Mechanistic target of rapamycin complex 1 signaling regulates cell proliferation, cell survival, and differentiation in regenerating zebrafish fins

BMC Developmental Biology

Yutaka Kikuchi¹  Shunya Hozumi¹  Taishi Shiomi¹  Kentaro Hirose¹ 

[1] Department of Biological Science, Graduate School of Science, Hiroshima University, Kagamiyama 1-3-1, Higashi-Hiroshima 739-8526, Hiroshima, Japan

关键词: Differentiation;    Cell survival;    Cell proliferation;    Osteoblast;    Zebrafish;    Regeneration;    Fin;    Mechanistic target of rapamycin;   

Others  :  1084688
DOI  :  10.1186/s12861-014-0042-9

 received in 2014-07-10, accepted in 2014-11-25,  发布年份 2014

【 摘 要 】

Background

The mechanistic target of rapamycin complex1 (mTORC1) signaling pathway has been implicated in functions of multicellular processes, including cell growth and metabolism. Although recent reports showed that many signaling pathways, including Activin, Bmp, Fgf, sonic hedgehog, Insulin-like growth factor (IGF), Notch, retinoic acid, and Wnt, are implicated in non-mammalian vertebrate regeneration, also known as epimorphic regeneration, mTORC1 function remains unknown.

Results

To investigate the role of mTORC1 signaling pathway in zebrafish caudal fin, we examined the activation and function of mTORC1 signaling using an antibody against phosphorylated S6 kinase and a specific inhibitor, rapamycin. mTORC1 signaling is activated in proliferative cells of intra-ray and wound epidermal cells before blastema formation, as well as in proliferative blastema cells, wound epidermal cells, and osteoblasts during regenerative outgrowth. Before blastema formation, proliferation of intra-ray and wound epidermal cells is suppressed, but cell death is not affected by mTORC1 signaling inhibition with rapamycin. Moreover, rapamycin treatment inhibits blastema and wound epidermal cell proliferation and survival during blastema formation and regenerative outgrowth, as well as osteoblast proliferation and differentiation during regenerative outgrowth. We further determined that mTORC1 signaling is regulated through IGF-1 receptor/phosphatidylinositol-3 kinase and Wnt pathways during fin regeneration.

Conclusion

Taken together, our findings reveal that mTORC1 signaling regulates proliferation, survival, and differentiation of intra-ray cells, wound epidermis, blastema cells, and/or osteoblasts in various fin regeneration stages downstream of IGF and Wnt signaling pathways.

