Abstract
Oxidative stress is one of most common environmental stresses encountered by fish, especially during their fragile larval stage. More and more studies are aimed at understanding the antioxidant defense mechanism of fish larvae. Herein we characterized the early resistance of zebrafish larvae to oxidative stress and investigated the underlying transcriptional regulations using RNA-seq. We found that pre-exposure of zebrafish larvae to 2 mM H2O2 for 1 or 3 h significantly improved their survival under higher doses of H2O2 (3 mM), suggesting the antioxidant defenses of zebrafish larvae were rapidly built under pre-exposure of H2O2. Comparative transcriptome analysis showed that 310 (185 up and 125 down) and 512 (331 up and 181 down) differentially expressed genes were generated after 1 and 3 h of pre-exposure, respectively. KEGG enrichment analysis revealed that protein processing in endoplasmic reticulum is a highly enriched pathway; multiple genes (e.g., hsp70.1, hsp70.2, and hsp90aa1.2) encoding heat shock proteins in this pathway were sharply upregulated presumably to correct protein misfolding and maintaining the cellular normal functions during oxidative stress. More importantly, the Keap1/Nrf2 system-mediated detoxification enzyme system was significantly activated, which regulates the upregulation of target genes (e.g., gstp1, gsr, and prdx1) to scavenger reactive oxygen species, thereby defending against apoptosis. In addition, the MAPK, as a transmitter of stress signals, was activated, which may play an important role in activating antioxidant system in the early stages of oxidative stress. Altogether, these findings demonstrate that zebrafish larvae rapidly establish resistance to oxidative stress, and this involves changes in protein processing, stress signal transmission, and the activation of detoxification pathways.
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References
Amaral JD, Xavier JM, Steer CJ, Rodrigues CM (2010) Targeting the p53 pathway of apoptosis. Curr Pharm Des 16:2493–2503. https://doi.org/10.2174/138161210791959818
Bae H, You S, Lim W, Song G (2021) Flufenoxuron disturbs early pregnancy in pigs via induction of cell death with ER-mitochondrial dysfunction. J Hazard Mater 401:122996. https://doi.org/10.1016/j.jhazmat.2020.122996
Bianchi A, Moulin D, Hupont S, Koufany M, Netter P, Reboul P, Jouzeau JY (2014) Oxidative stress-induced expression of HSP70 contributes to the inhibitory effect of 15d-PGJ(2) on inducible prostaglandin pathway in chondrocytes. Free Radic Biol Med 76:114–126. https://doi.org/10.1016/j.freeradbiomed.2014.07.028
Biller JD, Takahashi LS (2018) Oxidative stress and fish immune system: phagocytosis and leukocyte respiratory burst activity. An Acad Bras Cienc 90:3403–3414. https://doi.org/10.1590/0001-3765201820170730
Blewett TA, Wood CM (2015) Salinity-dependent nickel accumulation and oxidative stress responses in the euryhaline killifish (Fundulus heteroclitus). Arch Environ Contam Toxicol 68:382–394. https://doi.org/10.1007/s00244-014-0115-6
Calabrese V, Cornelius C, Mancuso C, Pennisi G, Calafato S, Bellia F et al (2008) Cellular stress response: a novel target for chemoprevention and nutritional neuroprotection in aging, neurodegenerative disorders and longevity. Neurochem Res 33:2444–2471. https://doi.org/10.1007/s11064-008-9775-9
Chen HY, Wang PP, Du ZK, Wang GW, Gao SX (2018) Oxidative stress, cell cycle arrest, DNA damage and apoptosis in adult zebrafish (Danio rerio) induced by tris (1,3-dichloro-2-propyl) phosphate. Aquat Toxicol 194:37–45. https://doi.org/10.1016/j.aquatox.2017.11.001
