Modulation of brain serotonin by benzyl butyl phthalate in Fundulus heteroclitus (mummichog)

  • A. M. Deegan
  • R. B. Steinhauer
  • Richard S. Feinn
  • Matthew C. Moeller
  • H. M. PylypiwJr.
  • M. Nabel
  • C. J. Kovelowski
  • L. A. E. KaplanEmail author


Endocrine-disrupting chemicals have been known to alter important animal behaviors by modulating serotonin (5-hydroxytryptamine, 5-HT) and dopamine. F. heteroclitus (mummichog) brain serotonin and dopamine levels were quantified by enzyme-linked immunosorbent assay (ELISA) following a 28-day exposure regimen involving daily doses of either 0.1 mg l−1 benzyl butyl phthalate (BBP) dissolved in acetone or acetone alone (0.1 mg l−1). No differences in mean brain mass or total protein homogenate were induced by exposure to the acetone vehicle or BBP in acetone. The acetone vehicle had no effect on dopamine, serotonin, or tyrosine hydroxylase levels, but acetone did decrease tryptophan hydroxylase levels (p = 0.011). Exposure to BBP in acetone decreased dopamine (p = 0.024), increased serotonin (p < 0.001), reduced tryptophan hydroxylase as compared to the acetone vehicle alone (p < 0.001), and had no significant effect on tyrosine hydroxylase levels. This study is the first to report modulation of F. heteroclitus brain serotonin and its enzyme tryptophan hydroxylase following sub-lethal exposure to BBP in an acetone vehicle. In addition, modulation of brain dopamine in F. heteroclitus, sans simultaneous modulation of tyrosine hydroxylase, was also observed. These findings support the use of F. heteroclitus for assessing sub-lethal BBP exposure.


Estuary Fundulus Plasticizers Serotonin Dopamine 



We thank the College of Arts and Sciences and School of Health Sciences at Quinnipiac University for support of this research through Faculty Research Grants. We also thank our colleagues in the Departments of Biological Sciences and Chemistry & Physical Sciences at Quinnipiac University for their support in terms of time, talent and space to conduct and analyze these experiments. We acknowledge and are grateful for support from the CT Audubon Coastal Center (Milford, Connecticut) for providing unlimited access to the fish collection site, the Community Foundation for Greater New Haven’s Q-River Fund for providing support to complete the chemical testing of all water samples, and T. Folks from Rocky Mountain Diagnostics for his assistance with the ELISA kits.


This study was funded by an internal granting process supported by Quinnipiac University, College of Arts and Sciences (CAS Grant-in-Aid 2017-2018) awarded to Lisa A. E. Kaplan, Ph.D.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study involving animals were in accordance with the ethical standards of Quinnipiac University, where the studies were conducted, and approved by Quinnipiac University’s IACUC. The research did not involve human participants.

Informed consent

Informed consent was obtained from all individual participants, specifically authors, for whom identifying information is included in this article.


