DOCSLIB.ORG

Ebf2 Is a Selective Marker of Brown and Beige Adipogenic Precursor Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Ebf2 Is a Selective Marker of Brown and Beige Adipogenic Precursor Cells Ebf2 is a selective marker of brown and beige adipogenic precursor cells Wenshan Wanga,b, Megan Kissiga,b, Sona Rajakumaria,b, Li Huanga,b, Hee-woong Lima,c, Kyoung-Jae Wona,c, and Patrick Sealea,b,1 aInstitute for Diabetes, Obesity, Metabolism, bDepartment of Cell and Developmental Biology, and cDepartment of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104 Edited by C. Ronald Kahn, Joslin Diabetes Center, Harvard Medical School, Boston, MA, and approved August 8, 2014 (received for review July 8, 2014) Brown adipocytes and muscle and dorsal dermis descend from transcriptomic analyses identified a brown-preadipose–specific precursor cells in the dermomyotome, but the factors that regulate gene signature, which included early B-cell factor 2 (Ebf2), a crit- commitment to the brown adipose lineage are unknown. Here, we ical transcriptional regulator in mature brown adipocytes (18). prospectively isolated and determined the molecular profile of We found that brown/beige preadipose activity in embryos and embryonic brown preadipose cells. Brown adipogenic precursor adult fat depots was restricted to Ebf2-expressing cells. Further- activity in embryos was confined to platelet-derived growth factor more, Ebf2 deficiency in brown preadipose cells reduced the ex- + Cre α , myogenic factor 5 -lineage–marked cells. RNA-sequence anal- pression of nearly all brown preadipose signature genes, whereas ysis identified early B-cell factor 2 (Ebf2) as one of the most selec- ectopic expression of Ebf2 in myoblasts activated brown pre- tively expressed genes in this cell fraction. Importantly, Ebf2- GFP adipose-selective genes. Altogether, our study reveals that expressing cells purified from Ebf2 embryos or brown fat tissue Ebf2 is a specific marker of brown/beige preadipose cells and did not express myoblast or dermal cell markers and uniformly dif- that Ebf2 functions at this stage to control precursor identity. ferentiated into brown adipocytes. Interestingly, Ebf2-expressing cells from white fat tissue in adult animals differentiated into Results brown-like (or beige) adipocytes. Loss of Ebf2 in brown preadipose Prospective Isolation of Myf5Cre-Lineage–Marked Brown Preadipose cells reduced the expression levels of brown preadipose-signature Cells. Specific marker genes for brown preadipose cells were pre- genes, whereas ectopic Ebf2 expression in myoblasts activated viously unknown. To identify such factors, we isolated enriched brown preadipose-specific genes. Altogether, these results indicate populations of brown preadipose cells from mouse embryos. that Ebf2 specifically marks and regulates the molecular profile of Brown adipocytes descend from a Myf5Cre-expressing cell lineage brown preadipose cells. that also gives rise to skeletal muscle cells (17). To enable the Cre – beige adipocyte | brown adipose tissue efficient purification of Myf5 -lineage marked cells by flow cytometry, we generated Myf5Cre; R26R-mTmG mice, in which Myf5Cre-expressing cells are genetically and heritably labeled by rown adipose tissue (BAT) expends energy in the form of a membrane-targeted form of GFP [Myf5Cre(GFP)] (19). We heat in response to various stimuli, including cold exposure B noted that GFP was readily detected in somites starting from and certain diets (1). The thermogenic activity of brown adipo- embryonic day (E) ∼E9.5 (Fig. S1A). In agreement with previous cytes is mediated by uncoupling protein 1 (Ucp1), a mitochondrial findings, the majority of peroxisome proliferator-activated receptor membrane protein. Ucp1, when activated, dissipates the electro- chemical gradient that powers ATP synthesis, which allows for low rates of ATP production despite high levels of substrate oxidation Significance (1). Thermogenically