【 授权许可】

   
2014 Hirose et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Tanaka EM, Reddien PW: The cellular basis for animal regeneration. Dev Cell 2011, 21(1):172-185.
• [2]Gemberling M, Bailey TJ, Hyde DR, Poss KD: The zebrafish as a model for complex tissue regeneration. Trends Genet 2013, 29(11):611-620.
• [3]Nechiporuk A, Keating MT: A proliferation gradient between proximal and msxb-expressing distal blastema directs zebrafish fin regeneration. Development 2002, 129(11):2607-2617.
• [4]Akimenko MA, Marí-Beffa M, Becerra J, Géraudie J: Old questions, new tools, and some answers to the mystery of fin regeneration. Dev Dyn 2003, 226(2):190-201.
• [5]Poss KD, Keating MT, Nechiporuk A: Tales of regeneration in zebrafish. Dev Dyn 2003, 226(2):202-210.
• [6]Grotek B, Wehner D, Weidinger G: Notch signaling coordinates cellular proliferation with differentiation during zebrafish fin regeneration. Development 2013, 140(7):1412-1423.
• [7]Li J, Kim SG, Blenis J: Rapamycin: One Drug. Many Effects. Cell Metab 2014, 19(3):373-379.
• [8]Laplante M, Sabatini DM: mTOR signaling in growth control and disease. Cell 2012, 149(2):274-293.
• [9]Tal TL, Franzosa JA, Tanguay RL: Molecular signaling networks that choreograph epimorphic fin regeneration in zebrafish - a mini-review. Gerontology 2010, 56(2):231-240.
• [10]Wehner D, Cizelsky W, Vasudevaro MD, Ozhan G, Haase C, Kagermeier-Schenk B, Röder A, Dorsky RI, Moro E, Argenton F, Kühl M, Weidinger G: Wnt/β-Catenin Signaling Defines Organizing Centers that Orchestrate Growth and Differentiation of the Regenerating Zebrafish Caudal Fin. Cell Rep 2014, 6(3):467-481.
• [11]Stewart S, Gomez AW, Armstrong BE, Henner A, Stankunas K: Sequential and opposing activities of Wnt and BMP coordinate zebrafish bone regeneration. Cell Rep 2014, 6(3):482-498.
• [12]Gauron C, Rampon C, Bouzaffour M, Ipendey E, Teillon J, Volovitch M, Vriz S: Sustained production of ROS triggers compensatory proliferation and is required for regeneration to proceed. Sci Rep 2013, 3:2084.
• [13]Brown AM, Fisher S, Iovine MK: Osteoblast maturation occurs in overlapping proximal-distal compartments during fin regeneration in zebrafish. Dev Dyn 2009, 238(11):2922-2928.
• [14]Woods AL, Hall PA, Shepherd NA, Hanby AM, Waseem NH, Lane DP, Levison DA: The assessment of proliferating cell nuclear antigen (PCNA) immunostaining in primary gastrointestinal lymphomas and its relationship to histological grade, S + G2 + M phase fraction (flow cytometric analysis) and prognosis. Histopathology 1991, 19(1):21-27.
• [15]Johnson SL, Weston JA: Temperature-sensitive mutations that cause stage-specific defects in Zebrafish fin regeneration. Genetics 1995, 141(4):1583-1595.
• [16]Li N, Felber K, Elks P, Croucher P, Roehl HH: Tracking gene expression during zebrafish osteoblast differentiation. Dev Dyn 2009, 238(2):459-466.
• [17]Thoreen CC, Kang SA, Chang JW, Liu Q, Zhang J, Gao Y, Reichling LJ, Sim T, Sabatini DM, Gray NS: An ATP-competitive mammalian target of rapamycin inhibitor reveals rapamycin-resistant functions of mTORC1. J Biol Chem 2009, 284(12):8023-8032.
• [18]Chresta CM, Davies BR, Hickson I, Harding T, Cosulich S, Critchlow SE, Vincent JP, Ellston R, Jones D, Sini P, James D, Howard Z, Dudley P, Hughes G, Smith L, Maguire S, Hummersone M, Malagu K, Menear K, Jenkins R, Jacobsen M, Smith GC, Guichard S, Pass M: AZD8055 is a potent, selective, and orally bioavailable ATP-competitive mammalian target of rapamycin kinase inhibitor with in vitro and in vivo antitumor activity. Cancer Res 2010, 70(1):288-298.
• [19]Makky K, Tekiela J, Mayer AN: Target of rapamycin (TOR) signaling controls epithelial morphogenesis in the vertebrate intestine. Dev Biol 2007, 303(2):501-513.
• [20]Akimenko MA, Johnson SL, Westerfield M, Ekker M: Differential induction of four msx homeobox genes during fin development and regeneration in zebrafish. Development 1995, 121(2):347-357.
• [21]Urasaki A, Morvan G, Kawakami K: Functional dissection of the Tol2 transposable element identified the minimal cis-sequence and a highly repetitive sequence in the subterminal region essential for transposition. Genetics 2006, 174(2):639-649.
