Advertisement

Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Growth hormone transgenesis affects osmoregulation and energy metabolism in zebrafish (Danio rerio)

  • 903 Accesses

  • 13 Citations

Abstract

Growth hormone (GH) transgenic fish are at a critical step for possible approval for commercialization. Since this hormone is related to salinity tolerance in fish, our main goal was to verify whether the osmoregulatory capacity of the stenohaline zebrafish (Danio rerio) would be modified by GH-transgenesis. For this, we transferred GH-transgenic zebrafish (T) from freshwater to 11 ppt salinity and analyzed survival as well as relative changes in gene expression. Results show an increased mortality in T versus non-transgenic (NT) fish, suggesting an impaired mechanism of osmotic acclimation in T. The salinity effect on expression of genes related to osmoregulation, the somatotropic axis and energy metabolism was evaluated in gills and liver of T and NT. Genes coding for Na+, K+-ATPase, H+-ATPase, plasma carbonic anhydrase and cytosolic carbonic anhydrase were up-regulated in gills of transgenics in freshwater. The growth hormone receptor gene was down-regulated in gills and liver of both NT and T exposed to 11 ppt salinity, while insulin-like growth factor-1 was down-regulated in liver of NT and in gills of T exposed to 11 ppt salinity. In transgenics, all osmoregulation-related genes and the citrate synthase gene were down-regulated in gills of fish exposed to 11 ppt salinity, while lactate dehydrogenase expression was up-regulated in liver. Na+, K+-ATPase activity was higher in gills of T exposed to 11 ppt salinity as well as the whole body content of Na+. Increased ATP content was observed in gills of both NT and T exposed to 11 ppt salinity, being statistically higher in T than NT. Taking altogether, these findings support the hypothesis that GH-transgenesis increases Na+ import capacity and energetic demand, promoting an unfavorable osmotic and energetic physiological status and making this transgenic fish intolerant of hyperosmotic environments.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

References

  1. Ayoola SO, Idowu AA (2008) Biotechnology and species development in aquaculture. Afr J Biotechnol 7(25):4722–4725

  2. Boisen A, Amstrup J, Novak I, Grosell M (2003) Sodium and chloride transport in soft water and hard water acclimated zebrafish (Danio rerio). Biochim Biophys Acta 1618:207–218

  3. Bolton J, Kawauchi H, Hirano T (1987) Osmoregulatory actions of growth hormone in rainbow trout (Salmo gairdneri). J Endocrinol 112:63–68

  4. Bouef G, Payan P (2001) How should salinity in influence fish growth? Comp Biochem Physiol C 130:411–423

  5. Breton S (2001) The cellular physiology of carbonic anhydrases. J Pancreas 2:159–164

  6. Carlson L, Bharadwaj R, Bartlke A (1999) Oxidative stress in hypopituitary dwarf mice and in transgenic mice overexpressing human and bovine GH. Age 22:181–186

  7. Cook JT, McNiven MA, Sutterlin AM (2000) Metabolic rate of pre-smolt growth-enhanced transgenic Atlantic salmon Salmo salar. Aquaculture 188:33–45

  8. Craig PM, Wood CM, McClelland GB (2007) Gill membrane remodeling with soft-water acclimation in zebrafish (Danio rerio). Physiol Genomics 30(1):53–60

  9. Deane EE, Woo NY (2005) Cloning and characterization of sea bream Na+K+-ATPase α and β subunit genes: in vitro effects of hormones on transcriptional and translational expression. Biochem Biophys Res Commun 331:1229–1238

  10. Deane EE, Woo NY (2006) Molecular cloning of growth hormone from silver sea bream: effects of abiotic and biotic stress on transcriptional and translational expression. Biochem Biophys Res Commun 342:1077–1082

  11. Devlin RH, Yesaki TY, Biagi CA, Donaldson EM, Swanson P, Chan WK (1994) Extraordinary growth in salmon. Nature 371:209–210

  12. Devlin RH, Yesaki TY, Donaldson EM, Shao JD, Hew C-L (1999) Production of germline transgenic Pacific salmonids with dramatically increased growth performance. Can J Fish Aquat Sci 52:1376–1384

