulatory regeneration. Duchenne muscular dystrophy (DMD), the factors myogenic factor 5, MyoD, and myogenin, and most common form of muscular dystrophy, is severe results in the expression of muscle-specific genes7,8. myopathy associated with an abrogated dystrophin gene, During differentiation, the Pax3 and Pax7 expression which results in muscle membrane frailty associated with levels are downregulated, although satellite cells retain – increased pro-inflammatory cytokine levels, mitochon- Pax7 expression during quiescence9 11. – drial dysfunction, and impaired satellite cell polarity1 4. The neurite outgrowth inhibitor Nogo, also known as Following injury, satellite cells transform to committed reticulon 4, is encoded by Rtn4. Independently cloned by – myoblasts and ultimately differentiate into myofibers three groups12 14, Rtn4 complementary DNA (cDNA) was found to generate three Nogo isoforms, Nogo-A, Nogo-B, and Nogo-C, by alternative splicing or distinctive Correspondence: Kyu-Shik Jeong ([email protected]) 15,16 1Department of Pathology, College of Veterinary Medicine, Kyungpook promoter usage . Although mainly localized to the National University, Daegu 41566, Republic of Korea endoplasmic reticulum (ER), Nogo has also been observed 2 Stem Cell Therapeutic Research Institute, Kyungpook National University, on the cell surface17. Nogo-A, the largest Nogo isoform, Daegu 41566, Republic of Korea Full list of author information is available at the end of the article which also carries an inhibitor region named Δ20, is a Edited by Ivano Amelio © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a linktotheCreativeCommons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Official journal of the Cell Death Differentiation Association Park et al. Cell Death Discovery (2021) 7:1 Page 2 of 18 potent neurite growth inhibitor in the central nervous The expression levels of Nogo and myogenic factors are system (CNS)14,15. Nogo-A is expressed in the brain, altered in animal models of muscle disorders spinal cord, eye, and skeletal muscle18, and most of our The muscles of mdx mice, the animal model of human knowledge on Nogo-A concerns its function as a neurite DMD, showed significantly elevated mRNA levels of outgrowth inhibitor in the CNS13,19. On the surface of Nogo-A (Fig. 2A), MyoD, and myogenin (Fig. 2B), and a cells, Nogo-A acts via its receptors Nogo-66 receptor 1, reduced Nogo-C mRNA level (Fig. 2A) compared with the sphingosine-1-phosphate receptor (S1PR) 2, and myelin- wild-type (WT) mice. These results were consistent with associated glycoprotein. Nogo isoforms binds Nogo-66 our analyses in human muscle samples from myopathies receptor 1 and inhibits neurite growth via the Rho/Rho- (Fig. 1B). The observed upregulation in Nogo-A and associated kinase signaling pathway14,18,20,21. However, Myod transcript levels was accompanied by increases in the mechanisms underlying the role of Nogo-A in muscle their protein levels (Fig. 2C, D). Dasarathy et al.22 have regeneration and myogenesis in skeletal muscle remain demonstrated a relationship between skeletal muscle loss elusive. and alcoholic liver disease (ALD); thus, we also analyzed In the present study, we also elucidate the role of Nogo in the skeletal muscle in a mouse model of ALD as a myo- the regulation of muscle homeostasis, muscle differentia- pathy model associated with chronic liver dysfunction. In tion, and adipocyte differentiation based on the muscle current study, the mouse ALD model successfully exhib- + + − − transcriptome profiles of Nogo / and Nogo / mice. We ited liver pathology including hepatomegaly, determined − − use a muscle-injury model in Nogo / mice to examine by the ratio of liver weight to body weight (Fig. S1A), and Nogo-A involvement in muscle regeneration and further elevated levels of plasma aspartate aminotransferase demonstrate the potent myogenic function of Nogo-A in (AST) and alanine aminotransferase (ALT) (Fig. S1C) myoblast differentiation using muscle lineage cells, compared to the control mice. Therefore, the elevated including C2C12 cells and induced muscle stem cells plasma ALT and AST levels