Abstract
The blunt snout bream (Megalobrama amblycephala) is a typical hypoxia-sensitive fish, and hypoxia stress leads to reduced vitality and yield during aquaculture. To explore the specific adaptation mechanism under hypoxia, the blunt snout bream was treated with hypoxia (DO = 2.0 ± 0.1 mg/L) for 24 h, followed by 3 h of recovery. Our results depicted that the gill filament structure of blunt snout bream changed after hypoxia. During hypoxia for 24 h, the gill filament structure was altered, including a more than 80% expansion of the lamellar respiratory surface area and a proportionate apoptosis decrease in interlamellar cell mass (ILCM) volume. Thus, the water–blood diffusion distance was shortened to less than 46%. During hypoxia for 24 h, the activity of ROS in gill tissue increased significantly (p < 0.05), while the mitochondrial membrane potential decreased significantly (p < 0.05). During hypoxia, mRNA expression level of anti-apoptotic gene Bcl-2 in the gills of blunt snout bream decreased significantly (p < 0.05), while the expression of pro-apoptotic gene Bax mRNA increased significantly (p < 0.05). Thus, the ratio of Bax/Bcl-2 mRNA increased in the gills of blunt snout bream to promote the activity of Caspase-3. Together, our results indicated hypoxia-induced apoptosis in the gills of blunt snout bream through the mitochondrial pathway. In addition, a decreased expression of Phd1 and an increased expression of Hif-1α in gills under hypoxia stress indicates that blunt snout bream may cope with hypoxia-induced apoptosis by enhancing the HIF pathway. These results provide new insights into fish’s adaptation strategies and mechanisms of hypoxia.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.
Code availability
Not applicable.
References
Baldissera MD, Souza CD, Boaventura TP, Nakayama CL, Baldisserotto B, Luz RK (2020) Involvement of the phosphoryl transfer network in gill bioenergetic imbalance of pacama (Lophiosilurus alexandri) subjected to hypoxia: notable participation of creatine kinase. Fish Physiol Biochem 46(1):405–416. https://doi.org/10.1007/s10695-019-00728-0
Bratton SB, Walker G, Srinivasula SM, Sun XM, Butterworth M, Alnemri ES, Cohen GM (2001) Recruitment, activation and retention of caspases-9 and-3 by Apaf-1 apoptosome and associated XIAP complexes. Embo J 20(5):998–1009. https://doi.org/10.1093/emboj/20.5.998
Brauner CJ, Matey V, Zhang W, Richards JG, Dhillon RS, Cao ZD, Wang Y, Fu SJ (2011) Gill remodeling in crucian carp during sustained exercise and the effect on subsequent swimming performance. Physiol Biochem Zool 84(6):535–542. https://doi.org/10.1086/662664
Chen FY, Fu SY, Ma M, Chang L, Li XY (2023) Effects of hypoxia stress on the activity of antioxidant enzymes, neuronal apoptosis and expression of related genes of telencephalon in Gymnocypris przewalskii. Acta Hydrobiologica Sinica 47(4):572–580. https://doi.org/10.7541/2023.2021.085
Cregan SP, Fortin A, MacLaurin JG, Callaghan SM, Cecconi F, Yu SW, Slack RS (2002) Apoptosis-inducing factor is involved in the regulation of caspase-independent neuronal cell death. J Cell Biol 158(3):507–517. https://doi.org/10.1083/jcb.200202130
Danial NN, Korsmeyer SJ (2004) Cell death: critical control points. Cell 116(2):205–219. https://doi.org/10.1016/S0092-8674(04)00046-7
Dhillon RS, Yao L, Matey V, Chen BJ, Zhang AJ, Cao ZD, Richards JG (2013) Interspecific differences in hypoxia-induced gill remodeling in carp. Physiol Biochem Zool 86(6):727–739. https://doi.org/10.1086/673180
Eissing T, Waldherr S, Allgöwer F, Scheurich P, Bullinger E (2007) Response to bistability in apoptosis: roles of bax, bcl-2, and mitochondrial permeability transition pores. Biophys J 92(9):3332–3334. https://doi.org/10.1529/biophysj.106.100362
Erler JT, Cawthorne CJ, Williams KJ, Koritzinsky M, Wouters BG, Wilson C, Dive C (2004) Hypoxia-mediated down-regulation of Bid and Bax in tumors occurs via hypoxia-inducible factor 1-dependent and -independent mechanisms and contributes to drug resistance. Mol Cell Biol 24(7):2875–2889. https://doi.org/10.1128/MCB.24.7.2875-2889.2004
Esposito F, Tornincasa M, Chieffi P, DeMartino I, Pierantoni GM, Fusco A (2018) Retraction: high-mobility group a1 proteins regulate p53-mediated transcription of Bcl-2 gene. Cancer Res 78(24):6905. https://doi.org/10.1158/0008-5472.CAN-18-3454
