Abstract
During migration, anadromous fish may encounter the effects of temperature and salinity of the water. In this study, we examined the effects of cold stress (25, 21, 17, and 13 °C) and salinity stress (20, 10, and 0 ‰) on morphology, enzyme activity, and gene expression level in the gills sampled from river pufferfish (Takifugu fasciatus). This environmental bifactor stress experiment suggested that the activities of catalase, glutathione peroxidase and total superoxide dismutase, as well as the expression of the sod2, hsp90, and cirp genes, increased with decreasing temperature. Among them, cirp reached its peak expression level before individuals experienced the lowest temperature. The osmosis-related genes nka, ncc, nkcc and cftr, all showed an increasing trend with increasing salinity and decreasing temperature. When cold and salinity stress were enhanced, apoptosis-related genes bcl-2, caspase-3 and caspase-9 promoted apoptosis through downregulation, up-regulation, and up-regulation, respectively. The study of the gill morphology showed that an increase in salinity or a decrease in temperature mainly leads to serious vacuolar lesions in the gill filaments. The results also showed moderate salinity (10 ‰) may alleviate the gill stress responses caused by low temperature. This research exhibited the primary molecular responses of the anadromous fish T. fasciatus under cold and salinity challenges, which could serve as a valuable reference for studying the responses of migratory fish to environmental changes.
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The data sets during the current study are available from the corresponding author on reasonable request.
References
Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126. https://doi.org/10.1016/S0076-6879(84)05016-3
Arthur J (2001) The glutathione peroxidases. CMLS. Cell Mol Life Sci 57:1825–1835. https://doi.org/10.1007/PL00000664
Barrett RD, Paccard A, Healy TM, Bergek S, Schulte PM, Schluter D, Rogers SM (2011) Rapid evolution of cold tolerance in stickleback. Proc R Soc B 278(1703):233–238. https://doi.org/10.1098/rspb.2010.0923
Bergeron D, Beauseigle D, Bellemare G (1993) Sequence and expression of a gene encoding a protein with RNA-binding and glycine-rich domains in Brassica napus. Biochem Biophys Acta 1216(1):123–125. https://doi.org/10.1016/0167-4781(93)90047-h
Breves JP, Serizier SB, Goffin V, McCormick SD, Karlstrom RO (2013) Prolactin regulates transcription of the ion uptake Na+/Cl- cotransporter (ncc) gene in zebrafish gill. Mol Cell Endocrinol 369(1–2):98–106. https://doi.org/10.1016/j.mce.2013.01.021
Cappello T, Brandão F, Guilherme S, Santos MA, Maisano M, Mauceri A, Canário J, Pacheco M, Pereira P (2016) Insights into the mechanisms underlying mercury-induced oxidative stress in gills of wild fish (Liza aurata) combining 1H NMR metabolomics and conventional biochemical assays. Sci Total Environ 548–549:13–24. https://doi.org/10.1016/j.scitotenv.2016.01.008
Carpenter CD, Kreps JA, Simon AE (1994) Genes encoding glycine-rich Arabidopsis thaliana proteins with RNA-binding motifs are influenced by cold treatment and an endogenous circadian rhythm. Plant Physiol 104(3):1015–1025. https://doi.org/10.1104/pp.104.3.1015
Cengiz EI, Unlu E (2006) Sublethal effects of commercial deltamethrin on the structure of the gill, liver and gut tissues of mosquitofish, Gambusia affinis: A microscopic study. Environ Toxicol Pharmacol 21(3):246–253. https://doi.org/10.1016/j.etap.2005.08.005
Cheng CH, Ye CX, Guo ZX, Wang AL (2017) Immune and physiological responses of pufferfish (Takifugu obscurus) under cold stress. Fish Shellfish Immunol 64:137–145. https://doi.org/10.1016/j.fsi.2017.03.003
Chu P, Wang T, Sun YR, Chu MX, Wang HY, Zheng X, Yin SW (2021) Effect of cold stress on the mapk pathway and lipidomics on muscle of Takifugu fasciatus. Aquaculture 540(73669):1. https://doi.org/10.1016/j.aquaculture.2021.736691
