Abstract
Nitric oxide (NO) is an important signalling molecule that plays diverse physiological functions in several vertebrates including that of adaptation to various stressful stimuli. The air-breathing magur catfish (Clarias magur) is known to tolerate a very high external ammonia (HEA) stress in its natural habitats. We report here the possible induction of inducible nitric oxide (inos) gene and more generation of NO in magur catfish exposed to HEA. Exposure to HEA (25 mM NH4Cl) for 14 days led to the higher accumulation of NO in different tissues of magur catfish and also more efflux of NO from the perfused liver of NH4Cl-treated fish as a consequence of high build of toxic ammonia in body tissues. More synthesis and accumulation of NO in body tissues was associated with the induction of iNOS activity, which otherwise was not detectable in control fish. The stimulation of iNOS activity in HEA exposed fish was mainly due to induction of inos gene as evidenced by more expression of inos mRNA and also more abundance of iNOS protein in different tissues of magur catfish. Immunocytochemical analysis indicated the zonal specific expression of iNOS protein in different tissues of magur catfish. The augmentation of iNOS in the fish under HEA could be an adaptive strategy of the fish to defend against the ammonia stress through the generation of NO. Therefore, the present finding identifies the potential role of iNOS to enhance the adaptive capacity and survivability of catfish under various adverse environmental and pathological conditions that it faces in its natural habitats.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amelio D, Garofalo F, Pellegrino D, Giordano F, Tota B, Cerra MC (2006) Cardiac expression and distribution of nitric oxide synthases in the ventricle of the cold-adapted Antarctic teleosts, the hemoglobinless Chionodraco hamatus and the red-blooded Trematomus bernacchii. Nitric Oxide 15:190–198. https://doi.org/10.1016/j.niox.2005.12.007
Amelio D, Garofalo F, Brunelli E, Loong AM, Wong WP, Ip YW, Tota B, Cerra MC (2008) Differential NOS expression in freshwater and aestivating Protopterus dolli (lungfish): heart vs kidney readjustments. Nitric Oxide 18:1–10. https://doi.org/10.1016/j.niox.2007.10.004
Andreakis N, D’Aniello S, Albalat R, Patti FP, Garcia-Fernàndez J, Procaccini G, Sordino P, Palumbo A (2011) Evolution of the nitric oxide synthase family in metazoans. Mol Biol Evol 28(1):163–179. https://doi.org/10.1093/molbev/msq179
Banerjee B, Bhuyan G, Koner D, Saha N (2018) Differential expression of multiple glutamine synthetase genes in air-breathing magur catfish, Clarias magur and their induction under hyper-ammonia stress. Gene 671:85–95. https://doi.org/10.1016/j.gene.2018.05.111
Barroso JB, Carreras A, Esteban FJ, Peinado MA, Martinez-Lara E, Valderrama R, Jimenez A, Rodrigo J, Lupianez JA (2000) Molecular and kinetic characterization and cell type location of inducible nitric oxide synthase in fish. Am J Physiol Regul Integr Comp Physiol 279:R650–R656. https://doi.org/10.1152/ajpregu.2000.279.2.R650
Bogdan C (2001) Nitric oxide and the immune response. Nat Immunol 2(10):907–916. https://doi.org/10.1038/ni1001-907
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Brüne B, Knethen AV, Sandau KB (1998) Nitric oxide and its role in apoptosis. Eur J Pharmacol 351(3):261–272. https://doi.org/10.1016/s0014-2999(98)00274-x
Cheng CH, Yang FF, Ling RZ, Liao SA, Miao YT, Ye CX, Wang AL (2015) Effects of ammonia exposure on apoptosis, oxidative stress and immune response in pufferfish (Takifugu obscurus). Aquat Toxicol 164:61–71. https://doi.org/10.1016/j.aquatox.2015.04.004
Choudhury MG, Saha N (2012a) Influence of environmental ammonia on the production of nitric oxide and expression of inducible nitric oxide synthase in the freshwater air-breathing catfish (Heteropneustes fossilis). Aquat Toxicol 116-117:43–53. https://doi.org/10.1016/j.aquatox.2012.03.006