competent Ucp1-expressing brown-like adi- pocytes, called beige or brite (brown-in-white) cells, also develop High levels of brown/beige fat activity protects animals against in certain white adipose tissue (WAT) depots in response to cold metabolic disease, but there has been little known about the exposure (reviewed in ref. 2). In mice, BAT and/or beige fat- precursor cells that mediate the expansion of brown or beige mediated thermogenesis suppresses high-fat-diet–associated obesity fat. We discovered that early B-cell factor 2 (Ebf2), a transcrip- – tion factor, is selectively expressed in brown and beige fat cell and metabolic disease (2 8). High levels of activated BAT also + correlate with reduced adiposity in people (9). There is thus great precursors. Through purification of Ebf2 cells, we identified hope that brown/beige fat activity can be increased to reduce obesity a gene profile of brown fat precursors that can be used to and metabolic disease. distinguish these cells from other developmentally related cell BAT depots mature during the embryonic and fetal stages of types. Importantly, Ebf2 was also found to regulate the gene mouse development. In humans, the major BAT depot is lo- expression profile of brown fat precursor cells. Taken together, cated in the interscapular region of newborns, but this tissue this study identifies Ebf2 as a highly specific marker of brown regresses and is absent in adults (10). However, brown and and beige preadipose cells and reveals that Ebf2 functions to beige adipocytes are found in other depots of adult humans, control brown preadipose cell identity. particularly in the supraclavicular area, neck, and along the – Author contributions: W.W. and P.S. designed research; W.W., M.K., S.R., L.H., and H.-w.L. spinal column (10 14). Recent lineage studies showed that performed research; S.R. contributed new reagents/analytic tools; W.W., H.-w.L., K.-J.W., brown adipocytes originate from precursor cells that express and P.S. analyzed data; and W.W. and P.S. wrote the paper. myogenic factor 5 (Myf5), paired box protein 7 (Pax7), Pax3,and The authors declare no conflict of interest. engrailed 1 (En1) in the somitic mesoderm (15–17). However, “ This article is a PNAS Direct Submission. the factors that control brown preadipose cell fate in this tri- Data deposition: The sequences reported in this paper have been deposited in the Gene potent” compartment are unknown. Expression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. In the absence of molecular markers for committed brown pre- GSE60443). adipocytes, such cells can only be recognized retrospectively after See Commentary on page 14318. + their differentiation into Ucp1 adipocytes. Here, we com- 1To whom correspondence should be addressed. Email: [email protected]. bined lineage and cell surface markers to prospectively isolate This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. brown preadipose cells during embryogenesis. Global mRNA 1073/pnas.1412685111/-/DCSupplemental. 14466–14471 | PNAS | October 7, 2014 | vol. 111 | no. 40 www.pnas.org/cgi/doi/10.1073/pnas.1412685111 Downloaded by guest on October 1, 2021 γ (Pparγ)-expressing adipocytes in the interscapular BAT as well as E14.5 Ctrl GFP- GFP+ A Cre 104 B Myf5 ;mTmG SEE COMMENTARY the Desmin-expressing skeletal myofibers in the trunk and limb 103 Pdgfrα- Pdgfrα+ Itga7+ Pdgfrα+ + 102 101 were GFP at E16.5 (Fig. S1B) (17). GFP was also expressed in GFP- GFP+ 101 102 103 adipocytes within the axillary and cervical BAT (Fig. S1C)andin the majority of preadipocytes from postnatal BAT (Fig. S1D). Pdgfrα- Pdgfrα+ + GFP cells also contributed to the dorsal dermis (Fig. S1B). These Itga7- Itga7+ Cre GFP+;Pdgfrα- data indicate that Myf5 cells contribute to at least three different 105 45±2.4 20.1±2.5 104 Oil-red-O staining α mesodermal tissue types in late stage embryos: BAT, skeletal α 104 103 103 102 Pdgfr muscle, and dorsal dermis. Consistent with recent