Demple B (1991) Regulation of bacterial oxidative stress genes. Annu Rev Genet 25:315–337. https://doi.org/10.1146/annurev.ge.25.120191.001531
Desaint S, Luriau S, Aude JC, Rousselet G, Toledano MB (2004) Mammalian antioxidant defenses are not inducible by H2O2. J Biol Chem 279:31157–31163. https://doi.org/10.1074/jbc.M401888200
Hotamisligil GS, Davis RJ (2016) Cell signaling and stress responses. Cold Spring Harb Perspect Biol 8:a006072. https://doi.org/10.1101/cshperspect.a006072
Itoh K, Mimura J, Yamamoto M (2010) Discovery of the negative regulator of Nrf2, Keap1: a historical overview. Antioxid Redox Signal 13:1665–1678. https://doi.org/10.1089/ars.2010.3222
Jarvis RM, Hughes SM, Ledgerwood EC (2012) Peroxiredoxin 1 functions as a signal peroxidase to receive, transduce, and transmit peroxide signals in mammalian cells. Free Radic Biol Med 53:1522–1530. https://doi.org/10.1016/j.freeradbiomed.2012.08.001
Karin M, Liu ZG, Zandi E (1997) AP-1 function and regulation. Curr Opin Cell Biol 9:240–246. https://doi.org/10.1016/s0955-0674(97)80068-3
Kim DH, Jung IH, Kim DH, Park SW (2019) Knockout of longevity gene Sirt1 in zebrafish leads to oxidative injury, chronic inflammation, and reduced life span. PLoS ONE 14:e0220581. https://doi.org/10.1371/journal.pone.0220581
Kobayashi M, Yamamoto M (2006) Nrf2-Keap1 regulation of cellular defense mechanisms against electrophiles and reactive oxygen species. Adv Enzyme Regul 46:113–140. https://doi.org/10.1016/j.advenzreg.2006.01.007
Kurashova NA, Madaeva IM, Kolesnikova LI (2019) Expression of heat shock proteins HSP70 under oxidative stress. Adv Gerontol 32:502–508. https://doi.org/10.1134/S2079057020010099
Lenczowski JM, Dominguez L, Eder AM, King LB, Zacharchuk CM, Ashwell JD (1997) Lack of a role for Jun kinase and AP-1 in Fas-induced apoptosis. Mol Cell Biol 17:170–181. https://doi.org/10.1128/MCB.17.1.170
Lin SM, Li FJ, Yuangsoi B, Doolgindachbaporn S (2018) Effect of dietary phospholipid levels on growth, lipid metabolism, and antioxidative status of juvenile hybrid snakehead (Channa argus×Channa maculata). Fish Physiol Biochem 44:401–410. https://doi.org/10.1007/s10695-017-0443-3
Liu H, Lai W, Liu X, Yang H, Fang Y, Tian L et al (2021) Exposure to copper oxide nanoparticles triggers oxidative stress and endoplasmic reticulum (ER)-stress induced toxicology and apoptosis in male rat liver and BRL-3A cell. J Hazard Mater 401:123349. https://doi.org/10.1016/j.jhazmat.2020.123349
Long Y, Li L, Li Q, He X, Cui ZB (2012) Transcriptomic characterization of temperature stress responses in larval zebrafish. PLoS ONE 7:e37209. https://doi.org/10.1371/journal.pone.0037209
Long Y, Song GL, Yan JJ, He XZ, Li Q, Cui ZB (2013) Transcriptomic characterization of cold acclimation in larval zebrafish. BMC Genomics 14:612. https://doi.org/10.1186/1471-2164-14-612
Long Y, Yan JJ, Song GL, Li XH, Li XX, Li Q, Cui ZB (2015) Transcriptional events co-regulated by hypoxia and cold stresses in Zebrafish larvae. BMC Genomics 16:385. https://doi.org/10.1186/s12864-015-1560-y
Lushchak VI (2014) Free radicals, reactive oxygen species, oxidative stress and its classification. Chem Biol Interact 224:164–175. https://doi.org/10.1016/j.cbi.2014.10.016
Lushchak VI (2016) Contaminant-induced oxidative stress in fish: a mechanistic approach. Fish Physiol Biochem 42:711–747. https://doi.org/10.1007/s10695-015-0171-5
Madaeva IM, Kurashova NA, Semenova NV, Ukhinov EB, Kolesnikov SI, Kolesnikova LI (2020) Heat shock protein HSP70 in oxidative stress in apnea patients. Bull Exp Biol Med 169:695–697. https://doi.org/10.1007/s10517-020-04957-9