De Leeuw F, Zhang T, Wauquier C, Huez G, Kruys V, Gueydan C (2007) The cold-inducible RNA-binding protein migrates from the nucleus to cytoplasmic stress granules by a methylation-dependent mechanism and acts as a translational repressor. Exp Cell Res 313(20):4130–4144. https://doi.org/10.1016/j.yexcr.2007.09.017
Evans DH, Piermarini PM, Choe KP (2005) The multifunctional fish gill: dominant site of gas exchange, osmoregulation, acid-base regulation, and excretion of nitrogenous waste. Physiol Rev 85(1):97–177. https://doi.org/10.1152/physrev.00050.2003
Feng CY, Rise ML (2010) Characterization and expression analyses of anti-apoptotic Bcl-2-like genes NR-13, Mcl-1, Bcl-X1, and Bcl-X2 in Atlantic cod (Gadus morhua). Mol Immunol 47(4):763–784. https://doi.org/10.1016/j.molimm.2009.10.011
Fu D, Hu Y, Chu P, Wang T, Chu M, Shi Y, Yin S, Zhu Y, Wang Y, Guo Z (2021) Histopathological and calreticulin changes in the liver and gill of Takifugu fasciatus demonstrate the effects of copper nanoparticle and copper sulphate exposure. Aquacul Rep 20:100662. https://doi.org/10.1016/j.aqrep.2021.100662
Gam L, Thanh Huong DT, Tuong DD, Phuong NT, Jensen FB, Wang T, Bayley M (2020) Effects of temperature on acid-base regulation, gill ventilation and air breathing in the clown knifefish, Chitala ornata. J Experim Biol. https://doi.org/10.1242/jeb.216481
Gibbons TC, McBryan TL, Schulte PM (2018) Interactive effects of salinity and temperature acclimation on gill morphology and gene expression in threespine stickleback. Comparative biochemistry and physiology. Part A Mol Integrat Physiol 221:55–62. https://doi.org/10.1016/j.cbpa.2018.03.013
Griffitt RJ, Weil R, Hyndman KA, Denslow ND, Powers K, Taylor D, Barber DS (2007) Exposure to copper nanoparticles causes gill injury and acute lethality in zebrafish (Danio rerio). Environ Sci Technol 41(23):8178–8186. https://doi.org/10.1021/es071235e
Hao L, Wang Z, Xing B (2009) Effect of sub-acute exposure to TiO2 nanoparticles on oxidative stress and histopathological changes in Juvenile Carp (Cyprinus carpio). J Environ Sci (china) 21(10):1459–1466. https://doi.org/10.1016/s1001-0742(08)62440-7
He J, Qiang J, Yang H, Xu P, Zhu ZX, Yang RQ (2015) Changes in the fatty acid composition and regulation of antioxidant enzymes and physiology of juvenile genetically improved farmed tilapia Oreochromis niloticus (L.), subjected to short-term low temperature stress. J Therm Biol 53:90–97. https://doi.org/10.1016/j.jtherbio.2015.08.010
Hefnawy AEG, Tórtora- Pérez JL (2009) The importance of selenium and the effects of its deficiency in animal health. Small Rumin Res 89(2):185–192. https://doi.org/10.1016/j.smallrumres.2009.12.042
Hiroi J, McCormick SD (2007) Variation in salinity tolerance, gill Na+/K+-ATPase, Na+/K+/2Cl- cotransporter and mitochondria-rich cell distribution in three salmonids Salvelinus namaycush, Salvelinus fontinalis and Salmo salar. J Exp Biol 210:1015–1024. https://doi.org/10.1242/jeb.002030
Hiroi J, McCormick SD, Ohtani-Kaneko R, Kaneko T (2005) Functional classification of mitochondrion-rich cells in euryhaline Mozambique tilapia (Oreochromis mossambicus) embryos, by means of triple immunofluorescence staining for Na+/K+-ATPase, Na+/K+/2Cl- cotransporter and CFTR anion channel. J Exp Biol 208:2023–2036. https://doi.org/10.1242/jeb.01611
Hui W, Liang G, Liu J, Yang H, Xu P (2015) Combined effects of temperature and salinity on yolk utilization in Nile tilapia (oreochromis niloticus). Aquacul Res 46:2418–2425. https://doi.org/10.1111/are.12401
Hwang TC, Kirk KL (2013) The CFTR Ion Channel: Gating, Regulation, and Anion Permeation. Cold Spring Harb Perspect Med 3(1):a009498. https://doi.org/10.1101/cshperspect.a009498
Hwang PP, Lee TH, Lin LY (2011) Ion regulation in fish gills: recent progress in the cellular and molecular mechanisms. Am J Physiol Regul Integrat Compar Physiol 301(1):R28–R47. https://doi.org/10.1152/ajpregu.00047.2011
Iwama GK, Thomas PT, Forsyth RB, Vijayan MM (1998) Heat shock protein expression in fish. Rev Fish Biol Fisheries 8(1):35–56. https://doi.org/10.1023/A:1008812500650