  13. Drennon K, Moriyama S, Kawauchi H, Small B, Silverstein J, Parhar I, Shepherd B (2003) Development of an enzyme-linked immunosorbent assay for the measurement of plasma growth hormone (GH) levels in channel catfish (Ictalurus punctatus): assessment of environmental salinity and GH secretogogues on plasma GH levels. Gen Comp Endocrinol 133(3):314–322

  14. Du SJ, Gong Z, Fletcher GL, Shears MA, King MJ, Idler DR, Hew CL (1992) Growth enhancement in transgenic Atlantic salmon by the use of an ‘all fish’ chimeric growth hormone construct. Biotechnology 10:176–181

  15. Eppler E, Berishvili G, Mazel P, Caelers A, Hwang G, Maclean N, Reinecke M (2010) Distinct organ-specific up- and down-regulation of IGF-I and IGF-II mRNA in various organs of a GH-overexpressing transgenic Nile tilapia. Transgenic Res 19:231–240

  16. Evans DH, Piermarini PM, Potts W (1999) Ionic transport in the fish gill epithelium. J Exp Zool 283:641–652

  17. 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 85:97–177

  18. Figueiredo MD, Lanes CF, Almeida DV, Marins LF (2007a) Improving the production of transgenic fish germlines: in vivo evaluation of mosaicism in zebrafish (Danio rerio) using a green fluorescent protein (GFP) and growth hormone cDNA transgene co-injection strategy. Genet Mol Biol 30:31–36

  19. Figueiredo MD, Lanes CF, Almeida DV, Proietti MC, Marins LF (2007b) The effect of GH overexpression on GHR and IGF-I gene regulation in different genotypes of GH-transgenic zebrafish. Comp Biochem Physiol Part D 2:228–233

  20. Fridovich I (2004) Mitochondria: are they the seat of senescence? Aging Cell 3:13–16

  21. Guan B, Hu W, Zhang T, Wang Y, Zhu Z (2008) Metabolism traits of “all-fish” growth hormone transgenic common carp (Cyprinus carpio L.). Aquaculture 284:217–223

  22. Hallerman EM, McLean E, Fleming IA (2007) Effects of growth hormone transgenes on the behavior and welfare of aquacultured fishes: a review identifying research needs. Appl Animal Behav Sci 104:265–294

  23. Hamilton MA, Russo RC, Thurston RV (1977) Trimmed Spearman-Karber method for estimating median lethal concentrations in toxicity bioassays. Environ Sci Technol 11:714–719

  24. Higgs DA, Sutton JN, Kim H, Oakes JD, Smith J, Biagi C, Rowshandeli M, Devlin RH (2009) Influence of dietary concentrations of protein, lipid and carbohydrate on growth, protein and energy utilization, body composition, and plasma titres of growth hormone and insulin-like growth factor-1 in non-transgenic and growth hormone transgenic coho salmon, Oncorhynchus kisutch (Walbaum). Aquaculture 286:127–137

  25. Hirose S, Kaneko T, Naito N, Takei Y (2003) Molecular biology of major components of chloride cells. Comp Biochem Physiol Part B Biochem Mol Biol 136:593–620

  26. Hoshijima K, Hirose S (2007) Expression of endocrine genes in zebrafish larvae in response to environmental salinity. J Endocrinol 193(3):481–491

  27. Hwang PP (2009) Ion uptake and acid secretion in zebrafish (Danio rerio). J Exp Biol 212:1745–1752

  28. Jonassen TM, Pittman K, Imsland AK (1997) Seawater acclimation of tilapia, Oreochromis spilurus spilurus Günter, fry and fingerlings. Aquac Res 28:205–214

  29. Laiz-Carrión R, Sangiao-Alvarellos S, Guzmán JM, Río MP, Soengas JL, Mancera JM (2005) Growth performance of gilthead sea bream Sparus aurata in different osmotic conditions: implications for osmoregulation and energy metabolism. Aquaculture 250:849–861

  30. Lee CG, Devlin RH, Farrell AP (2003) Swimming performance, oxygen consumption and excess post-exercise oxygen consumption in adult transgenic and ocean-ranched coho salmon. J Fish Biol 62:753–766

  31. Leggatt RA, Devlin RH, Farrell AP, Randall DJ (2003) Oxygen uptake of growth hormone transgenic coho salmon during starvation and feeding. J Fish Biol 62:1053–1066