studies, GFP was Pdgfr 102 101 17.8±1.5 17.2±1.6 5.4±0.7 also found in some adipocytes in the retroperitoneal and back s.c. 101 102 103 104 101 102 103 Cre Phase WAT of adult mice (20) (Fig. S1E). Thus, Myf5Cre (GFP) is Myf5 (GFP) Itga7 C General adipocyte genes Brown fat-selective genes a selective but not specific marker of developing brown fat cells. Pparγ AdipoQ Ucp1 Cidea Pparα Prdm16 250 700 14 BAT forms prenatally in mice, but the embryonic stages as- ** 12 ** 200 600 sociated with the morphological and molecular differentiation of 500 10 150 400 ** 8 brown adipocytes were unclear. To assess this differentiation, we 100 300 6 200 4 ** 50 performed hematoxylin and eosin (H&E) staining and immu- 100 2 relative mRNA 0 0 0 - + + + - + + - - + + + - + + - + – + α α α α α α α α α α α α α α α α α nohistochemistry on transverse sections of E13.5 E16.5 embryos. α P P P P Ia7+ P P Ia7+ P Ia7+ P P P Ia7+ P P P P Ia7+ P Ia7+ P P + P ba cell ba cell ba cell ba cell ba cell ba cell Cre 3T3-L1 3T3-L1 3T3-L1 3T3-L1 3T3-L1 At E14, we detected distinctive clusters of Myf5 (GFP) pre- GFP - + - + 3T3-L1 GFP - + - + GFP - + - + sumptive brown adipogenic cells that expressed Pparγ (Fig. S2). D GFP+;Itga7+ GFP-; Pdgfrα+ GFP+;Pdgfrα+ Consistent with previous studies, BAT was easily recognized by GFP Pparγ Merge GFP Pparγ Merge GFP Pparγ Merge H&E staining at E15.5 (21, 22) and contained cells that expressed γ the mature adipocyte marker perilipin (Fig. S2). This suggested that Ppar the dorsal interscapular region of E14 embryos is a rich source of pioneering brown adipose precursors. Prdm16 We next considered whether particular cell surface markers could GFP Prdm16 Merge GFP Prdm16 Merge GFP Prdm16 Merge be combined with Myf5Cre-lineage marking to provide additional + selectivity for brown preadipose cells. Platelet-derived growth factor Fig. 1. Myf5Cre-lineage–derived brown adipogenic precursors are Pdgfrα .(A) α α Cells from E14.5 Myf5Cre;mTmGembryos were fractionated based on ex- (Pdgfr ) is a cell surface marker of adipogenic precursors in adult α WAT and muscle (23–26); this prompted us to examine Pdgfrα pression of GFP, Pdgfr , and Itga7.
Load more

Recommended publications
  • Replication of FTO Gene Associated with Lean Mass in a Meta
    www.nature.com/scientificreports OPEN Replication of FTO Gene associated with lean mass in a Meta-Analysis of Genome-Wide Association Studies Shu Ran1, Zi-Xuan Jiang1, Xiao He1, Yu Liu1, Yu-Xue Zhang1, Lei Zhang2,3, Yu-Fang Pei3,4, Meng Zhang5, Rong Hai6, Gui-Shan Gu7, Bao-Lin Liu1, Qing Tian8, Yong-Hong Zhang3,4, Jing-Yu Wang7 & Hong-Wen Deng8* Sarcopenia is characterized by low skeletal muscle, a complex trait with high heritability. With the dramatically increasing prevalence of obesity, obesity and sarcopenia occur simultaneously, a condition known as sarcopenic obesity. Fat mass and obesity-associated (FTO) gene is a candidate gene of obesity. To identify associations between lean mass and FTO gene, we performed a genome-wide association study (GWAS) of lean mass index (LMI) in 2207 unrelated Caucasian subjects and replicated major fndings in two replication samples including 6,004 unrelated Caucasian and 38,292 unrelated Caucasian. We found 29 single nucleotide polymorphisms (SNPs) in FTO signifcantly associated with sarcopenia (combined p-values ranging from 5.92 × 10−12 to 1.69 × 10−9). Potential biological functions of SNPs were analyzed by HaploReg v4.1, RegulomeDB, GTEx, IMPC and STRING. Our results provide suggestive evidence that FTO gene is associated with lean mass. Sarcopenia is a complex disease described as the age-associated loss of skeletal muscle mass, strength and func- tion impairment1,2. Te low skeletal muscle mass will lead to many public health problems such as sarcopenia, osteoporosis and increased mortality3,4, especially in the elderly. Skeletal muscle is heritable with heritability esti- mates of 30–85% for muscle strength and 45–90% for muscle mass5.
