A biochemical basis for induction of retina regeneration by antioxidants
Nancy Echeverri-Ruiz, Tracy Haynes, Joseph Landers, Justin Woods, Michael J Gemma, Michael Hughes, Katia Del Rio-Tsonis, Nancy Echeverri-Ruiz, Tracy Haynes, Joseph Landers, Justin Woods, Michael J Gemma, Michael Hughes, Katia Del Rio-Tsonis
Abstract
The use of antioxidants in tissue regeneration has been studied, but their mechanism of action is not well understood. Here, we analyze the role of the antioxidant N-acetylcysteine (NAC) in retina regeneration. Embryonic chicks are able to regenerate their retina after its complete removal from retinal stem/progenitor cells present in the ciliary margin (CM) of the eye only if a source of exogenous factors, such as FGF2, is present. This study shows that NAC modifies the redox status of the CM, initiates self-renewal of the stem/progenitor cells, and induces regeneration in the absence of FGF2. NAC works as an antioxidant by scavenging free radicals either independently or through the synthesis of glutathione (GSH), and/or by reducing oxidized proteins through a thiol disulfide exchange activity. We dissected the mechanism used by NAC to induce regeneration through the use of inhibitors of GSH synthesis and the use of other antioxidants with different biochemical structures and modes of action, and found that NAC induces regeneration through its thiol disulfide exchange activity. Thus, our results provide, for the first time, a biochemical basis for induction of retina regeneration. Furthermore, NAC induction was independent of FGF receptor signaling, but dependent on the MAPK (pErk1/2) pathway.
Keywords: NAC; Regeneration; Retina.
Conflict of interest statement
Competing interests
No competing interests declared.
Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.
Figures
References
- Barford D. The role of cysteine residues as redox-sensitive regulatory switches. Curr Opin Struct Biol. 2004;14:679–686.
- Bely AE, Nyberg KG. Evolution of animal regeneration: re-emergence of a field. Trends Ecol Evol. 2010;25:161–170.
- Bloch K. The synthesis of glutathione in isolated liver. J Biol Chem. 1949;179:1245–1254.
- Cepko C. Intrinsically different retinal progenitor cells produce specific types of progeny. Nat Rev Neurosci. 2014;15:615–627.
- Corcoran A, Cotter TG. Redox regulation of protein kinases. FEBS J. 2013;280:1944–1965.
- Cotgreave IA. N-acetylcysteine: pharmacological considerations and experimental and clinical applications. Adv Pharmacol. 1997;38:205–227.
- Coulombre JL, Coulombre AJ. Regeneration of neural retina from the pigmented epithelium in the chick embryo. Dev Biol. 1965;12:79–92.
- Drowley L, Okada M, Beckman S, Vella J, Keller B, Tobita K, Huard J. Cellular antioxidant levels influence muscle stem cell therapy. Mol Ther. 2010;18:1865–1873.
- Edqvist PH, Myers SM, Hallbook F. Early identification of retinal subtypes in the developing, pre-laminated chick retina using the transcription factors Prox1, Lim1, Ap2alpha, Pax6, Isl1, Isl2, Lim3 and Chx10. Eur J Histochem. 2006;50:147–154.
- Fischer AJ, Bosse JL, El-Hodiri HM. The ciliary marginal zone (CMZ) in development and regeneration of the vertebrate eye. Exp Eye Res. 2013;116:199–204.
- Forman HJ, Fukuto JM, Torres M. Redox signaling: thiol chemistry defines which reactive oxygen and nitrogen species can act as second messengers. Am J Physiol-Cell Physiol. 2004;287:C246–C256.
- Galli S, Arciuch VGA, Poderoso C, Converso DP, Zhou QQ, Joffe EBD, Cadenas E, Boczkowski J, Carreras MC, Poderoso JJ. Tumor cell phenotype is sustained by selective MAPK oxidation in mitochondria. PLoS One. 2008;3
- Gupta V, Carroll KS. Sulfenic acid chemistry, detection and cellular lifetime. BBA-Gen Subj. 2014;1840:847–875.