Choudhury MG, Saha N (2012b) Expression of inducible nitric oxide synthase and nitric oxide in the mud-dwelled air-breathing singhi catfish, Heteropneustes fossilis under condition of water shortage. Nitric Oxide 27:219–227. https://doi.org/10.1016/j.niox.2012.07.006
Choudhury MG, Kumar S, Saha N. (2018) Role of nuclear factor kappa B and mitogen activated protein kinases in the induction of inducible nitric oxide synthase and nitric oxide production in primary hepatocytes of air-breathing catfish, Clarias magur under hyper-ammonia stress. (Submitted)
Coscelli C, Bermudez R, Ronza P, Losada AP, Quiroga MI (2016) Immunohistochemical study of inducible nitric oxide synthase and tumour necrosis factor alpha response in turbot (Scophthalmus maximus) experimentally infected with Aeromonas salmonicida subsp. Salmonicida. Fish Shellfish Immunol 56:294–302. https://doi.org/10.1016/j.fsi.2016.07.022
Demple B (2002) Signal transduction by nitric oxide in cellular stress responses. Mol Cell Biochem 234-235(1–2):11–18
Dini L, Lanubile R, Tarantino P, Mandich A, Cataldi E (2006) Expression of stress proteins 70 in tilapia (Oreochromis mossambicus) during confinement and crowding stress. Italian J Zool 73:117–124. https://doi.org/10.1080/11250000600679652
Giraldi-Guimarães A, Batista CM, Carneiro K, Tenório F, Cavalcante LA, Mendez-Otero R (2007) A critical survey on nitric oxide synthase expression and nitric oxide function in the retinotectal system. Brain Res Rev 56(2):403–426. https://doi.org/10.1016/j.brainresrev.2007.09.005
Gong L, Pitari GM, Schulz S, Waldman SA (2004) Nitric oxide signaling: systems integration of oxygen balance in defense of cell integrity. Curr Opin Hematol 11(1):7–14
Griffith OW, Stuehr DJ (1995) Nitric oxide synthases: properties and catalytic mechanism. Annu Rev Physiol 57:707–736. https://doi.org/10.1146/annurev.ph.57.030195.003423
Hensen MN, Jensen FB (2010) Nitric oxide metabolites in goldfish under normoxic and hypoxic conditions. J Exp Biol 213:3593–3602. https://doi.org/10.1242/jeb.048140
Ip YK, Chew SF, Randal DJ (2001) Ammonia toxicity, tolerance, and excretion. In: Wright P, Anderson PM (eds) Fish physiology. Academic Press, New York, pp 109–148. https://doi.org/10.1016/S1546-5098(01)20005-3
Ito K, Chen J, Khodadadian JJ, Seshan SV, Eaton C, Zhao X, Vaughan ED, Lipkowitz M, Poppas DP, Felsen D (2004) Liposome-mediated transfer of nitric oxide synthase gene improves renal function in ureteral obstruction in rats. Kidney Int 66:1365–1375. https://doi.org/10.1111/j.1523-1755.2004.00899.x
Iwama GK, Afonso LOB, Vijayan MM (2006) Stress in fishes. In: Evans DH, Claiborne JB (eds) The physiology of fishes. CRC Press, Boca Raton, pp 319–343
Jaffrey SR, Snyder SH (1995) Nitric oxide: a neural messenger. Annu Rev Cell Dev Biol 11:417–440. https://doi.org/10.1146/annurev.cb.11.110195.002221
Joyner MJ, Dietz NM (1997) Nitric oxide and vasodilation in human limbs. J Appl Physiol 83(6):1785–1796. https://doi.org/10.1152/jappl.1997.83.6.1785
Knowles RG, Salter M (1998) Measurement of NOS activity by conversion of radiolabeled arginine to citrulline using ion-exchange separation. Methods Mol Biol 100:67–73
Kun E, Kearney EB (1974) Ammonia. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol IV. Academic Press, New York, pp 1802–1806
Li M, Gong S, Li Q, Yuan L, Meng F, Wang R (2016) Ammonia toxicity induces glutamine accumulation, oxidative stress and immunosuppression in juvenile yellow catfish Pelteobagrus fulvidraco. Comp Biochem Physiol C 183-184:1–6. https://doi.org/10.1016/j.cbpc.2016.01.005