Matsuzawa A, Nishitoh H, Tobiume K, Takeda K, Ichijo H (2002) Physiological roles of ASK1-mediated signal transduction in oxidative stress- and endoplasmic reticulum stress-induced apoptosis: Advanced findings from ASK1 knockout mice. Antioxid Redox Signal 4:415–425. https://doi.org/10.1089/15230860260196218
Nakajima H, Nakajima-Takagi Y, Tsujita T, Akiyama SI, Wakasa T, Mukaigasa K et al (2011) Tissue-restricted expression of Nrf2 and its target genes in zebrafish with gene-specific variations in the induction profiles. PLoS ONE 6:e26884. https://doi.org/10.1371/journal.pone.0026884
Nitz LF, Pellegrin L, Maltez LC, Pinto D, Sampaio LA, Monserrat JM, Garcia L (2020) Temperature and hypoxia on oxidative stress responses in pacu Piaractus mesopotamicus. J Therm Biol 92:102682. https://doi.org/10.1016/j.jtherbio.2020.102682
Novoa B, Pereiro P, López-Muñoz A, Varela M, Forn-Cuní G, Anchelin M, Dios S, Romero A, Martinez-López A, Medina-Gali RM, Collado M, Coll J, Estepa A, Cayuela ML, Mulero V, Figueras A (2019) Rag1 immunodeficiency-induced early aging and senescence in zebrafish are dependent on chronic inflammation and oxidative stress. Aging Cell 18:e13020. https://doi.org/10.1111/acel.13020
Oakes KD, Hewitt LM, McMaster ME, Wood C, Munkittrick KR, Van Der Kraak GJ (2005) Oxidative stress and sex steroid levels in fish following short-term exposure to pulp-mill effluents. J Toxicol Environ Health A 68:267–286. https://doi.org/10.1080/15287390590895621
Ott M, Gogvadze V, Orrenius S, Zhivotovsky B (2007) Mitochondria, oxidative stress and cell death. Apoptosis 12:913–922. https://doi.org/10.1007/s10495-007-0756-2
Oyewole AO, Birch-Machin MA (2015) Mitochondria-targeted antioxidants. FASEB J 29:4766–4771. https://doi.org/10.1096/fj.15-275404
Park K, Han EJ, Ahn G, Kwak I (2020) Effects of thermal stress-induced lead (Pb) toxicity on apoptotic cell death, inflammatory response, oxidative defense, and DNA methylation in zebrafish (Danio rerio) embryos. Aquat Toxicol 224:105479. https://doi.org/10.1016/j.aquatox.2020.105479
Qi M, Elion EA (2005) MAP kinase pathways. J Cell Sci 118:3569–3572. https://doi.org/10.1242/jcs.02470
Rhee SG, Woo HA, Kil IS, Bae SH (2012) Peroxiredoxin functions as a peroxidase and a regulator and sensor of local peroxides. J Biol Chem 287:4403–4410. https://doi.org/10.1074/jbc.r111.283432
Roychowdhury P, Aftabuddin M, Pati MK (2021) Thermal stress-induced oxidative damages in the liver and associated death in fish, Labeo rohita. Fish Physiol Biochem 47:21–32. https://doi.org/10.1007/s10695-020-00880-y
Shi CM, Zhao H, Zhai XL, Chen YJ, Lin SM (2019) Linseed oil can decrease liver fat deposition and improve antioxidant ability of juvenile largemouth bass, Micropterus salmoides. Fish Physiol Biochem 45:1513–1521. https://doi.org/10.1007/s10695-019-00636-3
Storz G, Imlay JA (1999) Oxidative stress. Curr Opin Microbiol 2:188–194. https://doi.org/10.1016/s1369-5274(99)80033-2
Sun J, Nan GX (2016) The mitogen-activated protein kinase (MAPK) signaling pathway as a discovery target in stroke. J Mol Neurosci 59:90–98. https://doi.org/10.1007/s12031-016-0717-8
Suzuki T, Takagi Y, Osanai H, Li L, Takeuchi M, Katoh Y, Kobayashi M, Yamamoto M (2005) Pi class glutathione S-transferase genes are regulated by Nrf 2 through an evolutionarily conserved regulatory element in zebrafish. Biochem J 388:65–73. https://doi.org/10.1042/BJ20041860
Szyller J, Bil-Lula I (2021) Heat shock proteins in oxidative stress and ischemia/reperfusion injury and benefits from physical exercises: a review to the current knowledge. Oxid Med Cell Longev 2021:6678457. https://doi.org/10.1155/2021/6678457
Tang S, Ye S, Ma Y, Liang Y, Liang N, Xiao F (2020) Clusterin alleviates Cr(VI)-induced mitochondrial apoptosis in L02 hepatocytes via inhibition of Ca(2+)-ROS-Drp1-mitochondrial fission axis. Ecotoxicol Environ Saf 205:111326. https://doi.org/10.1016/j.ecoenv.2020.111326