Fariss MW, Chan CB, Patel M, Van HB, Orrenius S (2005) Role of mitochondria in toxic oxidative stress. Mol Interv 5(2):94–111. https://doi.org/10.1124/mi.5.2.7
Guan WZ, Guo DD, Sun YW, Chen J, Jiang XY, Zou SM (2017) Characterization of duplicated heme oxygenase-1 genes and their responses to hypoxic stress in blunt snout bream (Megalobrama amblycephala). Fish Physiol Biochem 43:641–651. https://doi.org/10.1007/s10695-016-0318-z
Hockenbery DM, Oltvai ZN, Yin XM, Milliman CL, Korsmeyer SJ (1993) Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75(2):241–251. https://doi.org/10.1016/0092-8674(93)80066-n
Hughes GM, Morgan M (1973) The structure of fish gills in relation to their respiratory function. Biol Rev 48(3):419–475. https://doi.org/10.1111/j.1469-185X.1973.tb01009.x
Ivan M, Kondo K, Yang HF, Kim W, Valiando J, Ohh M, Kaelin WG (2001) HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 292(5516):464–468. https://doi.org/10.1126/science.1059817
Klein JA, Ackerman SL (2003) Oxidative stress, cell cycle, and neurodegeneration. J Clin Invest 111(6):785–793. https://doi.org/10.1172/JCI18182
Lai Z, Zhang L, Su J, Cai D, Xu Q (2016) Sevoflurane postconditioning improves long-term learning and memory of neonatal hypoxia-ischemia brain damage rats via the PI3K/Akt-mPTP pathway. Brain Res 1630:25–37. https://doi.org/10.1016/j.brainres.2015.10.050
Law SH, Wu RS, Ng PK, Yu RM, Kong RY (2006) Cloning and expression analysis of two distinct HIF-alpha isoforms-gcHIF-1alpha and gcHIF-4alpha-from the hypoxia-tolerant grass carp, ctenopharyngodon idellus. Bmc Mol Biol 7(1):15. https://doi.org/10.1186/1471-2199-7-15
Li SF, Cai WQ, Zhou BY (1993) Variation in morphology and biochemical genetic markers among populations of blunt snout bream (Megalobrama amblycephala). Aquaculture 111(1–4):117–127. https://doi.org/10.1016/0044-8486(93)90030-3
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25(4):402–408. https://doi.org/10.1006/meth.2001.1262
Lu G, Mak YT, Wai SM, Kwong WH, Fang MR, James A, Yew DT (2005) Hypoxia-induced differential apoptosis in the central nervous system of the sturgeon (Acipenser shrenckii). Microsc Res Tech 68(5):258–263. https://doi.org/10.1002/jemt.20243
Matey V, Richards JG, Wang Y, Wood CM, Rogers J, Davies R, Murray BW, Chen XQ, Du J, Brauner CJ (2008) The effect of hypoxia on gill morphology and ionoregulatory status in the Lake Qinghai scaleless carp. Gymnocypris Przewalskii J Exp Biol 211(7):1063–1074. https://doi.org/10.1242/jeb.010181
Mcgettrick AF, O’Neill LAJ (2020) The role of HIF in immunity and inflammation. Cell Metab 32(4):524–536. https://doi.org/10.1016/j.cmet.2020.08.002
Nilsson GE (2007) Gill remodeling in fish - a new fashion or an ancient secret? J Exp Biol 210(14):2403–2409. https://doi.org/10.1242/jeb.000281
Nilsson GE, Dymowska A, Stecyk JA (2012) New insights into the plasticity of gill structure. Resp Physiol Neurobi 184(3):214–222. https://doi.org/10.1016/j.resp.2012.07.012
Pei X, Chu M, Tang P, Zhang H, Zhang X, Zheng X, Li J, Mei J, Wang T, Yin S (2021) Effects of acute hypoxia and reoxygenation on oxygen sensors, respiratory metabolism, oxidative stress, and apoptosis in hybrid yellow catfish “Huangyou-1.” Fish Physiol Biochem 47(5):1429–1448. https://doi.org/10.1007/s10695-021-00989-8
Phuong LM, Huong DTT, Nyengaard JR, Bayley M (2017) Gill remodelling and growth rate of striped catfish Pangasianodon hypophthalmus under impacts of hypoxia and temperature. Comp Biochem Phys A 203:288–296. https://doi.org/10.1016/j.cbpa.2016.10.006
Poli A, Beraudi A, Villani L, Storto M, Battaglia G, Gerevini VDG, Nicoletti F (2003) Group II metabotropic glutamate receptors regulate the vulnerability to hypoxic brain damage. J Neurosci 23(14):6023–6029. https://doi.org/10.1523/jneurosci.23-14-06023.2003
Poon WL, Hung CY, Nakano K, Randall DJ (2007) An in vivo study of common carp (Cyprinus carpio L.) liver during prolonged hypoxia. Comp Biochem Phys D 2(4):295–302. https://doi.org/10.1016/j.cbd.2007.05.002
Savitskaya MA, Onishchenko GE (2015) Mechanisms of apoptosis. Biochemistry (mosc) 80(11):1393–1405. https://doi.org/10.1134/S0006297915110012