    [Show full text]
  • Table S1 the Four Gene Sets Derived from Gene Expression Profiles of Escs and Differentiated Cells
    Table S1 The four gene sets derived from gene expression profiles of ESCs and differentiated cells Uniform High Uniform Low ES Up ES Down EntrezID GeneSymbol EntrezID GeneSymbol EntrezID GeneSymbol EntrezID GeneSymbol 269261 Rpl12 11354 Abpa 68239 Krt42 15132 Hbb-bh1 67891 Rpl4 11537 Cfd 26380 Esrrb 15126 Hba-x 55949 Eef1b2 11698 Ambn 73703 Dppa2 15111 Hand2 18148 Npm1 11730 Ang3 67374 Jam2 65255 Asb4 67427 Rps20 11731 Ang2 22702 Zfp42 17292 Mesp1 15481 Hspa8 11807 Apoa2 58865 Tdh 19737 Rgs5 100041686 LOC100041686 11814 Apoc3 26388 Ifi202b 225518 Prdm6 11983 Atpif1 11945 Atp4b 11614 Nr0b1 20378 Frzb 19241 Tmsb4x 12007 Azgp1 76815 Calcoco2 12767 Cxcr4 20116 Rps8 12044 Bcl2a1a 219132 D14Ertd668e 103889 Hoxb2 20103 Rps5 12047 Bcl2a1d 381411 Gm1967 17701 Msx1 14694 Gnb2l1 12049 Bcl2l10 20899 Stra8 23796 Aplnr 19941 Rpl26 12096 Bglap1 78625 1700061G19Rik 12627 Cfc1 12070 Ngfrap1 12097 Bglap2 21816 Tgm1 12622 Cer1 19989 Rpl7 12267 C3ar1 67405 Nts 21385 Tbx2 19896 Rpl10a 12279 C9 435337 EG435337 56720 Tdo2 20044 Rps14 12391 Cav3 545913 Zscan4d 16869 Lhx1 19175 Psmb6 12409 Cbr2 244448 Triml1 22253 Unc5c 22627 Ywhae 12477 Ctla4 69134 2200001I15Rik 14174 Fgf3 19951 Rpl32 12523 Cd84 66065 Hsd17b14 16542 Kdr 66152 1110020P15Rik 12524 Cd86 81879 Tcfcp2l1 15122 Hba-a1 66489 Rpl35 12640 Cga 17907 Mylpf 15414 Hoxb6 15519 Hsp90aa1 12642 Ch25h 26424 Nr5a2 210530 Leprel1 66483 Rpl36al 12655 Chi3l3 83560 Tex14 12338 Capn6 27370 Rps26 12796 Camp 17450 Morc1 20671 Sox17 66576 Uqcrh 12869 Cox8b 79455 Pdcl2 20613 Snai1 22154 Tubb5 12959 Cryba4 231821 Centa1 17897
    [Show full text]
  • An Enhancer Directs Differential Expression of the Linked Mrf4 and Myf5 Myogenic Regulatory Genes in the Mouse
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Developmental Biology 269 (2004) 595–608 www.elsevier.com/locate/ydbio An enhancer directs differential expression of the linked Mrf4 and Myf5 myogenic regulatory genes in the mouse Ted Hung-Tse Chang, Michael Primig,1 Juliette Hadchouel,2 Shahragim Tajbakhsh,3 Didier Rocancourt, Anne Fernandez,4 Roland Kappler,5 Harry Scherthan,6 and Margaret Buckingham* De´partement de Biologie du De´veloppement, CNRS URA 2578, Institut Pasteur, 75724 Paris Cedex 15, France Received for publication 25 September 2003, revised 3 February 2004, accepted 6 February 2004 Abstract The myogenic regulatory factors, Mrf4 and Myf5, play a key role in skeletal muscle formation. An enhancer trap approach, devised to isolate positive-acting elements from a 200-kb YAC covering the mouse Mrf4–Myf5 locus in a C2 myoblast assay, yielded an enhancer, A17, which mapped at À8kb5V of Mrf4 and À17 kb 5V of Myf5. An E-box bound by complexes containing the USF transcription factor is critical for enhancer activity. In transgenic mice, A17 gave two distinct and mutually exclusive expression profiles before birth, which correspond to two phases of Mrf4 transcription. Linked to the Tk or Mrf4 minimal promoters, the nlacZ reporter was expressed either in embryonic myotomes, or later in fetal muscle, with the majority of Mrf4 lines showing embryonic expression. When linked to the Myf5 minimal promoter, only fetal muscle expression was detected. These observations identify A17 as a sequence that targets sites of myogenesis in vivo and raise questions about the mutually exclusive modes of expression and possible promoter/enhancer interactions at the Mrf4–Myf5 locus.
