Skip to main content

Advertisement

SpringerLink
Log in

Developmental exposure to acetaminophen does not induce hyperactivity in zebrafish larvae

  • Psychiatry and Preclinical Psychiatric Studies - Original Article
  • Published:
Journal of Neural Transmission Aims and scope Submit manuscript

Abstract

First line pain relief medication during pregnancy relies nearly entirely on the over-the-counter analgesic acetaminophen, which is generally considered safe to use during gestation. However, recent epidemiological studies suggest a risk of developing attention-deficit/hyperactivity disorder (ADHD)-like symptoms in children if mothers use acetaminophen during pregnancy. Currently, there are no experimental proofs that prenatal acetaminophen exposure causes developmental brain alterations of progeny. Exposure to high acetaminophen concentrations causes liver toxicity, which is well investigated in different model organisms. However, sub-liver-toxic concentrations have not been experimentally investigated with respect to ADHD endophenotypes such as hyperactivity. We used zebrafish to investigate the potential impact of acetaminophen exposure on locomotor activity levels, and compared it to the established zebrafish Latrophilin 3 (Lphn3) ADHD-model. We determined the sub-liver-toxic concentration of acetaminophen in zebrafish larvae and treated wild-type and lphn3.1 knockdown larvae with increasing concentrations of acetaminophen. We were able to confirm that lphn3.1 knockdown alone causes hyperactivity, strengthening the implication of Lphn3 dysfunction as an ADHD risk factor. Neither acute nor chronic exposure to acetaminophen at sub-liver-toxic concentrations in wild-type or lphn3.1 knock-downs increases locomotor activity levels. Together our findings show that embryonic to larval exposure to acetaminophen does not cause hyperactivity in zebrafish larvae. Furthermore, there are no additive and/or synergistic effects of acetaminophen exposure in a susceptible background induced by knock-down of lphn3.1. Our experimental study suggests that there is, at least in zebrafish larvae, no direct link between embryonic acetaminophen exposure and hyperactivity. Further work is necessary to clarify this issue in humans.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Arcos-Burgos M, Jain M, Acosta MT et al (2010) A common variant of the latrophilin 3 gene, LPHN3, confers susceptibility to ADHD and predicts effectiveness of stimulant medication. Mol Psychiatry 15:1053–1066

    Article  CAS  PubMed  Google Scholar 

  • Banerjee TD, Middelton F, Faraone SV (2007) Environmental risk factors for attention-deficit hyperactivity disorder. Acta Paediatr 96:1269–1274

    Article  PubMed  Google Scholar 

  • Brandlistuen RE, Ystrom E, Nulman I, Koren G, Nordeng H (2013) Prenatal paracetamol exposure and child neurodevelopment: a sibling-controlled cohort study. Int J Epidemiol 42:1702–1713

    Article  PubMed  PubMed Central  Google Scholar 

  • Driessen M, Kienhuis AS, Vitins AP, Pennings JLA, Pronk TE, van den Brandhof EJ, Roodbergen M, van de Water B, van der Ven LTM (2014) Gene expression markers in the zebrafish embryo reflect a hepatotoxic response in animal models and humans. Toxicol Lett 230:48–56

    Article  CAS  PubMed  Google Scholar 

  • Driessen M, Vitins AP, Pennings JLA, Kienhuis AS, van de Water B, van der Ven LTM (2015) A transcriptomics-based hepatotoxicity comparison between the zebrafish embryo and established human and rodent in vitro and in vivo models using cyclosporine a, amiodarone and acetaminophen. Toxicol Lett 232:403–412

    Article  CAS  PubMed  Google Scholar 

  • Franke B, Farone SV, Asheron P, Buitelaar J, Bau CHD, Ramos-Quiroga JA, Mick E, Grevet EH, Johansson S, Haavik J, Lesch K-P, Cormand B, Reif A (2012) The genetics of attention deficit/hyperactivity disorder in adults, a review. Mol Psychiatry 17:960–987

    Article  CAS  PubMed  Google Scholar 

  • Hawi Z, Cummins TDR, Tong J, Johnson B, Lau R, Samarrai W, Bellgrove MA (2015) The molecular genetic architecture of attention deficit hyperactivity disorder. Mol Psychiatry 20:289–297

    Article  CAS  PubMed  Google Scholar 

  • He JH, Guo SY, Zhu F, Zhu JJ, Chen YX, Huang CJ, Gao JM, Dong QX, Xuan YX, Li CQ (2013) A zebrafish phenotypic assay for assessing drug-induced hepatotoxicity. J Pharmacol Toxicol 67:25–32

    Article  CAS  Google Scholar 

  • Jaeschke H, Xie Y, McGill MR (2014) Acetaminophen-induced liver injury: from animal models to humans. J Clin Transl Hepatol 2(3):153–161

    PubMed  PubMed Central  Google Scholar 

  • Lange M, Norton W, Coolen M, Chaminade M, Merker S, Proft F, Schmitt A, Vernier P, Lesch K-P, Bally-Cuif L (2012) The ADHD-susceptibility gene lphn3.1 modulates dopaminergic neuron formation and locomotor activity during zebrafish development. Mol Psychiatry 17:946–954

    Article  CAS  PubMed  Google Scholar 

  • Liew Z, Ritz B, Rebordosa C, Lee P, Olsen J (2014) Acetaminophen use during pregnancy, behavioral problems, and hyperkinetic disorders. JAMA Pediatr 24:E1–E8

    Google Scholar 

  • Stewart AM, Braubach O, Spitsbergen J, Gerlai R, Kalueff AV (2014a) Zebrafish models for translational neuroscience research: from tank to bedside. Trends Neurosci 37:264–278