  1. Abraham BJ (1985) Species profiles. Life histories and environmental requirements of coastal fishes and invertebrates (Mid-Atlantic). MUMMICHOG AND STRIPED KILLIFISH. U.S. Fish Wildlife Service Biol. Rep. 82 (11.40); U.S. Army Corps Eng. Tech. Rep. EL-82-4Google Scholar
  2. Benjamin S, Masai E, Kamimura N, Takahashi K, Anderson RC, Faisal PA (2017) Phthalates impact human health: Epidemiological evidences and plausible mechanism of action. J Hazard Mater 340:360–383CrossRefGoogle Scholar
  3. Bisesi JH, Bridges W, Klaine SJ (2014) Effects of the antidepressant venlafaxine on fish brain serotonin and predation behavior. Aquat Toxicol 148:130–138CrossRefGoogle Scholar
  4. Blount BC, Silva MJ, Caudill SP, Needham LL, Pirkle JL, Sampson EJ, Lucier GW, Jackson RJ, Brock JW (2000) Levels of seven urinary phthalate metabolites in a human reference population. Exp Health Perspect 108:979–982CrossRefGoogle Scholar
  5. Boyer EW, Shannon M (2005) The serotonin syndrome. New Engl J Med 352:1112–1120CrossRefGoogle Scholar
  6. Chikae M, Ikeda R, Hatano Y, Hasan Q, Morita Y, Tamiya E (2004) Effects of bis(2-ethylhexyl) phthalate, γ-hexachlorocyclohexane, and 17β-estradiol on the fry stage of medaka (Oryzias latipes). Environ Toxicol Pharmacol 18:9–12CrossRefGoogle Scholar
  7. Crum KP, Balouskus RG, Targett TE (2018) Growth and movements of mummichogs (Fundulus heteroclitus) along armored and vegetated estuarine shorelines. Estuaries Coasts 41:131–143CrossRefGoogle Scholar
  8. Duke AA, Bègue L, Bell R, Eisenlohr-Moul T (2013) Revisiting the serotonin–aggression relation in humans: a meta-analysis. Psychol Bull 139:1148–1172CrossRefGoogle Scholar
  9. Dzieweczynski TL, Hebert OL (2012) Fluoxetine alters behavioral consistency of aggression and courtship in male Siamese fighting fish, Betta splendens. Physiol Behav 107:92–97CrossRefGoogle Scholar
  10. Edwards DH, Kravitz EA (1997) Serotonin, social status and aggression. Curr Opin Neurobiol 7:812–819CrossRefGoogle Scholar
  11. Fatoki O, Vernon F (1990) Phthalate esters in rivers of the Greater Manchester area, UK. Sci Total Environ 95:227–232CrossRefGoogle Scholar
  12. Gallagher S (2010) Treating Parkinson’s disease: dopamine dysregulation syndrome and impulse control. Br J Neurosci Nurs 6:24–28CrossRefGoogle Scholar
  13. Giam CS, Chan H, Neff G, Atlas EL (1978) Phthalate ester plasticizers: a new class of marine pollutant. Science 199:419–421CrossRefGoogle Scholar
  14. Gledhill WE, Kaley RG, Adams WJ, Hicks O, Michael PR, Saeger VW (1980) An environmental safety assessment of butyl benzyl phthalate. Environ Sci Technol 14:301–305CrossRefGoogle Scholar
  15. Goetz CG (2007) Textbook of clinical neurology. Saunders Elsevier, PhiladelphiaGoogle Scholar
  16. Gomez-Hens A, Aguilar-Caballos M (2003) Social and economic interest in the control of phthalic acid esters. TrAC Trends Anal Chem 22:847–857CrossRefGoogle Scholar
  17. Hoare DJ, Ruxton GD, Godin JJ, Krause J (2000) The social organization of free‐ranging fish shoals. Oikos 89:546–554CrossRefGoogle Scholar
  18. Howes OD, Kapur S (2009) The dopamine hypothesis of schizophrenia: version III—the final common pathway. Schizophr Bull 35:549–562CrossRefGoogle Scholar
  19. Iqbal MM, Basil MJ, Kaplan J, Iqbal M (2012) Overview of serotonin syndrome. Ann Clin Psychiatry 24:310–318Google Scholar
  20. Jobling S, Reynolds T, White R, Parker MG, Sumpter JP (1995) A variety of environmentally persistent chemicals, including some phthalate plasticizers, are weakly estrogenic. Environ Health Perspect 103:582–587CrossRefGoogle Scholar
  21. Kane A, Salierno J, Brewer S (2005) Fish models in behavioral toxicology: automated techniques, updates and perspectives. Methods Aquat Toxicol 2:559–590Google Scholar
  22. Kaplan LAE, Nabel M, Van Cleef-Toedt K, Proffitt AR, Pylypiw Jr. HM (2013) Impact of benzyl butyl phthalate on shoaling behavior in Fundulus heteroclitus (mummichog) populations. Mar Environ Res 86:70–75CrossRefGoogle Scholar
  23. Katsikantami I, Sifakis S, Tzatzarakis MN, Vakonaki E, Kalantzi OI, Tsatsakis AM, Rizos AK (2016) A global assessment of phthalates burden and related links to health effects. Environ Int 97:212–236CrossRefGoogle Scholar
  24. Kavaliers M (1980) Social groupings and circadian activity of the killifish, Fundulus heteroclitus. Biol Bull 158:69–76CrossRefGoogle Scholar
  25. Kulig B, Alleva E, Bignami G, Cohn J, Cory-Slechta D, Landa V, O’Donoghue J, Peakall D (1996) Animal behavioral methods in neurotoxicity assessment: SGOMSEC joint report. Environ Health Perspect 104(Suppl 2):193–204CrossRefGoogle Scholar
  26. Luo SX, Huang EJ (2016) Dopaminergic neurons and brain reward pathways: from neurogenesis to circuit assembly. Am J Pathol 186:478–488CrossRefGoogle Scholar
  27. Lyche JL, Gutleb AC, Bergman Å, Eriksen GS, Murk AJ, Ropstad E, Saunders M, Skaare JU (2009) Reproductive and developmental toxicity of phthalates. J Toxicol Environ Health, Part B 12:225–249CrossRefGoogle Scholar