    [Show full text]
  • Roles of Twist1 and Twist2 As Molecular Switches During Gene Transcription Hector L
    Published online 8 October 2010 Nucleic Acids Research, 2011, Vol. 39, No. 4 1177–1186 doi:10.1093/nar/gkq890 SURVEY AND SUMMARY Redundant or separate entities?—roles of Twist1 and Twist2 as molecular switches during gene transcription Hector L. Franco, Jose´ Casasnovas, Jose´ R. Rodrı´guez-Medina and Carmen L. Cadilla* Human Molecular Genetics Lab, Department of Biochemistry, School of Medicine University of Puerto Rico, Medical Sciences Campus, PO Box 365067, San Juan, PR 00936, USA Received December 28, 2009; Revised September 8, 2010; Accepted September 19, 2010 ABSTRACT INTRODUCTION Twist1 and Twist2 are highly conserved members of The basic helix–loop–helix (bHLH) family of proteins the Twist subfamily of bHLH proteins responsible comprises a series of transcription factors that act as for the transcriptional regulation of the develop- master regulators on different tissues. Their complex regu- mental programs in mesenchymal cell lineages. latory functions make them fine-tuned machinery for controlling cell fate at the early stages of embryogenesis. The regulation of such processes requires that The hallmark for DNA binding of bHLH transcription Twist1 and Twist2 function as molecular switches factors is the formation of a bipartite DNA-binding to activate and repress target genes by employing domain created when two of these factors form homo or several direct and indirect mechanisms. Modes heterodimer complexes through their HLH motifs of action by these proteins include direct DNA allowing the basic stretch of amino acids to contact the binding to conserved E-box sequences and re- DNA. This DNA-binding domain is able to recognize cis cruitment of coactivators or repressors, seques- regulatory elements containing the consensus sequence tration of E-protein modulators, and interruption 50-NCANNTGN-30 (termed E-box).
    [Show full text]
  • Supplementary Materials
    Supplementary Materials + - NUMB E2F2 PCBP2 CDKN1B MTOR AKT3 HOXA9 HNRNPA1 HNRNPA2B1 HNRNPA2B1 HNRNPK HNRNPA3 PCBP2 AICDA FLT3 SLAMF1 BIC CD34 TAL1 SPI1 GATA1 CD48 PIK3CG RUNX1 PIK3CD SLAMF1 CDKN2B CDKN2A CD34 RUNX1 E2F3 KMT2A RUNX1 T MIXL1 +++ +++ ++++ ++++ +++ 0 0 0 0 hematopoietic potential H1 H1 PB7 PB6 PB6 PB6.1 PB6.1 PB12.1 PB12.1 Figure S1. Unsupervised hierarchical clustering of hPSC-derived EBs according to the mRNA expression of hematopoietic lineage genes (microarray analysis). Hematopoietic-competent cells (H1, PB6.1, PB7) were separated from hematopoietic-deficient ones (PB6, PB12.1). In this experiment, all hPSCs were tested in duplicate, except PB7. Genes under-expressed or over-expressed in blood-deficient hPSCs are indicated in blue and red respectively (related to Table S1). 1 C) Mesoderm B) Endoderm + - KDR HAND1 GATA6 MEF2C DKK1 MSX1 GATA4 WNT3A GATA4 COL2A1 HNF1B ZFPM2 A) Ectoderm GATA4 GATA4 GSC GATA4 T ISL1 NCAM1 FOXH1 NCAM1 MESP1 CER1 WNT3A MIXL1 GATA4 PAX6 CDX2 T PAX6 SOX17 HBB NES GATA6 WT1 SOX1 FN1 ACTC1 ZIC1 FOXA2 MYF5 ZIC1 CXCR4 TBX5 PAX6 NCAM1 TBX20 PAX6 KRT18 DDX4 TUBB3 EPCAM TBX5 SOX2 KRT18 NKX2-5 NES AFP COL1A1 +++ +++ 0 0 0 0 ++++ +++ ++++ +++ +++ ++++ +++ ++++ 0 0 0 0 +++ +++ ++++ +++ ++++ 0 0 0 0 hematopoietic potential H1 H1 H1 H1 H1 H1 PB6 PB6 PB7 PB7 PB6 PB6 PB7 PB6 PB6 PB6.1 PB6.1 PB6.1 PB6.1 PB6.1 PB6.1 PB12.1 PB12.1 PB12.1 PB12.1 PB12.1 PB12.1 Figure S2. Unsupervised hierarchical clustering of hPSC-derived EBs according to the mRNA expression of germ layer differentiation genes (microarray analysis) Selected ectoderm (A), endoderm (B) and mesoderm (C) related genes differentially expressed between hematopoietic-competent (H1, PB6.1, PB7) and -deficient cells (PB6, PB12.1) are shown (related to Table S1).