in ALD mice suggest liver (iMSCs). Our findings suggest that the expression of Nogo- dysfunction as well as muscle damage (Fig. S1C). The A is altered in the skeletal muscle during pathological skeletal muscle tissues of the ALD mice exhibited upre- processes, and that Nogo-A serves a critical function in gulated Nogo-A and reduced Nogo-C mRNA levels muscle regeneration and maintenance of muscle integrity. compared to the controls (Fig. 2E); the tissues also showed elevated levels of myogenin and Pax7 (Fig. 2F), + Results suggesting increased numbers of committed Pax7 Nogo expression is altered in the muscle tissue of patients satellite cells and myogenin-positive myoblasts. with myopathies Acute muscle injury induced by notexin injection was Muscle tissue samples from patients with different associated with a dramatic increase in Nogo-A and a myopathies, including DMD and inflammatory myopathy, significant reduction in Nogo-C (Fig. 2G), as well as sig- exhibited typical histopathological features of muscle nificant upregulation of Myod and myogenin (Fig. 2H). In + degeneration including damaged myofiber structure, these animals, the number of Pax7 committed satellite inflammatory cell infiltration, and fibrosis resulting from cells was not altered but the MYH2 expression was dra- incomplete muscle regeneration (Fig. 1A). Thus, we matically downregulated, indicating structural muscle examined the transcriptional expression levels of Nogo damage (Fig. 2H). isoforms and myogenic factors in muscle tissues from patients with myopathies including DMD and inflamma- Muscle lacking Nogo exhibits abnormal structure and tory myopathy, which revealed that the levels of Nogo impaired regeneration following notexin-induced muscle isoforms were significantly altered in the muscle samples injury + + of patients with myopathies (Fig. 1B). Specifically, Although the Nogo-A / muscles exhibited well- although the level of Nogo-B in myopathic tissues was structured muscle fibers and the absence of inflamma- − − comparable to that in normal muscle tissues, the level of tory cell infiltration, the Nogo / muscles displayed Nogo-A was dramatically elevated and the level of Nogo- defective myofibers and exhibited immune cell infiltration C was significantly reduced in the myopathic tissues (Fig. 3A, right panel, arrow). Notexin-induced muscle compared with the normal tissues (Fig. 1B). The tran- damage results in inflammation at the site of injection script levels of Pax7,
Load more

Recommended publications
  • Detailed Review Paper on Retinoid Pathway Signalling
    1 1 Detailed Review Paper on Retinoid Pathway Signalling 2 December 2020 3 2 4 Foreword 5 1. Project 4.97 to develop a Detailed Review Paper (DRP) on the Retinoid System 6 was added to the Test Guidelines Programme work plan in 2015. The project was 7 originally proposed by Sweden and the European Commission later joined the project as 8 a co-lead. In 2019, the OECD Secretariat was added to coordinate input from expert 9 consultants. The initial objectives of the project were to: 10 draft a review of the biology of retinoid signalling pathway, 11 describe retinoid-mediated effects on various organ systems, 12 identify relevant retinoid in vitro and ex vivo assays that measure mechanistic 13 effects of chemicals for development, and 14 Identify in vivo endpoints that could be added to existing test guidelines to 15 identify chemical effects on retinoid pathway signalling. 16 2. This DRP is intended to expand the recommendations for the retinoid pathway 17 included in the OECD Detailed Review Paper on the State of the Science on Novel In 18 vitro and In vivo Screening and Testing Methods and Endpoints for Evaluating 19 Endocrine Disruptors (DRP No 178). The retinoid signalling pathway was one of seven 20 endocrine pathways considered to be susceptible to environmental endocrine disruption 21 and for which relevant endpoints could be measured in new or existing OECD Test 22 Guidelines for evaluating endocrine disruption. Due to the complexity of retinoid 23 signalling across multiple organ systems, this effort was foreseen as a multi-step process.