Losada AP, Bermúdez R, Faílde LD, Quiroga M (2012) Quantitative and qualitative evaluation of iNOS expression in turbot (Psetta maxima) infected with Enteromyxum scophthalmi. Fish Shellfish Immunol 32(2):243–248. https://doi.org/10.1016/j.fsi.2011.11.007
Malyshev IY, Manukhina EB (1998) Stress, adaptation and nitric oxide. Biochem 63(7):840–853
Milsom WK, Reid SG, Meier JT, Kinkead R (1999) Central respiratory pattern generation in the bullfrog, Rana catesbeiana. Comp Biochem Physiol A 124(3):253–264. https://doi.org/10.1016/S1095-6433(99)00113-0
Napoli C, Paolisso G, Casamassimi A, Al-Omran M, Barbieri M, Sommese L, Infante T, Ignarro LJ (2013) Effects of nitric oxide on cell proliferation: novel insights. J Am Coll Cardiol 62(2):89–95. https://doi.org/10.1016/j.jacc.2013.03.070
Nathan C, Xie QW (1994) Regulation of biosynthesis of nitric oxide. J Biol Chem 269:13725–13728
Pellegrino D, Sprovieri E, Mazza R, Randall DJ, Tota B (2002) Nitric oxide-cGMP-mediated vasoconstriction and effects of acetylcholine in the branchial circulation of the eel. Comp Biochem Physiol A 132(2):447–457
Pellegrino D, Palmerini CA, Tota B (2004) No hemoglobin but NO: the icefish (Chionodraco hamatus) heart as a paradigm. J Exp Biol 207:3855–3864. https://doi.org/10.1242/jeb.01180
Pietsch C, Vogt R, Neumann N, Kloas W (2008) Production of nitric oxide by carp (Cyprinus carpio L.) kidney leukocytes is regulated by cyclic 3′,5′-adenosine monophosphate. Comp Biochem Physiol A 150(1):58–65. https://doi.org/10.1016/j.cbpa.2008.03.003
Rio DC, Ares M, Hannon GJ, Nilsen TW (2010) Purification of RNA using TRIzol (TRI reagent). Cold Spring Harb Protoc 5439. https://doi.org/10.1101/pdb.prot5439
Saeij JP, Stet RJ, Groeneveld A, Verburg-van Kemenade LB, van Muiswinkel WB, Wiegertjes GF (2000) Molecular and functional characterization of a fish inducible-type nitric oxide synthase. Immunogenetics 51:339–346
Saha N, Ratha BK (1989) Comparative studies of ureogenesis in freshwater, air-breathing teleosts. Comp Physiol Biochem A 252(1):1–8. https://doi.org/10.1002/jez.1402520102
Saha N, Ratha BK (1998) Ureogenesis in Indian air-breathing teleosts: adaptation to environmental constraints. Comp Biochem Physiol A 120(2):195–208. https://doi.org/10.1016/S1095-6433(98)00026-9
Saha N, Ratha BK (2007) Functional ureogenesis to ammonia metabolism in Indian freshwater air-breathing catfishes. Fish Physiol Biochem 33:283–295. https://doi.org/10.1007/s10695-007-9172-3
Saha N, Dkhar J, Ratha BK (1995) Induction of ureogenesis in perfused liver of a freshwater teleost, Heteropneustes fossilis, infused with different concentrations of ammonium chloride. Comp Biochem Physiol B 112:733–741. https://doi.org/10.1016/0305-0491(95)00115-8
Saha N, Dutta S, Bhattacharjee A (2002) Role of amino acid metabolism in an air-breathing catfish, Clarias batrachus in response to exposure to a high concentration of exogenous ammonia. Comp Biochem Physiol B 133(2):235–250. https://doi.org/10.1016/S1096-4959(02)00145-8
Saha N, Datta S, Biswas K, Kharbuli ZY (2003) Role of ureogenesis in tackling problems of ammonia toxicity during exposure to higher ambient ammonia in the air-breathing walking catfish Clarias batrachus. J Biosci 28:733–742
Saha N, Datta S, Kharbuli ZY, Biswas K, Bhattacharjee A (2007) Air-breathing catfish, Clarias batrachus upregulates glutamine synthetase and carbamyl phosphate synthetase III during exposure to high external ammonia. Comp Biochem Physiol B 147(3):520–530. https://doi.org/10.1016/j.cbpb.2007.03.007
Schliess F, Görg B, Fischer R, Desjardins P, Bidmon JG, Herrmann A, Butterworth FR, Zilles K, Häussinger D (2002) Ammonia induces MK-801-sensitive nitration and phosphorylation of protein of protein tyrosine residues in rat astrocytes. FASEB J 16:739–741. https://doi.org/10.1096/fj.01-0862fje
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3(6):1101–1108