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stewart AM, Ullmann JFP, Norton WHJ, Parker MO, Brennan CH, Gerlai R, Kalueff AV (2014b) Molecular psychiatry of zebrafish. Mol Psychiatry 20:2–17

    Article  PubMed  PubMed Central  Google Scholar 

  • Thisse C, Thisse B (2008) High-resolution in situ hybridization to whole-mount zebrafish embryos. Nat Protocols 3:59–69

    Article  CAS  PubMed  Google Scholar 

  • Thomas R, Sanders S, Doust J, Beller E, Glasziou P (2015) Prevalence of attention-deficit/hyperactivity disorder: a systematic review and meta-analysis. Pediatr 135:e994–e1001

    Article  Google Scholar 

  • Thompson JMD, Waldie KE, Wall CR, Murphy R, Mitchell EA, the ABC study group (2014) Associations between acetaminophen use during pregnancy and ADHD symptoms measured at ages 7 and 11 years. PLoS One 9:e108210

    Article  PubMed  PubMed Central  Google Scholar 

  • van der Voet M, Harich B, Franke B, Schenck A (2015) ADHD-associated dopamine transporter, latrophilin and neurofibromin share a dopamine-related locomotor signature in Drosophila. Mol Psychiatry. doi:10.1038/mp.2015.55

    PubMed  PubMed Central  Google Scholar 

  • Viberg H, Eriksson P, Gordh T, Fredriksson A (2014) Paracetamol (acetaminophen) administration during neonatal brain development affects cognitive function and alters its analgesic and anxiolytic response in adult male mice. Toxicol Sci 138:139–147

    Article  CAS  PubMed  Google Scholar 

  • Vliegenthart ADB, Tucker CS, Del Pozo J, Dear JW (2014) Zebrafish as model organisms for studying drug induced liver injury. Br J Clin Pharmacol 78:1217–1227

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wallis D, Hill DS, Mendez IA, Abbott LC, Finnell RH, Wellman PJ, Setlow B (2012) Initial characterization of mice null for Lphn3, a gene implicated in ADHD and addiction. Brain Res 1463:85–92

    Article  CAS  PubMed  Google Scholar 

  • Werler M, Mitchell A, Hernandez_Diaz S, Honein M (2005) Use of over-the-counter medications during pregnancy. Am J Obstet Gynecol 193:771–777

    Article  PubMed  Google Scholar 

  • Zhang X, Gong Z (2013) Fluorescent transgenic zebrafish Tg(nkx2.2a:mEGFP) provides a highly sensitive monitoring tool for neurotoxins. PLoS One 8:e55474

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang X, Li C, Gong Z (2014) Development of a convenient in vivo hepatotoxin assay using a transgenic zebrafish line with liver-specific DsRed expression. PLoS One 9:e91874

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

We are grateful to M. Schartl for generously providing access to the fish facility and lab infrastructure, and to E Klopocki for qPCR support. Furthermore, we are thankful to W. Norton and M. Lange for advises regarding the lphn3.1 morphant behavior phenotype, Y. Kikuchi for generously sharing in situ probes, and C. Stigloher and W. Norton for critical comments on the manuscript. CL is funded by the Program Chancengleichheit für Frauen in Forschung und Lehre from the Bayerische Staatsregierung and University of Würzburg, and by a grant of the German Excellence Initiative to the Graduate School of Life Sciences, University of Würzburg.

Author information

Authors and Affiliations

  1. Department of Physiological Chemistry, Biocenter, Am Hubland, University of Würzburg, 97074, Würzburg, Germany

    Isabel Reuter & Christina Lillesaar

  2. Department of Human Genetics, Biocenter, Am Hubland, University of Würzburg, 97074, Würzburg, Germany

    Sabine Knaup

  3. Department of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital of Würzburg, 97080, Würzburg, Germany

    Marcel Romanos & Carsten Drepper

  4. Division of Molecular Psychiatry, Department of Psychiatry, Psychosomatics and Psychotherapy, ADHD Clinical Research Network, Laboratory of Translational Neuroscience, University of Würzburg, 97080, Würzburg, Germany

    Isabel Reuter & Klaus-Peter Lesch

Authors
  1. Isabel Reuter
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sabine Knaup
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marcel Romanos
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Klaus-Peter Lesch
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carsten Drepper
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christina Lillesaar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Carsten Drepper or Christina Lillesaar.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Additional information

C. Drepper and C. Lillesaar both share last authorship.

Electronic supplementary material

Below is the link to the electronic supplementary material.

702_2016_1556_MOESM1_ESM.jpg

Supplementary material 1. Injection of lphn3.1 splice morpholino causes loss of lphn3.1 transcripts. Reverse transcription followed by nested PCR performed on total RNA extracted from a pool of uninjected controls, lphn3.1 morphants or mismatch controls at 48 hpf. Arrow indicates wild-type lphn3.1 PCR-product (748 bp) (JPEG 114 kB)

702_2016_1556_MOESM2_ESM.jpg

Supplementary material 2. Morpholino knock-down of lphn3.1 increases locomotor activity in 6 dpf zebrafish larvae. Analysis of total distance swum (A), duration (B), swim speed (C) and number of entries into (D) inactivity, slow or fast swimming during a 5 minutes interval in uninjected controls, mismatch controls or lphn3.1 morphants. Numbers on bars in A indicate numbers of analyzed individuals and are the same for A-D.Bars and error bars illustrate mean+SD. * p<0.05 (JPEG 383 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Reuter, I., Knaup, S., Romanos, M. et al. Developmental exposure to acetaminophen does not induce hyperactivity in zebrafish larvae. J Neural Transm 123, 841–848 (2016). https://doi.org/10.1007/s00702-016-1556-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00702-016-1556-z

Keywords

Search

Navigation