    [Show full text]
  • A Population-Based Study of Effects of Genetic Loci on Orofacial Clefts
    HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author J Dent Res Manuscript Author . Author manuscript; Manuscript Author available in PMC 2017 October 01. Published in final edited form as: J Dent Res. 2017 October ; 96(11): 1322–1329. doi:10.1177/0022034517716914. A Population-Based Study of Effects of Genetic Loci on Orofacial Clefts L.M. Moreno Uribe1, T. Fomina2, R.G. Munger3, P.A. Romitti4, M.M. Jenkins5, H.K. Gjessing2,6, M. Gjerdevik2,6, K. Christensen7, A.J. Wilcox8, J.C. Murray9, R.T. Lie2,6,*, and G.L. Wehby10,* 1Department of Orthodontics and Dows Institute, College of Dentistry, University of Iowa, Iowa City, IA, USA 2Department of Global Public Health and Primary Care, University of Bergen, Bergen, Norway 3Department of Nutrition and Food Sciences, Utah State University, Logan, UT, USA 4Department of Epidemiology, College of Public Health, University of Iowa, Iowa City, IA, USA 5National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta, GA, USA 6Norwegian Institute of Public Health, Bergen and Oslo, Norway 7Department of Public Health, University of Southern Denmark; Department of Clinical Genetics and Department of Biochemistry and Pharmacology, Odense University Hospital, Odense, Denmark 8Epidemiology Branch, National Institute of Environmental Health Sciences, National Institutes of Health, Durham, NC, USA 9Department of Pediatrics, Carver College of Medicine, University of Iowa, Iowa City, IA, USA 10Departments of Health Management and Policy, Economics, and Preventive and Community Dentistry, and Public Policy Center, University of Iowa, Iowa City, IA, USA Abstract Prior genome-wide association studies for oral clefts have focused on clinic-based samples with unclear generalizability.
    [Show full text]
  • Differentiation of the Human PAX7
    © 2020. Published by The Company of Biologists Ltd | Development (2020) 147, dev187344. doi:10.1242/dev.187344 HUMAN DEVELOPMENT TECHNIQUES AND RESOURCES ARTICLE Differentiation of the human PAX7-positive myogenic precursors/satellite cell lineage in vitro Ziad Al Tanoury1,2,3, Jyoti Rao2,3, Olivier Tassy1,Bénédicte Gobert1,4, Svetlana Gapon2, Jean-Marie Garnier1, Erica Wagner2, Aurore Hick4, Arielle Hall5, Emanuela Gussoni5 and Olivier Pourquié1,2,3,6,* ABSTRACT SC derive from the paraxial mesoderm, the embryonic tissue that Satellite cells (SC) are muscle stem cells that can regenerate adult forms the vertebral column and skeletal muscles (Chal and Pourquié, muscles upon injury. Most SC originate from PAX7+ myogenic 2017; Gros et al., 2005; Hutcheson et al., 2009; Kassar-Duchossoy precursors set aside during development. Although myogenesis has et al., 2005; Lepper and Fan, 2010; Relaix et al., 2005; Schienda et al., + been studied in mouse and chicken embryos, little is known about 2006). Pax3 myogenic precursors arise from the dorsal epithelial + human muscle development. Here, we report the generation of human compartment of the somite called the dermomyotome. Pax3 cells of induced pluripotent stem cell (iPSC) reporter lines in which fluorescent the dermomyotome lips activate Myf5, then downregulate Pax3 and proteins have been introduced into the PAX7 and MYOG loci. We use delaminate and differentiate into elongated post-mitotic myocytes single cell RNA sequencing to analyze the developmental trajectory of expressing myogenin (Myog) to form the first embryonic muscles + the iPSC-derived PAX7+ myogenic precursors. We show that the called myotomes (Chal and Pourquié, 2017). At the same time, Pax3 PAX7+ cells generated in culture can produce myofibers and self- myogenic precursors delaminate from the lateral edge of the renew in vitro and in vivo.
