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Fluorescent protein transgenesis has varied effects on behaviour and cold tolerance in a tropical fish (Gymnocorymbus ternetzi): implications for risk assessment

  • Rosalind A. LeggattEmail author
  • Robert H. Devlin
Article

Abstract

Fluorescent protein (FP) transgenesis is used in the ornamental aquarium trade to produce new colour morphs in tropical fish. Understanding whether such genetic modification could alter ability to survive temperate waters, or interactions with native fish, should such fish be released to natural systems is critical in developing policy on their commercial use. We examined the competitive foraging ability and cold tolerance of unrelated pet-trade sourced adult green FP transgenic tetra and non-transgenic white tetra (Gymnocorymbus ternetzi), as well as white non-transgenic and green FP transgenic juvenile progeny of these groups. FP transgenesis did not affect the foraging success or aggressive behaviour in either adult or juvenile fish, indicating FP transgenesis may not influence potential hazards through this pathway. During a cold temperature tolerance trial, adult green tetras had greatly diminished cold tolerance relative to unrelated adult white fish, while sibling juvenile offspring of these groups had intermediate cold tolerance between adult fish groups that were not affected by FP transgenesis. This data suggests background genetics, rearing history and/or life stage may play larger roles in cold tolerance than FP transgenesis in this species. Unexpectedly, both adult and juvenile white tetras were 3.8 times more likely to take refuge in shelters when temperature declined than green tetras. These data indicate FP transgenic fish may pose equal or lesser risk than non-transgenic fish, should they be released to natural environments. Results also demonstrate that unrelated pet-trade sourced fish may not always be appropriate models for examining effects of FP transgenesis.

Keywords

Fluorescent protein Transgenic Tropical fish Environmental risk Cold tolerance Competition 

Notes

Acknowledgments

This project was funded by the Canadian Regulatory System for Biotechnology. Thanks go to A. Csuzdi, H. Tadey, and B. Yates for assistance in fish care and monitoring.

Compliance with ethical standards

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted (Pacific Region Animal Care Committee, AUP18-014).

Supplementary material

10695_2019_725_MOESM1_ESM.docx (1 mb)
ESM 1 (DOCX 1045 kb)