  32. Leggatt RA, Raven PA, Mommsen TP, Sakhrani D, Higgs D, Devlin RH (2009) Growth hormone transgenesis influences carbohydrate, lipid and protein metabolism capacity for energy production in coho salmon (Oncorhynchus kisutch). Comp Biochem Physiol Part B 154:121–133

  33. Lindskog S (1997) Structure and mechanism of carbonic anhydrase. Pharmacol Ther 74:1–20

  34. Mancera JM, McCormick SD (1998) Evidence for growth hormone/insulin-like growth factor I axis regulation of seawater acclimation in the euryhaline teleost Fundulus heteroclitus. Gen Comp Endocrinol 111:103–112

  35. Mancera JM, McCormick SD (1999) Influence of cortisol, growth hormone, insulin-like growth factor I and 3,3,5-triiodo-L-thyronine on hypoosmoregulatory ability in the euryhaline teleost Fundulus heteroclitus. Fish Physiol Biochem 21:25–33

  36. Mancera JM, McCormick SD (2000) Rapid activation of gill Na+, K+-ATPase in the euryhaline teleost Fundulus heteroclitus. J Exp Zool 287:263–274

  37. Marins LF, Iyengar A, Maclean N, Levy JA, Sohm F (2002) Simultaneous overexpression of GH and STAT5b genes inhibits the STAT5 signalling pathway in tilapia (Oreochromis niloticus) embryos. Genet Mol Biol 25:293–298

  38. Marshall WS (2002) Na+, Cl, Ca2+ and Zn2+ transport by fish gills: retrospective review and prospective synthesis. Transport 283:264–283

  39. McCormick SD (1993) Methods for nonlethal gill biopsy and measurement of Na+, K+-ATPase activity. Can J Fish Aquat Sci 50:656–658

  40. McCormick SD (1996) Effects of growth hormone and insulin-like growth factor I on salinity tolerance and gill Na+ , K+-ATPase in Atlantic salmon (Salmo salar): interaction with cortisol. Gen Comp Endocrinol 101:3–11

  41. McCormick SD (2001) Endocrine control of osmoregulation in teleost fish. Am Zool 41:781–794

  42. Muradian KK, Utko NA, Mozzhukhina TG, Litoshenko AY, Pishel IN, Bezrukov VV, Fraifield VE (2002) Pair-wise linear and 3d nonlinear relationships between the liver antioxidant enzyme activities and the rate of body oxygen consumption in mice. Free Radical Biol Med 33:1736–1739

  43. Nam YK, Noh JK, Cho YS, Cho HJ, Cho KN, Kim CG, Kim DS (2001) Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis. Transgenic Res 10:353–362

  44. Parks SK, Tresguerres M, Goss GG (2008) Review of theoretical considerations underlying Na+ uptake mechanisms in freshwater fishes. Comp Biochem Physiol Part C 148:411–418

  45. Perry SF, Shahsavarani A, Georgalis T, Bayaa M, Furimsky M, Thomas SL (2003) Channels, pumps, and exchangers in the gill and kidney of freshwater fishes: their role in ionic and acid–base regulation. J Exp Zool Part A Comp Exp Biol 300:53–62

  46. Peterson MS, Comyns BH, Rakocinski CF, Fulling GL (1999) Does salinity affect somatic growth in early juvenile Atlantic croaker, Micropogonias undulatus (L.)? J Exp Mar Biol Ecol 238:199–207

  47. Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30:e36

  48. Pitkänen TI, Krasnov A, Teerijoki H, Mölsä H (1999) Transfer of growth hormone (GH) transgenes into Arctic charr (Salvelinus alpinus L.) I. Growth response to various GH constructs. Genet Anal Biomol Eng 15:91–98

  49. Rahman MA, Mak R, Ayad H, Smith A, Maclean N (1998) Expression of a novel piscine growth hormone gene results in growth enhancement in transgenic tilapia (Oreochromis niloticus). Transg Res 7:357–369

  50. Reinecke M (2010) Influences of the environment on the endocrine and paracrine fish growth hormone–insulin-like growth factor-I system. J Fish Biol 76:1233–1254

  51. Rosa CE, Figueiredo MA, Almeida DV, Marins LF (2008) Metabolic rate and reactive oxygen species production in different genotypes of GH-transgenic zebrafish. Comparative Biochemistry and PhysiologyA. Mol Integr Physiol 149:209–214