Johnston IA, Bennett AF (1996). Animals and Temperature: Experimental Investigations of Evolutionary Adaptation to Temperature. https://doi.org/10.1017/CBO9780511721854
Kaneko A, Zheng H, Nakano I, Yuan G, Fujimori H, Yoneyama K (1996) Long-term acoustic measurement of temperature variations in the Pacific North Equatorial Current. J Geophys Res 101(C7):16373–16380. https://doi.org/10.1029/96JC01053
Karnaky KJ (1986) Structure and Function of the Chloride Cell of Fundulus heteroclitus and Other Teleosts. Integrat Compar Biol 26(1):209–224. https://doi.org/10.1093/icb/26.1.209
Kato A, Doi H, Nakada T, Sakai H, Hirose S (2005) Takifugu obscurus is a euryhaline fugu species very close to Takifugu rubripes and suitable for studying osmoregulation. BMC Physiol 5:18. https://doi.org/10.1186/1472-6793-5-18
Kato A, Muro T, Kimura Y, Li S, Islam Z, Ogoshi M, Doi H, Hirose S (2011) Differential expression of Na+-Cl- cotransporter and Na+-K+-Cl- cotransporter 2 in the distal nephrons of euryhaline and seawater pufferfishes. Am J Physiol Regulat Integrat Compar Physiol 300(2): R284–R297. https://doi.org/10.1152/ajpregu.00725.2009
Kuida K (2000) Caspase-9. Int J Biochem Cell Biol 32(2):121–124. https://doi.org/10.1016/s1357-2725(99)00024-2
Kültz D, Li J, Gardel l, Sacchi AR, (2013) Quantitative molecular phenotyping of gill remodeling in a cichlid fish responding to salinity stress. Mol Cell Proteom MCP 12(12):3962–3975. https://doi.org/10.1074/mcp.M113.029827
Langenau DM, Jette C, Berghmans S, Palomero T, Kanki JP, Kutok JL, Look AT (2005) Suppression of apoptosis by bcl-2 overexpression in lymphoid cells of transgenic zebrafish. Blood 105(8):3278–3285. https://doi.org/10.1182/blood-2004-08-3073
Li W, Yang G, Li X, Sun T, Wang J (2019a) Cluster analysis of the relationship between carbon dioxide emissions and economic growth. J Clean Product 225:459–471. https://doi.org/10.1016/j.jclepro.2019a.03.220
Li X, Wang T, Yin S, Zhang G, Cao Q, Wen X, Zhang H, Wang D, Zhu W (2019b) The improved energy metabolism and blood oxygen-carrying capacity for pufferfish, Takifugu fasciatus, against acute hypoxia under the regulation of oxygen sensors. Fish Physiol Biochem 45(1):323–340. https://doi.org/10.1007/s10695-018-0565-2
Livak KJ, Schmittgen TD (2002) Analysis of relative gene expression data using real-time quantitative pcr. Methods 25(4):402–408. https://doi.org/10.1006/meth.2001.1262
Long Y, Yan J, Song G, Li X, Li X, Li Q, Cui Z (2015) Transcriptional events co-regulated by hypoxia and cold stresses in Zebrafish larvae. BMC Genom 16(1):385. https://doi.org/10.1186/s12864-015-1560-y
Lytle C, Xu JC, Biemesderfer D, Forbush B (1995) Distribution and diversity of Na-K-Cl cotransport proteins: a study with monoclonal antibodies. Am J Physiol 269(6 Pt 1):C1496–C1505. https://doi.org/10.1152/ajpcell.1995.269.6.C1496
Madeira D, Vinagre C, Costa PM, Diniz MS (2014) Histopathological alterations, physiological limits, and molecular changes of juvenile Sparus aurata in response to thermal stress. Mar Ecol Prog Ser 505:253–266. https://doi.org/10.3354/meps10794
Mansouri B, Maleki A, Davari B, Johari SA, Shahmoradi B, Mohammadi E, Shahsavari S (2016) Histopathological effects following short-term coexposure of Cyprinus carpio to nanoparticles of TiO2 and CuO. Environ Monit Assess 188(10):575. https://doi.org/10.1007/s10661-016-5579-6
Masroor W, Farcy E, Gros R, Lorin-Nebel C (2018) Effect of combined stress (salinity and temperature) in European sea bass Dicentrarchus labrax osmoregulatory processes. Comp Biochem Physiol Part A Mol Integr Physiol 215:45–54. https://doi.org/10.1016/j.cbpa.2017.10.019
McCord JM, Fridovich I (1969) Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biolog Chem 244(22):6049–6055. https://doi.org/10.1016/0019-2791(69)90289-4
McCormick SD (1993) Methods for nonlethal gill biopsy and measurement of Na+, K+-ATPase activity. Can J Fish Aquat Sci 50(3):656–658. https://doi.org/10.1139/f93-075