Villeneuve NF, Lau A, Zhang DD (2010) Regulation of the Nrf2-Keap1 antioxidant response by the ubiquitin proteasome system: an insight into cullin-ring ubiquitin ligases. Antioxid Redox Signal 13:1699–1712. https://doi.org/10.1089/ars.2010.3211
Xu H, Liu E, Li Y, Li X, Ding C (2017) Transcriptome analysis reveals increases in visceral lipogenesis and storage and activation of the antigen processing and presentation pathway during the mouth-opening stage in zebrafish larvae. Int J Mol Sci 18:1634. https://doi.org/10.3390/ijms18081634
Xu H, Jiang Y, Li S, Xie L, Tao YX, Li Y (2020) Zebrafish oxr1a knockout reveals its role in regulating antioxidant defenses and aging. Genes (Basel) 11:1118. https://doi.org/10.3390/genes11101118
Xu H, Fan SQ, Guo W, Miao XM, Li Y (2021) Transcriptome analysis reveals the importance of exogenous nutrition in regulating antioxidant defenses during the mouth-opening stage in oviparous fsh. Fish Physiol Biochem 47:1087–1103. https://doi.org/10.1007/s10695-021-00954-5
Yang CW, Lim WS, Song GH (2020) Mediation of oxidative stress toxicity induced by pyrethroid pesticides in fish. Comp Biochem Physiol C Toxicol Pharmacol 234:108758. https://doi.org/10.1016/j.cbpc.2020.108758
Zang YW, Zheng ST, Tang F, Yang L, Wei XP, Kong D, Sun WX, Li W (2020) Heme oxygenase 1 plays a crucial role in swamp eel response to oxidative stress induced by cadmium exposure or Aeromonas hydrophila infection. Fish Physiol Biochem 46:1947–1963. https://doi.org/10.1007/s10695-020-00846-0
Zheng M, Wang X, Templeton LJ, Smulski DR, LaRossa RA, Storz G (2001) DNA microarray-mediated transcriptional profiling of the Escherichia coli response to hydrogen peroxide. J Bacteriol 183:4562–70. https://doi.org/10.1128/JB.183.15.4562-4570.2001
Zhou L, Jing YK, Styblo M, Chen Z, Waxman S (2005) Glutathione-S-transferase pi inhibits As2O3-induced apoptosis in lymphoma cells: involvement of hydrogen peroxide catabolism. Blood 105:1198–203. https://doi.org/10.1182/blood-2003-12-4299
Zhou YL, Guo JL, Tang RJ, Ma HJ, Chen YJ, Lin SM (2020) High dietary lipid level alters the growth, hepatic metabolism enzyme, and anti-oxidative capacity in juvenile largemouth bass Micropterus salmoides. Fish Physiol Biochem 46:125–134. https://doi.org/10.1007/s10695-019-00705-7
Zhu XS, Zhu L, Lang YP, Chen YS (2008) Oxidative stress and growth inhibition in the freshwater fish Carassius auratus induced by chronic exposure to sublethal fullerene aggregates. Environ Toxicol Chem 27:1979–85. https://doi.org/10.1897/07-573.1
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The research was supported by the Fundamental Research Funds for the Central Universities (NO. SWU-KQ22015).
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Conceptualization, H.X. and Y.L.; methodology, H.X. and X.-M.M.; software, H.X.; validation, H.X. and X.-M.M.; formal analysis, H.X., X.-M.M., and W.-B.W.; investigation, H.X., X.-M.M., and G.W.; resources, Y.L.; data curation, H.X. and X.-M.M.; writing—original draft preparation, H.X.; writing—review and editing, H.X. and Y.L.; visualization, H.X. and W.-B.W.; supervision, Y.L.; project administration, H.X.; funding acquisition, Y.L. All authors have read and agreed to the published version of the manuscript.
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Xu, H., Miao, XM., Wang, WB. et al. Transcriptome analysis reveals the early resistance of zebrafish larvae to oxidative stress. Fish Physiol Biochem 48, 1075–1089 (2022). https://doi.org/10.1007/s10695-022-01100-5
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DOI: https://doi.org/10.1007/s10695-022-01100-5