- Hamburger V, Hamilton H. A series of normal stages in the development of the chick embryo. J Morphol. 1951;88:49–92.
- Haynes T, Gutierrez C, Aycinena JC, Tsonis PA, Del Rio-Tsonis K. BMP signaling mediates stem/progenitor cell-induced retina regeneration. Proc Natl Acad Sci USA. 2007;104:20380–20385.
- Haynes T, Luz-Madrigal A, Reis ES, Ruiz NPE, Grajales-Esquivel E, Tzekou A, Tsonis PA, Lambris JD, Del Rio-Tsonis K. Complement anaphylatoxin C3a is a potent inducer of embryonic chick retina regeneration. Nat Commun. 2013;4
- Ji AR, Ku SY, Cho MS, Kim YY, Kim YJ, Oh SK, Kim SH, Moon SY, Choi YM. Reactive oxygen species enhance differentiation of human embryonic stem cells into mesendodermal lineage. Exp Mol Med. 2010;42:175–186.
- Jopling C, Boue S, Izpisua Belmonte JC. Dedifferentiation, transdifferentiation and reprogramming: three routes to regeneration. Nat Rev Mol Cell Biol. 2011;12:79–89.
- Kang KW, Pak YM, Kim ND. Diethylmaleate and buthionine sulfoximine, glutathione-depleting agents, differentially inhibit expression of inducible nitric oxide synthase in endotoxemic mice. Nitric Oxide: Biol Chem. 1999;3:265–271.
- Kim KY, Rhim T, Choi I, Kim SS. N-acetylcysteine induces cell cycle arrest in hepatic stellate cells through its reducing activity. J Biol Chem. 2001;276:40591–40598.
- Kobayashi CI, Suda T. Regulation of reactive oxygen species in stem cells and cancer stem cells. J Cell Physiol. 2012;227:421–430.
- Koso H, Ouchi Y, Tabata Y, Aoki Y, Satoh S, Arai K, Watanabe S. SSEA-1 marks regionally restricted immature subpopulations of embryonic retinal progenitor cells that are regulated by the Wnt signaling pathway. Dev Biol. 2006;292:265–276.
- Lane SW, Williams DA, Watt FM. Modulating the stem cell niche for tissue regeneration. Nat Biotechnol. 2014;32:795–803.
- Laragione T, Bonetto V, Casoni F, Massignan T, Bianchi G, Gianazza E, Ghezzi P. Redox regulation of surface protein thiols: identification of integrin alpha-4 as a molecular target by using redox proteomics. Proc Natl Acad Sci USA. 2003;100:14737–14741.
- Leonard SE, Carroll KS. Chemical ‘omics’ approaches for understanding protein cysteine oxidation in biology. Curr Opin Chem Biol. 2011;15:88–102.
- Lonergan T, Bavister B, Brenner C. Mitochondria in stem cells. Mitochondrion. 2007;7:289–296.
- Luz-Madrigal A, Grajales-Esquivel E, McCorkle A, DiLorenzo AM, Barbosa-Sabanero K, Tsonis PA, Del Rio-Tsonis K. Reprogramming of the chick retinal pigmented epithelium after retinal injury. BMC Biol. 2014;12
- Murray TVA, Smyrnias I, Schnelle M, Mistry RK, Zhang M, Beretta M, Martin D, Anilkumar N, de Silva SM, Shah AM, Brewer AC. Redox regulation of cardiomyocyte cell cycling via an ERK1/2 and c-Myc-dependent activation of cyclin D2 transcription. J Mol Cell Cardiol. 2015;79:54–68.
- Niki E. Action of ascorbic-acid as a scavenger of active and stable oxygen radicals. Am J Clin Nutr. 1991;54:S1119–S1124.
- Ohori M, Kinoshita T, Yoshimura S, Warizaya M, Nakajima H, Miyake H. Role of a cysteine residue in the active site of ERK and the MAPKK family. Biochem Biophys Res Commun. 2007;353:633–637.
- Owusu-Ansah E, Yavari A, Mandal S, Banerjee U. Distinct mitochondrial retrograde signals control the G1-S cell cycle checkpoint. Nat Genet. 2008;40:356–361.