Niu H, Hu P, Cheng P, Chu X, Chen L (2017) The role of dusp1 downregulation in apoptosis of zebrafish zf4 cells under cold stress. J Fish Sci China 24(2017):995–1002. https://doi.org/10.3724/SP.J.1118.2017.16358
Parihar MS, Javeri T, Hemnani T, Dubey AK, Prakash P (1997) Responses of superoxide dismutase, glutathione peroxidase and reduced glutathione antioxidant defenses in gills of the freshwater catfish (Heteropneustes fossilis) to short-term elevated temperature. J Therm Biol 22(2):151–156. https://doi.org/10.1016/S0306-4565(97)00006-5
Payne JA, Iii BF (1995) Molecular characterization of the epithelial Na-K-Cl cotransporter isoforms. Curr Opin Cell Biol 7(4):493–503. https://doi.org/10.1016/0955-0674(95)80005-0
Porter AG, Jänicke RU (1999) Emerging roles of caspase-3 in apoptosis. Cell Death Differ 6(2):99–104. https://doi.org/10.1038/sj.cdd.4400476
Qiongqiong XU, Han B, Luo J, Yan LI, Hou Y, Peng HU (2016) Effects of cold stress on ros production and expression of mapk proteins in zebrafish zf4 cells. J Fish Sci China 23:771–776. https://doi.org/10.3724/SP.J.1118.2016.15488
Saoud IP, Kreydiyyeh S, Chalfoun A, Fakih M (2007) Influence of salinity on survival, growth, plasma osmolality and gill Na+-K+-ATPase activity in the rabbitfish Siganus rivulatus. J Exp Mar Biol Ecol 348(1):183–190. https://doi.org/10.1016/j.jembe.2007.05.005
Schaarschmidt T, Meyer E, Jürss K (1999) A comparison of transport-related gill enzyme activities and tissue-specific free amino acid concentrations of Baltic Sea (brackish water) and freshwater threespine sticklebacks, Gasterosteus aculeatus, after salinity and temperature acclimation. Mar Biol 135(4):689–697. https://doi.org/10.1007/s002270050670
Scott GR, Schulte PM (2005) Intraspecific variation in gene expression after seawater transfer in gills of the euryhaline killifish Fundulus heteroclitus. Compar Biochem Physiol Part A 141(2):176–182. https://doi.org/10.1016/j.cbpb.2005.05.002
Sedmak JJ, Grossberg SE (1977) A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem 79(1–2):544–552. https://doi.org/10.1016/0003-2697(77)90428-6
Shen H, Sun Y, Zhang J, Wang X (2005) Dynamic effects of low-level 2-nitro-4′-hydroxydiphenylamine on the induction of hsp70 in the fish gills. Res Environ Sci 018(004):87–90 https://doi.org/10.13198/j.res.2005.04.89.shenh.016
Shi Y, Zhang G, Zhu Y, Liu J (2010) Effects of photoperiod, temperature, and salinity on growth and survival of obscure puffer Takifugu obscurus larvae. Aquaculture 309(1):103–108. https://doi.org/10.1016/j.aquaculture.2010.09.004
Snoeckx LH, Cornelussen RN, Van Nieuwenhoven FA, Reneman RS, Van Der Vusse GJ (2001) Heat shock proteins and cardiovascular pathophysiology. Physiol Rev 81(4):1461–1497. https://doi.org/10.1152/physrev.2001.81.4.1461
Tse WK, Au DW, Wong CK (2006) Characterization of ion channel and transporter mRNA expressions in isolated gill chloride and pavement cells of seawater acclimating eels. Biochem Biophys Res Commun 346(4):1181–1190. https://doi.org/10.1016/j.bbrc.2006.06.028
Wang L, Wang XL, Ren Q, Zhang GS, Liang FF, Yin SW (2016a) Immune responses of two superoxide dismutases (SODs) after lipopolysaccharide or Aeromonas hydrophila challenge in pufferfish, Takifugu obscurus. Aquaculture. https://doi.org/10.1016/j.aquaculture.2016.03.016
Wang J, Zhu X, Huang X, Gu L, Chen Y, Yang Z (2016b) Combined effects of cadmium and salinity on juvenile Takifugu obscurus: cadmium moderates salinity tolerance; salinity decreases the toxicity of cadmium. Sci Rep 6:30968. https://doi.org/10.1038/srep30968
Wang PF, Zeng S, Xu P, Zhou L, Li GF (2016c) Two HSP90 genes in mandarin fish Siniperca chuatsi: identification, characterization and their specific expression profiles during embryogenesis and under stresses. Fish Physiol Biochem 42(4):1123–1136. https://doi.org/10.1007/s10695-016-0202-x