    [Show full text]
  • Supplementary Materials
    Supplementary Materials Supplementary Note 1 tional clusters with their own provisional cell-type assign- ments. Three of the 10x-only cell types were expected to Inferring cell types, states, and stages be absent from C1 data due to platform-specific details of cell-size filtration (early and late erythrocytes) and sample Differentially expressed genes for each cell cluster were cal- preparation (prior removal of sticky epidermal cells). The culated separately for C1 and 10X datasets using Seurat's remaining 10x only clusters include rarer cell types/states FindMarkers (https://satijalab.org/seurat/seurat_ with no C1 equivalent, presumably due mainly to sparse clustering_tutorial_part2.html) with min.pct = 0.25, cell sampling with C1 plus some additional subdivision of applying its Wilcoxon rank sum test, and setting C1 types into multiple 10x types. min.diff.pct to be 0.2 or 0.4. The differential gene lists are provided in full in Sup- Marker gene sensitivity. For several of the best- plementary Table 4, and representative top markers for studied cell types we found that the C1 data identified key each cluster are given in Tables SN1-1 and SN1-2. The known markers from the literature that were not in the cor- top marker genes for each cell cluster are shown in Fig. 3c responding 10x differential lists. Examples that are func- where each cell cluster was down-sampled to at most 100 tionally important include Pthlh, a regulator of the prolifer- 2,3 cells for 10x data and at most 50 cells for C1 data. ation/differentiation choice in early chondrocytes
    [Show full text]
  • Early Activation of Rat Skeletal Muscle IL-6/STAT1/STAT3 Dependent Gene
    Early activation of rat skeletal muscle IL-6/STAT1/STAT3 dependent gene expression in resistance exercise linked to hypertrophy Gwénaëlle Begue, Aymeric Douillard, Olivier Galbes, Bernadette Rossano, Barbara Vernus, Robin Candau, Guillaume Py To cite this version: Gwénaëlle Begue, Aymeric Douillard, Olivier Galbes, Bernadette Rossano, Barbara Vernus, et al.. Early activation of rat skeletal muscle IL-6/STAT1/STAT3 dependent gene expression in resistance exercise linked to hypertrophy. PLoS ONE, Public Library of Science, 2013, 8 (2), 10.1371/jour- nal.pone.0057141. hal-01837558 HAL Id: hal-01837558 https://hal.archives-ouvertes.fr/hal-01837558 Submitted on 29 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Early Activation of Rat Skeletal Muscle IL-6/STAT1/STAT3 Dependent Gene Expression in Resistance Exercise Linked to Hypertrophy Gwe´nae¨lle Begue, Aymeric Douillard, Olivier Galbes, Bernadette Rossano, Barbara Vernus, Robin Candau, Guillaume Py* INRA, UMR866 Dynamique Musculaire et Me´tabolisme, Universite´ Montpellier 1, F-34060, Montpellier, France Abstract Cytokine interleukin-6 (IL-6) is an essential regulator of satellite cell-mediated hypertrophic muscle growth through the transcription factor signal transducer and activator of transcription 3 (STAT3).
    [Show full text]
  • Vitamin D and Skeletal Muscle Strength and Endurance in COPD
    Eur Respir J 2013; 41: 309–316 DOI: 10.1183/09031936.00043112 CopyrightßERS 2013 Vitamin D and skeletal muscle strength and endurance in COPD Abigail S. Jackson*, Dinesh Shrikrishna*, Julia L. Kelly*, Nicholas Hart#, John Moxham", Michael I. Polkey*, Paul Kemp* and Nicholas S. Hopkinson* ABSTRACT: It is not known whether vitamin D levels make a significant contribution to muscle AFFILIATIONS dysfunction in chronic obstructive pulmonary disease (COPD). *Muscle Laboratory, National Institute for Health Research ¡SD ¡ In 104 COPD patients (mean forced expiratory volume in 1 s 44 22 % predicted) and 100 Respiratory Disease Biomedical age- and sex-matched controls, serum 25-hydroxyvitamin D (25(OH)D), 1,25-dihydroxyvitamin D Research Unit at the Royal Brompton (1,25(OH)2D) and parathyroid hormone (PTH) levels were measured and related to quadriceps and Harefield NHS Foundation Trust strength and endurance. In a subset of 26 patients and 13 controls, quadriceps biopsy was and Imperial College, London, #Guy’s and St Thomas’ NHS performed and mRNA expression of myogenic regulatory factors (mrf) and fibre-specific myosin Foundation Trust and King’s College heavy chains (MHC) was determined. London National Institute of Health COPD patients were weaker and less physically active than controls. 25(OH)D levels were Research Comprehensive Biomedical ¡ -1 ¡ -1 Research Centre, London, and similar in both groups (48.5 25.5 nmol?L COPD versus 55.4 28.3 nmol?L control, p50.07) but " ¡ -1 ¡ -1 Respiratory Muscle Laboratory, PTH levels were significantly higher in patients (5.2 2.3 pmol?mL versus 4.4 2.0 pmol?L , King’s College London School of p50.01).