    [Show full text]
  • PAX7 Balances the Cell Cycle Progression Via Regulating Expression of Dnmt3b and Apobec2 in Differentiating Pscs
    cells Article PAX7 Balances the Cell Cycle Progression via Regulating Expression of Dnmt3b and Apobec2 in Differentiating PSCs Anita Florkowska , Igor Meszka, Joanna Nowacka, Monika Granica , Zuzanna Jablonska, Magdalena Zawada, Lukasz Truszkowski , Maria A. Ciemerych and Iwona Grabowska * Department of Cytology, Institute of Developmental Biology and Biomedical Sciences, Faculty of Biology, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland; a.fl[email protected] (A.F.); [email protected] (I.M.); [email protected] (J.N.); [email protected] (M.G.); [email protected] (Z.J.); [email protected] (M.Z.); [email protected] (L.T.); [email protected] (M.A.C.) * Correspondence: [email protected] Abstract: PAX7 transcription factor plays a crucial role in embryonic myogenesis and in adult muscles in which it secures proper function of satellite cells, including regulation of their self renewal. PAX7 downregulation is necessary for the myogenic differentiation of satellite cells induced after muscle damage, what is prerequisite step for regeneration. Using differentiating pluripotent stem cells we documented that the absence of functional PAX7 facilitates proliferation. Such action is executed by the modulation of the expression of two proteins involved in the DNA methylation, i.e., Dnmt3b and Apobec2. Increase in Dnmt3b expression led to the downregulation of the CDK inhibitors and Citation: Florkowska, A.; Meszka, I.; facilitated cell cycle progression. Changes in Apobec2 expression, on the other hand, differently Nowacka, J.; Granica, M.; Jablonska, impacted proliferation/differentiation balance, depending on the experimental model used. Z.; Zawada, M.; Truszkowski, L.; Ciemerych, M.A.; Grabowska, I.
    [Show full text]
  • An OTX2-PAX3 Signaling Axis Regulates Group 3 Medulloblastoma Cell Fate
    ARTICLE https://doi.org/10.1038/s41467-020-17357-4 OPEN An OTX2-PAX3 signaling axis regulates Group 3 medulloblastoma cell fate Jamie Zagozewski1, Ghazaleh M. Shahriary 1, Ludivine Coudière Morrison1, Olivier Saulnier 2,3, Margaret Stromecki1, Agnes Fresnoza4, Gareth Palidwor5, Christopher J. Porter 5, Antoine Forget6,7, Olivier Ayrault 6,7, Cynthia Hawkins2,8,9, Jennifer A. Chan10, Maria C. Vladoiu2,3,9, Lakshmikirupa Sundaresan2,3, Janilyn Arsenio11,12, Michael D. Taylor 2,3,9,13, Vijay Ramaswamy 2,3,14,15 & ✉ Tamra E. Werbowetski-Ogilvie 1 1234567890():,; OTX2 is a potent oncogene that promotes tumor growth in Group 3 medulloblastoma. However, the mechanisms by which OTX2 represses neural differentiation are not well characterized. Here, we perform extensive multiomic analyses to identify an OTX2 regulatory network that controls Group 3 medulloblastoma cell fate. OTX2 silencing modulates the repressive chromatin landscape, decreases levels of PRC2 complex genes and increases the expression of neurodevelopmental transcription factors including PAX3 and PAX6. Expression of PAX3 and PAX6 is significantly lower in Group 3 medulloblastoma patients and is cor- related with reduced survival, yet only PAX3 inhibits self-renewal in vitro and increases survival in vivo. Single cell RNA sequencing of Group 3 medulloblastoma tumorspheres demonstrates expression of an undifferentiated progenitor program observed in primary tumors and characterized by translation/elongation factor genes. Identification of mTORC1 signaling as a downstream effector of OTX2-PAX3 reveals roles for protein synthesis pathways in regulating Group 3 medulloblastoma pathogenesis. 1 Regenerative Medicine Program, Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB, Canada. 2 The Arthur and Sonia Labatt Brain Tumour Research Center, The Hospital for Sick Children, Toronto, ON, Canada.