Sessa WC, Pritchard K, Seyedi N, Wang J, Hintze TH (1994) Chronic exercise in dogs increases coronary vascular nitric oxide production and endothelial cell nitric oxide synthase gene expression. Circ Res 74(2):349–353. https://doi.org/10.1161/01.RES.74.2.349
Sinha KA, AbdElgawad H, Giblen T, Zinta G, Rop DM, Asard H, Blust R, Boeck DG (2014) Anti-oxidative Defence are modulated differentially in three freshwater teleost in response to ammonia induced oxidative stress. PLoS One 9(4):e95319. https://doi.org/10.1371/journal.pone.0095319
Sinke AP, Jayakumar AR, Panickar KS, Moriyama M, Reddy PV, Norenberg MD (2008) NFkB in the mechanism of ammonia-induced astrocyte swelling in culture. J Neurochem 106:2302–2311. https://doi.org/10.1111/j.1471-4159.2008.05549.x
Tatsumi T, Matoba S, Kawahara A, Keira N, Shiraishi J, Akashi K, Kobara M, Tanaka T, Katamura M, Nakagawa C, Ohta B, Shirayama T, Takeda K, Asayama J, Fliss H, Nakagawa M (2000) Cytokine-induced nitric oxide production inhibits mitochondrial energy production and impairs contractile function in rat cardiac. J Am Coll Cardiol 35(5):1338–1346. https://doi.org/10.1016/S0735-1097(00)00526-X
Tota B, Amelio D, Pellegrino D, Ip YK, Cerra MC (2005) NO modulation of myocardial performance in fish hearts. Comp Biochem Physiol A 142(2):164–177. https://doi.org/10.1016/j.cbpb.2005.04.019
Trischitta F, Pidalà P, Faggio C (2007) Nitric oxide modulates ionic transport in the isolated intestine of the eel, Anguilla anguilla. Comp Biochem Physiol A 148(2):368–373. https://doi.org/10.1016/j.cbpa.2007.05.025
Vile GF, Tanew-Ilitschew A, Tyrrell RM (1995) Activation of NF-kappa B in human skin fibroblasts by the oxidative stress generated by UVA radiation. Photochem Photobiol 62(3):463–468. https://doi.org/10.1111/j.1751-1097.1995.tb02369.x
Vincent SR (2010) Nitric oxide neurons and neurotransmission. Prog Neurobiol 90(2):246–255. https://doi.org/10.1016/j.pneurobio.2009.10.007
Westermann BA, Meissl H (2008) Nitric oxide synthase in photoreceptive pineal organs of fish. Comp Biochem Physiol A 151(2):198–204. https://doi.org/10.1016/j.cbpa.2008.06.032
Zaccone G, Mauceri A, Fasulo S (2006) Neuropeptides and nitric oxide synthase in the gill and the air-breathing organs of fishes. J Exp Zool 305(5):428–439. https://doi.org/10.1002/jez.a.267
Zhang H, Li B (2016) Pathological changes and iNOS expression in portal vein and hepatic artery of patients with liver cirrhosis and portal hypertension. Int J Clin Exp Pathol 9(12):12188–12196
Zhang C, Li DL, Chi C, Ling F, Wang GX (2015) Dactylogyrus intermedius parasitism enhances Flavobacterium columnare invasion and alters immune-related gene expression in Carassius auratus. Dis Aquat Org 116(1):11–21. https://doi.org/10.3354/dao02902
Acknowledgements
This study was supported by projects sanctioned to N. S. by the Science and Engineering Research Board, Department of Science and Technology, New Delhi, National Agricultural Science Fund, Indian Council of Agricultural Research, New Delhi, and the University Grants Commission, New Delhi. Rajiv Gandhi National Fellowship received by S.K is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
All animal experimentations were approved by the Animal Ethics Committee of North-Eastern Hill University, Shillong (No. IEC/NEHU/Ph.D./NOC/47). Furthermore, all experimental methods were performed according to the relevant guidelines and regulations of Animal Ethics Committee.
Rights and permissions
About this article
Cite this article
Kumari, S., Choudhury, M.G. & Saha, N. Hyper-ammonia stress causes induction of inducible nitric oxide synthase gene and more production of nitric oxide in air-breathing magur catfish, Clarias magur (Hamilton). Fish Physiol Biochem 45, 907–920 (2019). https://doi.org/10.1007/s10695-018-0593-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-018-0593-y