Embryonic stem (ES) cells differentiate in vitro into all cell lineages. We previously found that the p38 mitogen activated kinase (p38MAPK) pathway controls the commitment of ES cells toward either cardiomyogenesis (p38 on) or neurogenesis (p38 off ). In this study, we show that p38α knock-out ES cells do not differentiate into cardiac, endothelial, smooth muscle, and skeletal muscle lineages. Reexpression of p38MAPK in these cells partially rescues their mesodermal differentiation defects and corrects the high level of spontaneous neurogenesis of knock-out cells. Wild-type ES cells were treated with a p38MAPK-specific inhibitor during the differentiation process. These experiments allowed us to identify 2 early independent successive p38MAPK functions in the formation of mesodermal lineages. Further, the first one correlates with the regulation of the expression of Brachyury, an essential mesodermal-specific transcription factor, by p38MAPK. In conclusion, by genetic and biochemical approaches, we demonstrate that p38MAPK activity is essential for the commitment of ES cell into cardiac, endothelial, smooth muscle, and skeletal muscle mesodermal lineages.
Get full access to this article
View all access options for this article.
References
1.
KashyapV, RezendeN, ScotlandK, ShafferS, PerssonJ, GudasL, MonganN. 2009. Regulation of stem cell pluripotency and differentiation involves a mutual regulatory circuit of the NANOG, OCT4, and SOX2 pluripotency transcription factors with polycomb repressive complexes and stem cell microRNAs. Stem Cell Dev, 18:1093–1108.
CuendaA, RousseauS. 2007. p38 MAP-Kinases pathway regulation, function and role in human diseases. Biochim Biophys Acta—Mol Cell Res, 1773:1358–1375.
10.
MorookaT, NishidaE. 1998. Requirement of p38 mitogen-activated protein kinase for neuronal differentiation in PC12 cells. J Biol Chem, 273:24285–24288.
11.
RousseauS, HouleF, LandryJ, HuotJ. 1997. p38 MAP kinase activation by vascular endothelial growth factor mediates actin reorganization and cell migration in human endothelial cells. Oncogene, 15:2169–2177.
12.
CuendaA, CohenP. 1999. Stress-activated protein kinase-2/p38 and a rapamycin-sensitive pathway are required for C2C12 myogenesis. J Biol Chem, 274:4341–4346.
13.
TamuraK, SudoT, SenftlebenU, DadakAM, JohnsonR, KarinM. 2000. Requirement for p38alpha in erythropoietin expression: a role for stress kinases in erythropoiesis. Cell, 102:221–231.
14.
AdamsRH, PorrasA, AlonsoG, JonesM, VinterstenK, PanelliS, ValladaresA, PerezL, KleinR, NebredaAR. 2000. Essential role of p38alpha MAP kinase in placental but not embryonic cardiovascular development. Mol Cell, 6:109–116.
15.
AllenM, SvenssonL, RoachM, HamborJ, McNeishJ, GabelCA. 2000. Deficiency of the stress kinase p38alpha results in embryonic lethality: characterization of the kinase dependence of stress responses of enzyme-deficient embryonic stem cells. J Exp Med, 191:859–870.
16.
AouadiM, BostF, CaronL, LaurentK, Le Marchand BrustelY, BinetruyB. 2006. p38 Mitogen-activated protein kinase activity commits embryonic stem cells to either neurogenesis or cardiomyogenesis. Stem Cells, 24:1399–1406.
17.
BostF, CaronL, MarchettiI, DaniC, Le Marchand-BrustelY, BinetruyB. 2002. Retinoic acid activation of the ERK pathway is required for embryonic stem cell commitment into the adipocyte lineage. Biochem J, 361:621–627.
18.
KimJM, WhiteJM, ShawAS, SleckmanBP. 2005. MAPK p38{alpha} is dispensable for lymphocyte development and proliferation. J Immunol, 174:1239–1244.
19.
CaronL, BostF, ProtM, HofmanP, BinetruyB. 2005. A new role for the oncogenic high-mobility group A2 transcription factor in myogenesis of embryonic stem cells. Oncogene, 24:6281–6291.
20.
CaronL, ProtM, RouleauM, RolandoM, BostF, BinetruyB. 2005. The Lac repressor provides a reversible gene expression system in undifferentiated and differentiated embryonic stem cell. Cell Mol Life Sci, 62:1605–1612.
21.
DreuxM, DaoT, FresquetJ, GuerinM, JuliaZ, VerneyG, DurantelD, ZoulimF, LavilletteD, CossetF, BartoschB. 2009. Receptor complementation and mutagenesis reveal SR-BI as an essential HCV entry factor and functionally imply its intra- and extra-cellular domains. PLoS Pathog, 5:e1000310.
22.
NègreD, MangeotP, DuisitG, BlanchardS, VidalainP, LeissnerP, WinterA, Rabourdin-CombeC, MehtaliM, MoullierP, DarlixJ, CossetFL. 2000. Characterization of novel safe lentiviral vectors derived from simian immunodeficiency virus (SIVmac251) that efficiently transduce mature human dendritic cells. Gene Ther, 7:1613–1623.
23.
MurryCE, KellerG. 2008. Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development. Cell, 132:661–680.
24.