    [Show full text]
  • Effects of Estrogen Receptor Activation on Post-Exercise Muscle Satellite Cell Proliferation
    Wilfrid Laurier University Scholars Commons @ Laurier Theses and Dissertations (Comprehensive) 2009 Effects of Estrogen Receptor Activation on Post-Exercise Muscle Satellite Cell Proliferation Kareem Bunyan Wilfrid Laurier University Follow this and additional works at: https://scholars.wlu.ca/etd Part of the Exercise Science Commons Recommended Citation Bunyan, Kareem, "Effects of Estrogen Receptor Activation on Post-Exercise Muscle Satellite Cell Proliferation" (2009). Theses and Dissertations (Comprehensive). 928. https://scholars.wlu.ca/etd/928 This Thesis is brought to you for free and open access by Scholars Commons @ Laurier. It has been accepted for inclusion in Theses and Dissertations (Comprehensive) by an authorized administrator of Scholars Commons @ Laurier. For more information, please contact [email protected]. Library and Archives Bibliotheque et 1*1 Canada Archives Canada Published Heritage Direction du Branch Patrimoine de I'edition 395 Wellington Street 395, rue Wellington Ottawa ON K1A 0N4 OttawaONK1A0N4 Canada Canada Your file Votre r&ference ISBN: 978-0-494-54223-1 Our file Notre reference ISBN: 978-0-494-54223-1 NOTICE: AVIS: The author has granted a non­ L'auteur a accorde une licence non exclusive exclusive license allowing Library and permettant a la Bibliotheque et Archives Archives Canada to reproduce, Canada de reproduire, publier, archiver, publish, archive, preserve, conserve, sauvegarder, conserver, transmettre au public communicate to the public by par telecommunication ou par I'lnternet, prefer, telecommunication or on the Internet, distribuer et vendre des theses partout dans le loan, distribute and sell theses monde, a des fins commerciales ou autres, sur worldwide, for commercial or non­ support microforme, papier, electronique et/ou commercial purposes, in microform, autres formats.
    [Show full text]
  • A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family
    Rockefeller University Digital Commons @ RU Student Theses and Dissertations 2018 A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family Linda Molla Follow this and additional works at: https://digitalcommons.rockefeller.edu/ student_theses_and_dissertations Part of the Life Sciences Commons A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY A Thesis Presented to the Faculty of The Rockefeller University in Partial Fulfillment of the Requirements for the degree of Doctor of Philosophy by Linda Molla June 2018 © Copyright by Linda Molla 2018 A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY Linda Molla, Ph.D. The Rockefeller University 2018 APOBEC2 is a member of the AID/APOBEC cytidine deaminase family of proteins. Unlike most of AID/APOBEC, however, APOBEC2’s function remains elusive. Previous research has implicated APOBEC2 in diverse organisms and cellular processes such as muscle biology (in Mus musculus), regeneration (in Danio rerio), and development (in Xenopus laevis). APOBEC2 has also been implicated in cancer. However the enzymatic activity, substrate or physiological target(s) of APOBEC2 are unknown. For this thesis, I have combined Next Generation Sequencing (NGS) techniques with state-of-the-art molecular biology to determine the physiological targets of APOBEC2. Using a cell culture muscle differentiation system, and RNA sequencing (RNA-Seq) by polyA capture, I demonstrated that unlike the AID/APOBEC family member APOBEC1, APOBEC2 is not an RNA editor. Using the same system combined with enhanced Reduced Representation Bisulfite Sequencing (eRRBS) analyses I showed that, unlike the AID/APOBEC family member AID, APOBEC2 does not act as a 5-methyl-C deaminase.
    [Show full text]
  • Identification of Shared and Unique Gene Families Associated with Oral
    International Journal of Oral Science (2017) 9, 104–109 OPEN www.nature.com/ijos ORIGINAL ARTICLE Identification of shared and unique gene families associated with oral clefts Noriko Funato and Masataka Nakamura Oral clefts, the most frequent congenital birth defects in humans, are multifactorial disorders caused by genetic and environmental factors. Epidemiological studies point to different etiologies underlying the oral cleft phenotypes, cleft lip (CL), CL and/or palate (CL/P) and cleft palate (CP). More than 350 genes have syndromic and/or nonsyndromic oral cleft associations in humans. Although genes related to genetic disorders associated with oral cleft phenotypes are known, a gap between detecting these associations and interpretation