  28. Min A, Liu F, Yang X, Chen M (2014) Benzyl butyl phthalate exposure impairs learning and memory and attenuates neurotransmission and CREB phosphorylation in mice. Food Chem Toxicol 71:81–89CrossRefGoogle Scholar
  29. Muller CP, Joacobs BL, Muller C (2010) Handbook of the behavioral neurobiology of serotonin. San Diego, California, USA: Elsevier.Google Scholar
  30. Narvaes R, Martins de A, Rosa M (2014) Aggressive behavior and three neurotransmitters: dopamine, GABA, and serotonin—A review of the last 10 years. Psychol Neurosci 7:601–607CrossRefGoogle Scholar
  31. O’Connor B, Kovacs T, Gibbons S, Strang A (1999) Carbon dioxide in pulp and paper mill effluents from oxygen-activated sludge treatment plants as a potential source of distress and toxicity to fish. Water Qual Res J Can 35:189–200CrossRefGoogle Scholar
  32. O’Sullivan SS, Evans AH, Lees AJ (2009) Dopamine dysregulation syndrome. CNS Drugs 23:157–170CrossRefGoogle Scholar
  33. Ptacek R, Stefano GB, Weissenberger S, Akotia D, Raboch J, Papezova H, Domkarova L, Stepankova T, Goetz M (2016) Attention deficit hyperactivity disorder and disordered eating behaviors: links, risks, and challenges faced. Neuropsychiatr Dis Treat 12:571–579CrossRefGoogle Scholar
  34. Purves D, Williams S (2001) Neuroscience, 2nd edn. Sinauer Associates, Sunderland, MAGoogle Scholar
  35. Robinson EC (1991) Lack of neuropathological changes in rats after exposure to butyl benzyl phthalate. J Toxicol Environ Health, Part A Curr Issues 32:345–347CrossRefGoogle Scholar
  36. Rodgers GM, Ward JR, Askwith B, Morrell LJ (2011) Balancing the dilution and oddity effects: decisions depend on body size. PLoS ONE 6(7):e14819CrossRefGoogle Scholar
  37. Schoots AM, Meijer RC, Denucé JM (1983) Dopaminergic regulation of hatching in fish embryos. Dev Biol 100:59–63CrossRefGoogle Scholar
  38. Scott GR, Sloman KA (2004) The effects of environmental pollutants on complex fish behaviour: integrating behavioural and physiological indicators of toxicity. Aquat Toxicol 68:369–392CrossRefGoogle Scholar
  39. Smith C (1985) The Inland Fishes of New York State, New York State Department of Environmental Conservation. Albany, New York, NYGoogle Scholar
  40. Sonnenschein C, Soto A, Fernandez M, Olea N, Olea-Serrano M, Ruiz-Lopez M (1995) Development of a marker of estrogenic exposure in human serum. Clin Chem 41:1888–1895Google Scholar
  41. Soto AM, Sonnenschein C, Chung KL, Fernandez MF, Olea N, Serrano FO (1995) The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants. Environ Health Perspect 103(Suppl 7):113–122CrossRefGoogle Scholar
  42. Vogl C, Grillitsch B, Wytek R, Spieser OH, Scholz W (1999) Qualification of spontaneous undirected locomotor behavior of fish for sublethal toxicity testing. Part I. Variability of measurement parameters under general test conditions. Environ Toxicol Chem 18:2736–2742CrossRefGoogle Scholar
  43. Ward AJ, Duff AJ, Horsfall JS, Currie S (2008) Scents and scents-ability: pollution disrupts chemical social recognition and shoaling in fish. Proc: Biol Sci 275:101–105Google Scholar
  44. Weber DN, Spieler RE (1994) Behavioral mechanisms of metal toxicity in fishes. In: Malins DC, Ostrander GK (Eds.) Aquatic toxicology: molecular, biochemical, and cellular perspectives. CRC Press, Boca Raton, FL, pp 421–467Google Scholar
  45. Weinberger J, Klaper R (2014) Environmental concentrations of the selective serotonin reuptake inhibitor fluoxetine impact specific behaviors involved in reproduction, feeding and predator avoidance in the fish Pimephales promelas (fathead minnow). Aquat Toxicol 151:77–83CrossRefGoogle Scholar
  46. Weis P, Weis JS (1974) DDT causes changes in activity and schooling behavior in goldfish. Environ Res 7:68–74CrossRefGoogle Scholar
  47. Whitworth WR (1996) Freshwater fishes of Connecticut: State Geological and Natural History Survey of Connecticut, Dept. of Environmental ProtectionGoogle Scholar
  48. Wibe ÅE, Billing A, Rosenqvist G, Jenssen BM (2002) Butyl benzyl phthalate affects shoaling behavior and bottom-dwelling behavior in threespine stickleback. Environ Res 89:180–187CrossRefGoogle Scholar
  49. Wibe ÅE, Fjeld E, Rosenqvist G, Jenssen BM (2004) Postexposure effects of DDE and butylbenzylphthalate on feeding behavior in threespine stickleback. Ecotoxicol Environ Saf 57:213–219CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biological SciencesQuinnipiac UniversityHamdenUSA
  2. 2.Department of Biomedical SciencesQuinnipiac UniversityHamdenUSA
  3. 3.Frank H. Netter, MD - School of MedicineQuinnipiac UniversityNorth HavenUSA
  4. 4.Department of Chemistry and Physical SciencesQuinnipiac UniversityHamdenUSA
  5. 5.Department of Mathematics & Computer ScienceQuinnipiac UniversityHamdenUSA

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