GimondC, MarchettiS, PagesG. 2006. Differentiation of mouse embryonic stem cells into endothelial cells: genetic selection and potential use in vivo. Methods Mol Biol, 330:303–329.
SoneM, ItohH, YamashitaJ, Yurugi-KobayashiT, SuzukiY, KondoY, NonoguchiA, SawadaN, YamaharaK, MiyashitaK, ParkK, ShibuyaM, NitoS, NishikawaS-I, NakaoK. 2003. Different differentiation kinetics of vascular progenitor cells in primate and mouse embryonic stem cells. Circulation, 107:2085–2088.
29.
AlexopoulouA, CouchmanJ, WhitefordJ. 2008. The CMV early enhancer/chicken beta actin (CAG) promoter can be used to drive transgene expression during the differentiation of murine embryonic stem cells into vascular progenitors. BMC Cell Biol, 9:2.
30.
ShowellC, BinderO, ConlonF. 2004. T-box genes in early embryogenesis. Dev Dyn, 229:201–218.
31.
FehlingHJr, LacaudG, KuboA, KennedyM, RobertsonS, KellerG, KouskoffV. 2003. Tracking mesoderm induction and its specification to the hemangioblast during embryonic stem cell differentiation. Development, 130:4217–4227.
32.
DuvalD, MalaiseM, ReinhardtB, KedingerC, BoeufH. 2004. A p38 inhibitor allows to dissociate differentiation and apoptotic processes triggered upon LIF withdrawal in mouse embryonic stem cells. Cell Death Differ, 11:331–341.
33.
ZhengM, ReynoldsC, JoS-H, WerstoR, HanQ, XiaoR-P. 2004. Intracellular acidosis-activated p38 MAPK signaling and its essential role in cardiomyocyte hypoxic injury. 10.1096/fj.04-2607fjeFASEB J04-2607fje.
34.
DavidsonSM, MorangeM. 2000. Hsp25 and the p38 MAPK pathway are involved in differentiation of cardiomyocytes. Dev Biol, 218:146–160.
35.
ErikssonM, LeppaS. 2002. Mitogen-activated protein kinases and activator protein 1 are required for proliferation and cardiomyocyte differentiation of p19 embryonal carcinoma cells. 10.1074/jbc.M107340200J Biol Chem, 277:15992–16001.
36.
GraichenR, XuX, BraamSR, BalakrishnanT, NorfizaS, SiehS, SooSY, ThamSC, MummeryC, ColmanA, ZweigerdtR, DavidsonBP. 2008. Enhanced cardiomyogenesis of human embryonic stem cells by a small molecular inhibitor of p38 MAPK. Differentiation, 76:357–370.
37.
Schmidt-LuckeC, RossigL, FichtlschererS, VasaM, BrittenM, KamperU, DimmelerS, ZeiherAM. 2005. Reduced number of circulating endothelial progenitor cells predicts future cardiovascular events: proof of concept for the clinical importance of endogenous vascular repair. Circulation, 111:2981–2987.
KellerG. 2005. Embryonic stem cell differentiation: emergence of a new era in biology and medicine. Genes Dev, 19:1129–1155.
40.
GarryDJ, OlsonEN. 2006. A common progenitor at the heart of development. Cell, 127:1101–1104.
41.
LluisF, PerdigueroE, NebredaAR, Munoz-CanovesP. 2006. Regulation of skeletal muscle gene expression by p38 MAP kinases. Trends Cell Biol, 16:36–44.
42.
de AngelisL, ZhaoJ, AndreucciJJ, OlsonEN, CossuG, McDermottJC. 2005. Regulation of vertebrate myotome development by the p38 MAP kinase-MEF2 signaling pathway. Dev Biol, 283:171–179.
43.
JonesNC, TynerKJ, NibargerL, StanleyHM, CornelisonDDW, FedorovYV, OlwinBB. 2005. The p38{alpha}/{beta} MAPK functions as a molecular switch to activate the quiescent satellite cell. 10.1083/jcb.200408066J Cell Biol, 169:105–116.
44.
JonesNC, FedorovYV, RosenthalRS, OlwinBB. 2001. ERK1/2 is required for myoblast proliferation but is dispensable for muscle gene expression and cell fusion. J Cell Physiol, 186:104–115.
45.
WuZ, WoodringPJ, BhaktaKS, TamuraK, WenF, FeramiscoJR, KarinM, WangJYJ, PuriPL. 2000. p38 and extracellular signal-regulated kinases regulate the myogenic program at multiple steps. Mol Cell Biol, 20:3951–3964.
46.
PerdigueroE, Ruiz-BonillaV, GreshL, HuiL, BallestarE, Sousa-VictorP, Baeza-RajaB, JardiM, Bosch-ComasA, EstellerM, CaellesC, SerranoAL, WagnerEF, Munoz-CanovesP. 2007. Genetic analysis of p38 MAP kinases in myogenesis: fundamental role of p38[alpha] in abrogating myoblast proliferation. EMBO J, 26:1245–1256.
47.
KunathT, Saba-El-LeilMK, AlmousailleakhM, WrayJ, MelocheS, SmithA. 2007. FGF stimulation of the Erk1/2 signalling cascade triggers transition of pluripotent embryonic stem cells from self-renewal to lineage commitment. Development, 134:2895–2902.
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