    [Show full text]
  • Myogenic Regulatory Transcription Factors Regulate Growth In
    RESEARCH ARTICLE Myogenic regulatory transcription factors regulate growth in rhabdomyosarcoma Ineˆ s M Tenente1,2,3, Madeline N Hayes1,2, Myron S Ignatius1,2,4, Karin McCarthy1,2, Marielle Yohe5, Sivasish Sindiri5, Berkley Gryder5, Mariana L Oliveira1,2,6, Ashwin Ramakrishnan1,2, Qin Tang1,2, Eleanor Y Chen7, G Petur Nielsen8, Javed Khan5, David M Langenau1,2* 1Molecular Pathology, Cancer Center, and Regenerative Medicine, Massachusetts General Hospital, Boston, United States; 2Harvard Stem Cell Institute, Cambridge, United States; 3GABBA Program, Abel Salazar Biomedical Sciences Institute, University of Porto, Porto, Portugal; 4Molecular Medicine, Greehey Children’s Cancer Research Institute, San Antonio, United States; 5Oncogenomics Section, Pediatric Oncology Branch, Advanced Technology Center, National Cancer Institute, Gaithersburg, United States; 6Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal; 7Department of Pathology, University of Washington, Seattle, United States; 8Department of Pathology, Massachusetts General Hospital, Boston, United States Abstract Rhabdomyosarcoma (RMS) is a pediatric malignacy of muscle with myogenic regulatory transcription factors MYOD and MYF5 being expressed in this disease. Consensus in the field has been that expression of these factors likely reflects the target cell of transformation rather than being required for continued tumor growth. Here, we used a transgenic zebrafish model to show that Myf5 is sufficient to confer tumor-propagating potential to RMS cells and caused tumors to initiate earlier and have higher penetrance. Analysis of human RMS revealed that MYF5 and MYOD *For correspondence: are mutually-exclusively expressed and each is required for sustained tumor growth. ChIP-seq and [email protected] mechanistic studies in human RMS uncovered that MYF5 and MYOD bind common DNA regulatory elements to alter transcription of genes that regulate muscle development and cell cycle Competing interests: The progression.
    [Show full text]
  • Modulation of Osteogenic and Myogenic Differentiation By
    www.nature.com/scientificreports OPEN Modulation of osteogenic and myogenic diferentiation by a phytoestrogen formononetin via Received: 27 November 2018 Accepted: 12 June 2019 p38MAPK-dependent JAK-STAT Published: xx xx xxxx and Smad-1/5/8 signaling pathways in mouse myogenic progenitor cells Ilavenil Soundharrajan1, Da Hye Kim2, Palaniselvam Kuppusamy1 & Ki Choon Choi1 Formononetin (FN), a typical phytoestrogen has attracted substantial attention as a novel agent because of its diverse biological activities including, osteogenic diferentiation. However, the molecular mechanisms underlying osteogenic and myogenic diferentiation by FN in C2C12 progenitor cells remain unknown. Therefore the objective of the current study was to investigate the action of FN on myogenic and osteogenic diferentiation and its impact on signaling pathways in C2C12 cells. FN signifcantly increased myogenic markers such as Myogenin, myosin heavy chains, and myogenic diferentiation 1 (MyoD). In addition, the expression of osteogenic specifc genes alkaline phosphatase (ALP), Run-related transcription factor 2(RUNX2), and osteocalcin (OCN) were up-regulated by FN treatment. Moreover, FN enhanced the ALP level, calcium deposition and the expression of bone morphogenetic protein isoform (BMPs). Signal transduction pathways mediated by p38 mitogen- activated protein kinase (p38MAPK), extracellular signal-related kinases (ERKs), protein kinase B (Akt), Janus kinases (JAKs), and signal transducer activator of transcription proteins (STATs) in myogenic and osteogenic diferentiation after FN treatment were also examined. FN treatment activates myogenic diferentiation by increasing p38MAPK and decreasing JAK1-STAT1 phosphorylation levels, while osteogenic induction was enhanced by p38MAPK dependent Smad, 1/5/8 signaling pathways in C2C12 progenitor cells. Development of skeletal muscle cells is a strictly regulated process with diverse functions in organisms.