of their biological importance has remained. Here, using a gene ontology analysis approach, we grouped these candidate genes on the basis of different functional categories to gain insight into the genetic etiology of oral clefts. We identified different genetic profiles and found correlations between the functions of gene products and oral cleft phenotypes. Our results indicate inherent differences in the genetic etiologies that underlie oral cleft phenotypes and support epidemiological evidence that genes associated with CL/P are both developmentally and genetically different from CP only, incomplete CP, and submucous CP. The epidemiological differences among cleft phenotypes may reflect differences in the underlying genetic causes. Understanding the different causative etiologies of oral clefts is
    [Show full text]
  • Pioneer Factor Pax7 Deploys a Stable Enhancer Repertoire for Specification of Cell Fate
    ARTICLES https://doi.org/10.1038/s41588-017-0035-2 Pioneer factor Pax7 deploys a stable enhancer repertoire for specification of cell fate Alexandre Mayran 1,2, Konstantin Khetchoumian1, Fadi Hariri3, Tomi Pastinen3, Yves Gauthier1, Aurelio Balsalobre1 and Jacques Drouin1,2* Pioneer transcription factors establish new cell-fate competence by triggering chromatin remodeling. However, many features of pioneer action, such as their kinetics and stability, remain poorly defined. Here, we show that Pax7, by opening a unique repertoire of enhancers, is necessary and sufficient for specification of one pituitary lineage. Pax7 binds its targeted enhancers rapidly, but chromatin remodeling and gene activation are slower. Enhancers opened by Pax7 show a loss of DNA methylation and acquire stable epigenetic memory, as evidenced by binding of nonpioneer factors after Pax7 withdrawal. This work shows that transient Pax7 expression is sufficient for stable specification of cell identity. ell specification and differentiation occur throughout devel- DNA target sites in heterochromatin; (ii) initiate remodeling of the opment in multicellular organisms and, in complex life forms, surrounding chromatin; (iii) facilitate binding of other TFs; and (iv) Clead to a high diversity of cells. The differentiation of cells trigger stable changes in chromatin accessibility, thus ensuring epi- into different lineages is implemented by a unique combination of genetic stability, i.e., long-term access by nonpioneer factors. transcription factors (TFs) that collectively control the program of We now show that the bulk of differentially accessible chromatin cell-specific gene expression. During development, progenitors are regions between the two alternate POMC cell fates are at putative specified by the action of selector genes that establish the differen- enhancers.
    [Show full text]
  • Pax3 and Pax7 Exhibit Distinct and Overlapping Functions in Marking Muscle Satellite Cells and Muscle Repair in a Marine Teleost, Sebastes Schlegelii
    International Journal of Molecular Sciences Article Pax3 and Pax7 Exhibit Distinct and Overlapping Functions in Marking Muscle Satellite Cells and Muscle Repair in a Marine Teleost, Sebastes schlegelii Mengya Wang 1,2, Weihao Song 1 , Chaofan Jin 1, Kejia Huang 1, Qianwen Yu 1, Jie Qi 1,2, Quanqi Zhang 1,2 and Yan He 1,2,* 1 MOE Key Laboratory of Molecular Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China; [email protected] (M.W.); [email protected] (W.S.); [email protected] (C.J.); [email protected] (K.H.); [email protected] (Q.Y.); [email protected] (J.Q.); [email protected] (Q.Z.) 2 Laboratory of Tropical Marine Germplasm Resources and Breeding Engineering, Sanya Oceanographic Institution, Ocean University of China, Sanya 572000, China * Correspondence: [email protected] Abstract: Pax3 and Pax7 are members of the Pax gene family which are essential for embryo and organ development. Both genes have been proved to be markers of muscle satellite cells and play key roles in the process of muscle growth and repair. Here, we identified two Pax3 genes (SsPax3a and SsPax3b) and two Pax7 genes (SsPax7a and SsPax7b) in a marine teleost, black rockfish (Sebastes schlegelii). Our results showed SsPax3 and SsPax7 marked distinct populations of muscle satellite cells, which originated from the multi-cell stage and somite stage, respectively. In addition, we Citation: Wang, M.; Song, W.; Jin, C.; constructed a muscle injury model to explore the function of these four genes during muscle repair. Huang, K.; Yu, Q.; Qi, J.; Zhang, Q.; Hematoxylin–eosin (H–E) of injured muscle sections showed new-formed myofibers occurred at He, Y.