Scherz-Shouval R, Shvets E, Fass E, Shorer H, Gil L, Elazar Z (2007) Reactive oxygen species are essential for autophagy and specifically regulate the activity of Atg4. Embo J 26(7):1749–1760. https://doi.org/10.1038/sj.emboj.7601623
Shuang L, Su XL, Zheng GD, Zou SM (2022) Effects of hypoxia and reoxygenation on gill remodeling, apoptosis, and oxidative stress in hypoxia-tolerant new variety blunt snout bream (Megalobrama amblycephala). Fish Physiol Biochem 48(1):263–274. https://doi.org/10.1007/s10695-022-01047-7
Sollid J, Angelis PD, Gundersen K, Nilsson GE (2003) Hypoxia induces adaptive and reversible gross morphological changes in crucian carp gills. J Exp Biol 206(20):3667–3673. https://doi.org/10.1242/jeb.00594
Springer JE, Nottingham SA, McEwen ML, Azbill RD, Jin Y (2001) Caspase-3 apoptotic signaling following injury to the central nervous system. Clin Chem Lab Med 39(4):299–307. https://doi.org/10.1515/CCLM.2001.046
Thomas LW, Ashcroft M (2019) Exploring the molecular interface between hypoxia-inducible factor signalling and mitochondria. Cell Mol Life Sci 76(9):1759–1777. https://doi.org/10.1007/s00018-019-03039-y
Tzaneva V, Vadeboncoeur C, Ting J, Perry SF (2014) Effects of hypoxia-induced gill remodelling on the innervation and distribution of ionocytes in the gill of goldfish, carassius auratus. J Comp Neurol 522(1):118–130. https://doi.org/10.1002/cne.23392
Wang M, Wu FL, Xie SG, Zhang L (2021) Acute hypoxia and reoxygenation: effect on oxidative stress and hypoxia signal transduction in the juvenile yellow catfish (Pelteobagrus fulvidraco). Aquaculture 531:735903. https://doi.org/10.1016/j.aquaculture.2020.735903
Williams TA, Bergstrome JC, Scott J, Bernier NJ (2017) CRF and urocortin 3 protect the heart from hypoxia/reoxygenation-induced apoptosis in zebrafish. Am J Physiol Regul Integr Comp Physiol 313(2):R91–R100. https://doi.org/10.1152/ajpregu.00045.2017
Wu CB, Zheng GD, Zhao XY, Zhou S, Zou SM (2020) Hypoxia tolerance in a selectively bred F4 population of blunt snout bream (Megalobrama amblycephala) under hypoxic stress. Aquaculture 518:734484. https://doi.org/10.1016/j.aquaculture.2019.734484
Youle RJ, Strasser A (2008) The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9(1):47–59. https://doi.org/10.1038/nrm2308
Yuan SJ, Akey CW (2013) Apoptosome structure, assembly, and procaspase activation. Structure 21(4):501–515. https://doi.org/10.1016/j.str.2013.02.024
Yuan ZH, Liu SK, Yao J, Zeng QF, Tan SX, Liu ZJ (2016) Expression of Bcl-2 genes in channel catfish after bacterial infection and hypoxia stress. Dev Comp Immunol 65:79–90. https://doi.org/10.1016/j.dci.2016.06.018
Zhao XY, Zhao SS, Zheng GD, Zhou JG, Zhou SM (2019) Functional conservation and divergence of duplicated the suppressor of cytokine signaling 1 in blunt snout bream (Megalobrama amblycephala). Gen Comp Endocr 284:113243. https://doi.org/10.1016/j.ygcen.2019.113243
Zhu CD, Wang ZH, Yan B (2013) Strategies for hypoxia adaptation in fish species: a review. J Comp Physiol B 183(8):1005–1013. https://doi.org/10.1007/s00360-013-0762-3
Funding
This work was supported by grants from the National Natural Science Foundation of China (32273100) and National Key Research and Development Plan of China.
Author information
Authors and Affiliations
Contributions
Xin-xin Yu, Yan-rui Zhang and Shan-shan Li experimented and related analyses. Xin-xin Yu wrote the manuscript. Shu-ming Zou and Guo-dong Zheng conceived the project and designed scientific objectives. All authors have read the paper and approved to submit it.
Corresponding author
Ethics declarations
Ethics Statement
All experiments were reviewed and carried out following the Guidelines for Humane Care and Treatment of Experimental Animals for Scientific Research, which drawn up by Ministry of Science and Technology, Beijing, China.
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing Interests
The authors declare that they have no conflict of interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Yu, Xx., Zhang, Yr., Li, Ss. et al. Effects of hypoxia on the gill morphological structure, apoptosis and hypoxia-related gene expression in blunt snout bream (Megalobrama amblycephala). Fish Physiol Biochem 49, 939–949 (2023). https://doi.org/10.1007/s10695-023-01233-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-023-01233-1