- Park C, Hollenberg M. Basic fibroblast growth factor induces retinal regeneration in vivo. Dev Biol. 1989;134:201–205.
- Rahman I, Kode A, Biswas SK. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat Protoc. 2006;1:3159–3165.
- Sart S, Song L, Li Y. Controlling redox status for stem cell survival, expansion, and differentiation. Oxid Med Cell Longev. 2015;2015:105135.
- Sehring IM, Jahn C, Weidinger G. Zebrafish fin and heart: what’s special about regeneration? Curr Opin Genet Dev. 2016;40:48–56.
- Serras F. The benefits of oxidative stress for tissue repair and regeneration. Fly. 2016;10:128–133.
- Spence JR, Aycinena JC, Del Rio-Tsonis K. Fibroblast growth factor-hedgehog interdependence during retina regeneration. Dev Dyn. 2007a;236:1161–1174.
- Spence JR, Aycinena JC, Del Rio-Tsonis K. Fibroblast growth factor-hedgehog interdependence during retina regeneration. Dev Dyn. 2007b;236:1161–1174.
- Spence JR, Madhavan M, Ewing JD, Jones DK, Lehman BM, Del Rio-Tsonis K. The hedgehog pathway is a modulator of retina regeneration. Development. 2004;131:4607–4621.
- Sun SY. N-acetylcysteine, reactive oxygen species and beyond. Cancer Biol Ther. 2010;9:109–110.
- Urao N, Ushio-Fukai M. Redox regulation of stem/progenitor cells and bone marrow niche. Free Radic Biol Med. 2013;54:26–39.
- Uzun MA, Koksal N, Kadioglu H, Gunerhan Y, Aktas S, Dursun N, Sehirli AO. Effects of N-acetylcysteine on regeneration following partial hepatectomy in rats with nonalcoholic fatty liver disease. Surg Today. 2009;39:592–597.
- Welin D, Novikova LN, Wiberg M, Kellerth JO, Novikov LN. Effects of N-acetyl-cysteine on the survival and regeneration of sural sensory neurons in adult rats. Brain Res. 2009;1287:58–66.
- Wipf P, Xiao JB, Jiang JF, Belikova NA, Tyurin VA, Fink MP, Kagan VE. Mitochondrial targeting of selective electron scavengers: synthesis and biological analysis of hemigramicidin-TEMPO conjugates. J Am Chem Soc. 2005;127:12460–12461.
- Xiao Y, Li X, Cui YQ, Zhang J, Liu LJ, Xie XY, Hao H, He GL, Kander MC, Chen MJ, Liu ZH, Verfaillie CM, Zhu H, Lei MX, Liu ZG. Hydrogen peroxide inhibits proliferation and endothelial differentiation of bone marrow stem cells partially via reactive oxygen species generation. Life Sci. 2014;112:33–40.
- Xiong LY, Sun JM, Hirche C, Yang J, Yang YQ, Xia Y, Lehnhardt M, Wang RR, Fu X. In vitro N-acetyl-L-cysteine promotes proliferation and suppresses interleukin-8 expression in adipose-derived stem cells. Aesthet Plast Surg. 2012;36:1260–1265.
- Yamada M, Tsukimura N, Ikeda T, Sugita Y, Att W, Kojima N, Kubo K, Ueno T, Sakurai K, Ogawa T. N-acetyl cysteine as an osteogenesis-enhancing molecule for bone regeneration. Biomaterials. 2013;34:6147–6156.
- Yan CYI, Greene LA. Prevention of PC12 cell death by N-acetylcysteine requires activation of the Ras pathway. J Neurosci. 1998;18:4042–4049.
- Young MR, Yang HS, Colburn NH. Promising molecular targets for cancer prevention: AP-1, NF-kappa B and Pdcd4. Trends Mol Med. 2003;9:36–41.
- Zafarullah M, Li WQ, Sylvester J, Ahmad M. Molecular mechanisms of N-acetylcysteine actions. Cell Mol Life Sci. 2003;60:6–20.
Source: PubMed