• [22]Thummel R, Burket CT, Hyde DR: Two different transgenes to study gene silencing and re-expression during zebrafish caudal fin and retinal regeneration. Sci World J 2006, 6(Suppl 1):65-81.
• [23]Hoptak-Solga AD, Nielsen S, Jain I, Thummel R, Hyde DR, Iovine MK: Connexin43 (GJA1) is required in the population of dividing cells during fin regeneration. Dev Biol 2008, 317(2):541-548.
• [24]Varga M, Sass M, Papp D, Takács-Vellai K, Kobolak J, Dinnyés A, Klionsky DJ, Vellai T: Autophagy is required for zebrafish caudal fin regeneration. Cell Death Differ 2014, 21(4):547-556.
• [25]Vlahos CJ, Matter WF, Hui KY, Brown RF: A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). J Biol Chem 1994, 269(7):5241-5248.
• [26]Mitsiades CS, Mitsiades NS, McMullan CJ, Poulaki V, Shringarpure R, Akiyama M, Hideshima T, Chauhan D, Joseph M, Libermann TA, García-Echeverría C, Pearson MA, Hofmann F, Anderson KC, Kung AL: Inhibition of the insulin-like growth factor receptor-1 tyrosine kinase activity as a therapeutic strategy for multiple myeloma, other hematologic malignancies, and solid tumors. Cancer Cell 2004, 5(3):221-230.
• [27]Chen B, Dodge ME, Tang W, Lu J, Ma Z, Fan CW, Wei S, Hao W, Kilgore J, Williams NS, Roth MG, Amatruda JF, Chen C, Lum L: Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer. Nat Chem Biol 2009, 5(2):100-107.
• [28]Mohammadi M, McMahon G, Sun L, Tang C, Hirth P, Yeh BK, Hubbard SR, Schlessinger J: Structures of the tyrosine kinase domain of fibroblast growth factor receptor in complex with inhibitors. Science 1997, 276(5314):955-960.
• [29]Favata MF, Horiuchi KY, Manos EJ, Daulerio AJ, Stradley DA, Feeser WS, Van Dyk DE, Pitts WJ, Earl RA, Hobbs F, Copeland RA, Magolda RL, Scherle PA, Trzaskos JM: Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J Biol Chem 1998, 273(29):18623-18632.
• [30]ten Freyhaus H, Huntgeburth M, Wingler K, Schnitker J, Bäumer AT, Vantler M, Bekhite MM, Wartenberg M, Sauer H, Rosenkranz S: Novel Nox inhibitor VAS2870 attenuates PDGF-dependent smooth muscle cell chemotaxis, but not proliferation. Cardiovasc Res 2006, 71(2):331-341.
• [31]Shimobayashi M, Hall MN: Making new contacts: the mTOR network in metabolism and signalling crosstalk. Nat Rev Mol Cell Biol 2014, 15(3):155-162.
• [32]Kujawski S, Lin W, Kitte F, Börmel M, Fuchs S, Arulmozhivarman G, Vogt S, Theil D, Zhang Y, Antos CL: Calcineurin regulates coordinated outgrowth of zebrafish regenerating fins. Dev Cell 2014, 28(5):573-587.
• [33]Chablais F, Jazwinska A: IGF signaling between blastema and wound epidermis is required for fin regeneration. Development 2010, 137(6):871-879.
• [34]Valvezan AJ, Huang J, Lengner CJ, Pack M, Klein PS: Oncogenic mutations in adenomatous polyposis coli (Apc) activate mechanistic target of rapamycin complex 1 (mTORC1) in mice and zebrafish. Dis Model Mech 2014, 7(1):63-71.
• [35]Inoki K, Ouyang H, Zhu T, Lindvall C, Wang Y, Zhang X, Yang Q, Bennett C, Harada Y, Stankunas K, Wang CY, He X, MacDougald OA, You M, Williams BO, Guan KL: TSC2 integrates Wnt and energy signals via a coordinated phosphorylation by AMPK and GSK3 to regulate cell growth. Cell 2006, 126(5):955-968.
• [36]Hirose K, Shimoda N, Kikuchi Y: Transient reduction of 5-methylcytosine and 5-hydroxymethylcytosine is associated with active DNA demethylation during regeneration of zebrafish fin. Epigenetics 2013, 8(9):899-906.
• [37]Hyde DR, Godwin AR, Thummel R: In vivo electroporation of morpholinos into the regenerating adult zebrafish tail fin.J Vis Exp 2012, Mar 29 (61):e3632.
• [38]Kim SH, Kowalski ML, Carson RP, Bridges LR, Ess KC: Heterozygous inactivation of tsc2 enhances tumorigenesis in p53 mutant zebrafish. Dis Model Mech 2013, 6(4):925-933.
• [39]Boglev Y, Badrock AP, Trotter AJ, Du Q, Richardson EJ, Parslow AC, Markmiller SJ, Hall NE, de Jong-Curtain TA, Ng AY, Verkade H, Ober EA, Field HA, Shin D, Shin CH, Hannan KM, Hannan RD, Pearson RB, Kim SH, Ess KC, Lieschke GJ, Stainier DY, Heath JK: Autophagy induction is a Tor- and Tp53-independent cell survival response in a zebrafish model of disrupted ribosome biogenesis. PLoS Genet 2013, 9(2):e1003279.
• [40]Westerfield M: The zebrafish book. University of Oregon Press, Eugene; 1995.