References

  1. Badrian B, Bogoyevitch MA (2007) Changes in the transcriptional profile of cardiac myocytes following green fluorescent protein expression. DNA Cell Biol 26:727–736.  https://doi.org/10.1089/dna.2007.0604 CrossRefPubMedGoogle Scholar
  2. Baens J et al (2006) The dark side of EGFP: Defective polyubiquitination. PLoS ONE 1:e54.  https://doi.org/10.1371/journal.pone.0000054 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Barrett RDH, Paccard A, Healy TM, Bergek S, Schulte PM, Schluter D, Rogers SM (2011) Rapid evolution of cold tolerance in stickleback. Proc R Soc Lond B Biol Sci 278:233–238.  https://doi.org/10.1098/rspb.2010.0923 CrossRefGoogle Scholar
  4. Charo-Karisa H, Rezk MA, Bovenhuis H, Komen H (2005) Heritability of cold tolerance in Nile tilapia, Oreochromis niloticus, juveniles. Aquaculture 249:115–123.  https://doi.org/10.1016/j.aquaculture.2005.04.029 CrossRefGoogle Scholar
  5. Core Team R (2018) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria Available from https://www.r-project.org/.Google Scholar
  6. Cortemeglia C, Beitinger TL (2005) Temperature tolerances of wild-type and red transgenic zebra danios. Trans Am Fish Soc 134:1431–1437.  https://doi.org/10.1577/T04-197.1 CrossRefGoogle Scholar
  7. Cortemeglia C, Beitinger TL (2006a) Projected US distributions of transgenic and wildtype zebra danios, Danio rerio, based on temperature tolerance data. J Therm Biol 31:422–428.  https://doi.org/10.1016/j.jtherbio.2006.01.011 CrossRefGoogle Scholar
  8. Cortemeglia C, Beitinger TL (2006b) Susceptibility of transgenic and wildtype zebra danios, Danio rerio, to predation. Environ Biol Fishes 76:93–100.  https://doi.org/10.1007/s10641-006-9011-x CrossRefGoogle Scholar
  9. Coumans JVF, Gau D, Polijak A, Wasinger V, Roy P, Moens PR (2014) Green fluorescent protein expression triggers proteome changes in breast cancer cells. Exp Cell Res 320:33–45.  https://doi.org/10.1016/j.yexcr.2013.07.019 CrossRefPubMedGoogle Scholar
  10. Devlin RH, D’Adrade M, Uh M, Biagi CA (2004) Population effects of growth hormone transgenic coho salmon depend on food availability and genotype by environment interactions. Proc Natl Acad Sci U S A 101:9303–9308.  https://doi.org/10.1073/pnas.0400023101 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Devlin RH, Sundstrom LF, Leggatt RA (2015) Assessing ecological and evolutionary consequences of growth-accelerated genetically engineered fishes. Bioscience 65:685–700.  https://doi.org/10.1093/biosci/biv068 CrossRefGoogle Scholar
  12. DFO (2018) Environmental and indirect human health risk assessment of the Glofish® Electric Green® Tetra and the Glofish® Long-Fin Electric Green® Tetra (Gymnocorymbus ternetzi): a transgenic ornamental fish. DFO Can Sci Advis Sec Sci Advis Rep 2018/027 23 pp. https://waves-vagues.dfo-mpo.gc.ca/Library/40712928.pdf
  13. DFO (2019) Environmental and indirect human health risk assessment of the GloFish® Tetras (Gymnocorymbus ternetzi): five lines of transgenic ornamental fish. DFO Can Sci Advis Sec Sci Advis Rep 2019/002 26 pp. www.dfo-mpo.gc.ca/csas-sccs/Publications/SAR-AS/2019/2019_002-eng.pdf
  14. Hill JE, Kapuscinski AR, Pavlowich T (2011) Fluorescent transgenic zebra danio more vulnerable to predators than wild-type fish. Trans Am Fish Soc 140:1001–1005.  https://doi.org/10.1080/00028487.2011.603980 CrossRefGoogle Scholar
  15. Howard RD, Rohrer K, Liu YY, Muir WM (2015) Mate competition and evolutionary outcomes in genetically modified zebrafish (Danio rerio). Evolution 69:1143–1157.  https://doi.org/10.1111/evo.12662 CrossRefPubMedGoogle Scholar
  16. Jha P (2010) Comparative study of aggressive behaviour in transgenic and wildtype zebrafish Danio rerio (Hamilton) and the flying barb Esomus danricus (Hamilton), and their susceptibility to predation by the snakehead Channa striatus (Bloch). Ital J Zool 77:102–109.  https://doi.org/10.1080/11250000802629463 CrossRefGoogle Scholar
  17. Leggatt RA (2019) Cold temperature tolerance of albino rainbow shark (Epalzeorhynchos frenatum), a tropical fish with transgenic application in the ornamental aquarium trade. Can J Zool 97:1–3.  https://doi.org/10.1139/cjz-2018-0208 CrossRefGoogle Scholar
  18. Leggatt RA, Dhillion RS, Mimeault C, Johnson N, Richards JG, Devlin RH (2018a) Low-temperature tolerances of tropical fish with potential transgenic applications in relation to winter water temperatures in Canada. Can J Zool 96:253–260.  https://doi.org/10.1139/cjz-2017-0043 CrossRefGoogle Scholar
  19. Leggatt RA, Johnson N, McGowan C (2018b) Environmental risk assessment of the Glofish® Electric Green® Tetra and the Glofish® Long-Fin Electric Green® Tetra: transgenic ornamental fish, imported to Canada, for sale in the pet trade. DFO Can Sci Advis Sec Res Doc 2018/049 xii + 54 pp. https://waves-vagues.dfo-mpo.gc.ca/Library/40722995.pdf
  20. Li H, Wei H, Wang Y, Tang H, Wang Y (2013) Enhanced green fluorescent protein transgenic expression in vivo is not biologically inert. J Proteome Res 12:3801–3808.  https://doi.org/10.1021/pr400567g CrossRefPubMedGoogle Scholar
  21. Mak GW-Y, Wong C-H, Tsui SK-W (2007) Green fluorescent protein induces the secretion of inflammatory cytokine interleukin-6 in muscle cells. Anal Biochem 362:296–298.  https://doi.org/10.1016/j.ab.2006.12.017 CrossRefPubMedGoogle Scholar
  22. Owen MA, Rohrer K, Howard RD (2012) Mate choice for a novel male phenotype in zebrafish, Danio rerio. Anim Behav 83:811–820.  https://doi.org/10.1016/j.anbehav.2011.12.029 CrossRefGoogle Scholar
  23. Saillant E, Wang XX, Ma L, Gatlin DM, Vega RR, Gold JR (2008) Genetic effects on tolerance to acute cold stress in red drum, Sciaenops ocellatus L. Aquac Res 39:1393–1398.  https://doi.org/10.1111/j.1365-2109.2008.02008.x CrossRefGoogle Scholar
  24. Schaefer J, Ryan A (2006) Developmental plasticity in the thermal tolerance of zebrafish Danio rerio. J Fish Biol 69:772–734.  https://doi.org/10.1111/j.1095-8649.2006.01145.x CrossRefGoogle Scholar
  25. Snekser JL, McRobert SP, Murphy CE, Clotfelter ED (2006) Aggregation behavior in wildtype and transgenic zebrafish. Ethology 112:181–187.  https://doi.org/10.1111/j.1439-0310.2006.01139.x CrossRefGoogle Scholar
  26. Stewart CN (2006) Go with the glow: fluorescent proteins to light transgenic organisms. Trends Biotechnol 24:155–162.  https://doi.org/10.1016/j.tibtech.2006.02.002 CrossRefPubMedGoogle Scholar
  27. Travis J, McManus MG, Baer CF (1999) Sources of variation in physiological phenotypes and their evolutionary significance. Am Zool 39:422–433.  https://doi.org/10.1093/icb/39.2.422 CrossRefGoogle Scholar
  28. Truebano M, Fenner P, Tills O, Rundle SD, Rezende EL (2018) Thermal strategies vary with life history stage. J Exp Biol 221:1–5.  https://doi.org/10.1242/jeb.171629 CrossRefGoogle Scholar
  29. Tuckett QM, Ritch JL, Lawson KM, Lawson LL, Hill JE (2016) Variation in cold tolerance in escaped and farmed non-native green swordtails (Xiphophorus hellerii) revealed by laboratory trials and field introductions. Biol Invasions 18:45–56.  https://doi.org/10.1007/s10530-015-0988-y CrossRefGoogle Scholar
  30. USFWS (US Fish & Wildlife Service) (2017) Black tetra (Gymnocorymbus ternetzi): ecological risk screening summary. Available at https://www.fws.gov/fisheries/ANS/erss/uncertainrisk/ERSS-Gymnocorymbus-ternetzi-FINAL-November2017.pdf. Accessed August 2019.
  31. Venables WN, Ripley BD (2002) Modern applied statistics with S. Springer, New YorkCrossRefGoogle Scholar

Copyright information

© © Crown 2019

Authors and Affiliations

  1. 1.Fisheries and Oceans CanadaWest VancouverCanada

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