  52. Sakamoto T, Hirano T (1993) Expression of insulin-like growth factor I gene in osmoregulatory organs during seawater adaptation of the salmonid fish: possible mode of osmoregulatory action of growth hormone. Proc Natl Acad Sci USA 90:1912–1916

  53. Sakamoto T, McCormick SD (2006) Prolactin and growth hormone in fish osmoregulation. Gen Comp Endocrinol 147:24–30

  54. Sakamoto T, Shepherd BS, Madsen SS, Nishioka RS, Siharath K, Richman NH, Bern HA, Grau EG (1997) Osmoregulatory actions of growth hormone and prolactin in an advanced teleost. Gen Comp Endocrinol 106:95–101

  55. Sangiao-Alvarellos S, Laiz-Carrión R, Guzmán JM, Martín Del Río MP, Miguez JM, Mancera JM, Soengas JL (2003) Acclimation of S. aurata to various salinities alters energy metabolism of osmoregulatory and nonosmoregulatory organs. Am J Physiol Reg Integr Comp Physiol 285:897–907

  56. Sangiao-Alvarellos S, Míguez JM, Soengas JL (2005) Actions of growth hormone on carbohydrate metabolism and osmoregulation of rainbow trout (Oncorhynchus mykiss). Gen Comp Endocrinol 141:214–225

  57. Seidelin M, Madsen SS, Byrialsen A, Kristiansen K (1999) Effects of insulin-like growth factor-I and cortisol on Na+ , K+-ATPase expression in osmoregulatory tissues of brown trout (Salmo trutta). Gen Comp Endocrinol 113:331–342

  58. Shrimpton J, Devlin RH, Mclean E, Byatt CJ, Donaldson E, Randall D (1995) Increases in gill cytosolic corticosteroid receptor abundance and saltwater tolerance in juvenile coho salmon (Oncorhynchus kisutch) treated with growth hormone and placental lactogen. Gen Comp Endocrinol 98:1–15

  59. Tang Y, Shepherd BS, Nichols AJ, Dunham R, Chen TT (2001) Influence of environmental salinity on messenger RNA levels of growth hormone, prolactin, and somatolactin in pituitary of the channel catfish (Ictalurus punctatus). Mar Biotechnol 3:205–217

  60. Tseng Y, Hwang P (2008) Some insights into energy metabolism for osmoregulation in fish. Comp Biochem Physiol Part C 148:419–429

  61. Tseng YC, Lee JR, Chang JCH, Kuo CH, Lee SJ, Hwang PP (2008) Regulation of lactate dehydrogenase in tilapia (Oreochromis mossambicus) gills during acclimation to salinity challenge. Zoolog Stud 47:473–480

  62. Wheatly BM, Henry RP (1987) Branchial and antennal gland Na+/K+-dependent ATPase and carbonic anhydrase activity during salinity acclimation of the euryhaline crayfish Pacifastacus leniusculus. J Exp Biol 133:73–86

  63. Xu B, Miao H, Zhang P, Li D (1997) Osmoregulatory actions of growth hormone in juvenile tilapia (Oreochromis niloticus). Fish Physiol Biochem 17:295–301

  64. Yan JJ, Chou MY, Kaneko T, Hwang PP (2007) Gene expression of Na+/H+ exchanger in zebrafish H+ -ATPase-rich cells during acclimation to low-Na+ and acidic environments. Am J Physiol 293:1814–1823

Download references

Acknowledgments

The authors would like to thank Maíra Proietti for valuable corrections of the manuscript. This work was supported by Brazilian CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior). AB is a research fellow from CNPq (Proc. # 304430/2009-9) and is supported by the International Canada Research Chair Program from the International Development Research Centre (Ottawa, Canada).

Author information

Correspondence to Luis Fernando Marins.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Almeida, D.V., de Martinez Gaspar Martins, C., de Azevedo Figueiredo, M. et al. Growth hormone transgenesis affects osmoregulation and energy metabolism in zebrafish (Danio rerio). Transgenic Res 22, 75–88 (2013). https://doi.org/10.1007/s11248-012-9627-x

Download citation

Keywords

  • Energy metabolism
  • Gene expression
  • Growth hormone
  • Somatrotopic axis
  • Salinity
  • Transgenic zebrafish