Wang J, Tang H, Zhang X, Xue X, Zhu X, Chen Y, Yang Z (2018) Mitigation of nitrite toxicity by increased salinity is associated with multiple physiological responses: A case study using an economically important model species, the juvenile obscure puffer (Takifugu obscurus). Environ Pollut 232:137–145. https://doi.org/10.1016/j.envpol.2017.09.026
Wang T, Wen X, Hu Y, Zhang X, Wang D, Yin S (2019) Copper nanoparticles induced oxidation stress, cell apoptosis and immune response in the liver of juvenile Takifugu fasciatus. Fish Shellfish Immunol 84:648–655. https://doi.org/10.1016/j.fsi.2018.10.053
Wen B, Jin SR, Chen ZZ, Gao JZ (2018) Physiological responses to cold stress in the gills of discus fish (Symphysodon aequifasciatus) revealed by conventional biochemical assays and GC-TOF-MS metabolomics. Sci Total Environ 640–641:1372–1381. https://doi.org/10.1016/j.scitotenv.2018.05.401
Wen X, Zhang X, Hu Y, Xu J, Wang T, Yin S (2019) iTRAQ-based quantitative proteomic analysis of Takifugu fasciatus liver in response to low-temperature stress. J Proteom 201:27–36. https://doi.org/10.1016/j.jprot.2019.04.004
Wen X, Chu P, Xu J, Wei X, Fu D, Wang T, Yin S (2021) Combined effects of low temperature and salinity on the immune response, antioxidant capacity and lipid metabolism in the pufferfish (Takifugu fasciatus). Aquaculture 531:735866. https://doi.org/10.1016/j.aquaculture.2020.735866
Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X (1997) Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 275(5303):1129–1132. https://doi.org/10.1126/science.275.5303.1129
Yang R, Dai Z, Chen S, Chen L (2011) MicroRNA-mediated gene regulation plays a minor role in the transcriptomic plasticity of cold-acclimated zebrafish brain tissue. BMC Genom 12:605. https://doi.org/10.1186/1471-2164-12-605
Yoo GY, Lee JY (2016) The effect of feeding frequency, water temperature, and stocking density on the growth of river puffer Takifugu obscurus reared in a zero-exchange water system. Fish Aquatic Sci 19:23. https://doi.org/10.1186/s41240-016-0024-x
Zhou C, Lyu LH, Miao HK, Bahr T, Zhang QY, Liang T, Zhou HB, Chen GR, Bai Y (2020) Redox regulation by SOD2 modulates colorectal cancer tumorigenesis through AMPK-mediated energy metabolism. Mol Carcinog 59(5):545–556. https://doi.org/10.1002/mc.23178
Zhu X, Bührer C, Wellmann S (2016) Cold-inducible proteins CIRP and RBM3, a unique couple with activities far beyond the cold. Cell Mol Life Sci CMLS 73(20):3839–3859. https://doi.org/10.1007/s00018-016-2253-7
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The authors thank all members of the laboratory for valuable discussions.
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This work was supported by National Natural Science Foundation of China [32172948, 31800436], the “JBGS” Project of Seed Industry Revitalization in Jiangsu Province [JBGS(2021)034], the National Key R & D Programme of China [2018YFD0900301], and the Natural Science Foundation (NSF) of Jiangsu Province of China [BK20210563, BK20180728].
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WT and YS co-conceived this study, and supervised the experiments; MS, LS, ZK and ZX performed the experiments; WX, LY, SY, BY and CJ conducted the data analysis and created figures and tables; MS, LS and ZX wrote the manuscript; WT, MS and LY has contributed to the revision and refinement of this article.
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All experiments were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals in China. The method of euthanasia followed the Chinese Laws for Animal Experimentation. This study was approved by the Ethics Committee of Experimental Animals at Nanjing Normal University (Grant No. SYXK 2015–0028, Jiangsu, China).
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Ma, S., Luo, S., Zhang, K. et al. Mitigation of low temperature stress by increased salinity is associated with multiple physiological responses in the gills of Takifugu fasciatus. Mar Biol 169, 141 (2022). https://doi.org/10.1007/s00227-022-04128-6
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DOI: https://doi.org/10.1007/s00227-022-04128-6