    [Show full text]
  • Deletion of the Sclerotome-Enriched Lncrna PEAT Augments Ribosomal Protein Expression
    Deletion of the sclerotome-enriched lncRNA PEAT augments ribosomal protein expression David A. Stafforda, Darwin S. Dichmanna, Jessica K. Changb, and Richard M. Harlanda,1 aDepartment of Molecular and Cell Biology, University of California, Berkeley, CA 94720; and bDepartment of Genetics, Stanford University, Stanford, CA 94305-5120 Contributed by Richard M. Harland, September 23, 2016 (sent for review July 22, 2016; reviewed by Margaret Buckingham and Chen-Ming Fan) To define a complete catalog of the genes that are activated during developing sclerotome by dissection from early embryos at 8- to mouse sclerotome formation, we sequenced RNA from embryonic 9.5-d postconception is technically difficult. Although a model mouse tissue directed to form sclerotome in culture. In addition to for presomitic mesoderm culture from stem cells has been de- well-known early markers of sclerotome, such as Pax1, Pax9,andthe veloped (10), this has not been exploited as an in vitro model of Bapx2/Nkx3-2 homolog Nkx3-1, the long-noncoding RNA PEAT (Pax1 the sclerotome. To obtain sufficient sclerotomal transcripts for enhancer antisense transcript) was induced in sclerotome-directed RNA-seq, we chemically treated explants of embryonic tissue to samples. Strikingly, PEAT is located just upstream of the Pax1 gene. bias differentiation of fluorescently marked presomitic mesoderm Using CRISPR/Cas9, we generated a mouse line bearing a complete toward the sclerotome. Following FACS, we compared control deletion of the PEAT-transcribed unit. RNA-seq on PEAT mutant and induced cultures to define the sclerotome transcriptome. In embryos showed that loss of PEAT modestly increases bone mor- addition to the previously described transcription factors, we identi- phogenetic protein target gene expression and also elevates the fied an uncharacterized noncoding RNA transcript that we named expression of a large subset of ribosomal protein mRNAs.
    [Show full text]
  • Supp Data.Pdf
    Supplementary Methods Verhaak sub-classification The Verhaak called tumor subtype was assigned as determined in the Verhaak et al. manuscript, for those TCGA tumors that were included at the time. TCGA data for expression (Affymetrix platform, level 1) and methylation (Infinium platform level 3) were downloaded from the TCGA portal on 09/29/2011. Affymetrix data were processed using R and Biocondutor with a RMA algorithm using quantile normalization and custom CDF. Level 3 methylation data has already been processed and converted to a beta-value. The genes that make up the signature for each of the 4 Verhaak groups (i.e. Proneural, Neural, Mesenchymal, and Classical) were downloaded. For each tumor, the averaged expression of the 4 genes signatures was calculated generating 4 ‘metagene’ scores, one for each subtype, (Colman et al.2010) for every tumor. Then each subtype metagene score was z-score corrected to allow comparison among the 4 metagenes. Finally, for each tumor the subtype with the highest metagene z-score among the four was used to assign subtype. Thus by this method, a tumor is assigned to the group to which it has the strongest expression signature. The shortcoming is that if a tumor has a ‘dual personality’ – for instance has strong expression of both classical and mesenchymal signature genes, there is some arbitrariness to the tumor’s assignment. Western Blot Analysis and cell fractionation Western blot analysis was performed using standard protocols. To determine protein expression we used the following antibodies: TAZ (Sigma and BD Biosciences), YAP (Santa Cruz Biotechnology), CD44 (Cell Signaling), FN1 (BD Biosciences), TEAD1, TEAD2, TEAD3, TEAD4 (Santa Cruz Biotechnology), MST1, p-MST1, MOB1, LATS1, LATS2, 14-3-3-, 1 ACTG2, (Cell Signaling), p-LATS1/2 (Abcam), Flag (Sigma Aldrich), Actin (Calbiochem), CAV2 (BD Biosciences), CTGF (Santa Cruz), RUNX2 (Sigma Aldrich), Cylin A, Cyclin E, Cyclin B1, p-cdk1, p-cdk4 (Cell Signaling Technologies).
    [Show full text]