    [Show full text]
  • M. Garcia-Castro
    MIGCrest lab Examining the State of the Science of Mammalian Embryo Model Systems: A Workshop (NAS) Jan 17th 2020 SESSION IV: COMPARATIVE EMBRYONIC DEVELOPMENT ACROSS SPECIES Early Neural Crest Formation: from birds to humans Division of Biomedical SciencesGarcía-Castro Lab UCR, School of Medicine Wilhelm His, 1868 Chick embryo 36 hours dev Head Tail Zwischengstrang, Zwischenrinne Middle cord, Marshall 1879… “NEURAL CREST” Chick embryo Head Neural Crest Migrate extensively through stereotypic patterns Tail Chick embryo Head Neural Crest Derivatives Cartilage, bone, and Connective tissue neurons & glia of the PNS, and Melanocytes Sympathoadrenal cells PATHOLOGIES: 1/3 of all birth defects. Tail Orofacial clefts, rare syndromes, and cancers MIGCrest lab Differentiation Potential of Neural Crest Neurons, Glia, melanocytes, Adipocytes, odontoblast, osteoblasts, chondroblasts, myocytes, Oxigen sensing cells of the Carotid Body, Thymus mesenchyme, etc.… How do NC cells Generate so Many Different Cell types? EPIBLAST ESC Gastrulation Ectoderm Mesoderm Endoderm Skin & CNS Muscle, Bone, Adipo, Blood Gut, Thyroid, Lungs, Pancreas EPIBLAST Melanocytes PNS Gastrulation Neural Crest Cells Ectoderm Mesoderm Endoderm Skin & CNS Muscle, Bone, Adipo, Blood Gut, Thyroid, Lungs, Pancreas Waddington Epigenetics Sequential segregation of potential CRANIOFACIAL Bone , Cartilage, EPIBLAST Muscle, Odontoblast, Revisit the OntogenyAdipocytes, Gastrulation Connective of the Neural Cresttissue, etc. Ectoderm Mesoderm Heart valves Skin & CNS Muscle, Bone, Adipo, Blood Garcia-Castro Lab: Neural Crest Ontogeny Pax3/7 KO and Cre-lines/ROSA26 Rabbit MIGCrest lab ”Ontogeny of Neural Crest " Specification of chick NC before gastrulation. Human NC, Embryonic studies and PSC models suggest an early post epiblast differentiation. Specification of mammalian NC in the Rabbit is evident during gastrulation García-Castro Lab UCR, School of Medicine Neural Crest Formation Classic Induction Wnt BMP Cascade of FGF Transcription Factors and other markers responsible for Neural Crest dev.
    [Show full text]
  • Quantitative Analyses of the Global Proteome and Phosphoproteome Reveal the Different Impacts of Propofol and Dexmedetomidine on HT22 Cells
    www.nature.com/scientificreports OPEN Quantitative analyses of the global proteome and phosphoproteome reveal the different impacts of Received: 21 July 2016 Accepted: 17 March 2017 propofol and dexmedetomidine on Published: 18 April 2017 HT22 cells Honggang Zhang1, Juan Ye2, Zhaomei Shi3, Chen Bu3 & Fangping Bao1 Propofol and dexmedetomidine are both commonly used anaesthetics. Although they employ two different mechanisms to induce anaesthesia, both compounds influence the hippocampus and the HT22 cell line. HT22 cells are broadly used in neurobiological research. In this study, we assessed the effects of propofol and dexmedetomidine on signalling in HT22 cells. Using the SILAC (stable isotope labelling with amino acids in cell culture) labelling technique, IMAC (immobilized metal affinity chromatography) enrichment and high-resolution LC-MS/MS (liquid chromatography tandem mass spectrometry) analysis, we investigated the quantitative proteome and phosphoproteome in HT22 cells treated with propofol or dexmedetomidine. In total, 4,527 proteins and 6,824 phosphosites were quantified in cells treated with these two anaesthetics. With the assistance of intensive bioinformatics, the propofol and dexmedetomidine treatments were shown to induce distinct proteome and phosphoproteome profiles in HT22 cells. Consistent with our bioinformatics analysis, dexmedetomidine had a smaller effect than propofol on cell survival. These findings deepen our understanding of drug-induced anaesthesia. Not surprisingly, the development of anaesthesia has substantially contributed to modern surgery. However, the potential side effects of anaesthetics and the increasing demand for better medical services have prompted researchers to obtain a deeper understanding of the mechanisms of commonly used anaesthetics1. Due to its antioxidant, anti-inflammatory and bronchiectasis properties2, propofol (2,6-diisopropylphenol) has been widely used to induce and maintain general anaesthesia since its discovery in 19773.
    [Show full text]