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Comparison of molecular marker expression in early zebrafish brain development following chronic ethanol or morpholino treatment

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Abstract

This study was undertaken to ascertain whether defined markers of early zebrafish brain development are affected by chronic ethanol exposure or morpholino knockdown of agrin, sonic hedgehog, retinoic acid, and fibroblast growth factors, four signaling molecules that are suggested to be ethanol sensitive. Zebrafish embryos were exposed to 2% ethanol from 6 to 24 hpf or injected with agrin, shha, aldh1a3, or fgf8a morpholinos. In situ hybridization was employed to analyze otx2, pax6a, epha4a, krx20, pax2a, fgf8a, wnt1, and eng2b expression during early brain development. Our results showed that pax6a mRNA expression was decreased in eye, forebrain, and hindbrain of both chronic ethanol exposed and select MO treatments. Epha4a expression in rhombomere R1 boundary was decreased in chronic ethanol exposure and aldh1a3 morphants, lost in fgf8a morphants, but largely unaffected in agrin and shha morphants. Ectopic pax6a and epha4a expression in midbrain was only found in fgf8a morphants. These results suggest that while chronic ethanol induces obvious morphological change in brain architecture, many molecular markers of these brain structures are relatively unaffected by ethanol exposure.

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Acknowledgements

The authors thank Ms. Shanta Mackinnon for zebrafish husbandry. This work was supported by NIH Grant U54 AA019765.

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Authors and Affiliations

  1. Julius L. Chambers Biomedical/Biotechnology Research Institute, Durham, NC, USA

    Chengjin Zhang, Oswald Boa-Amponsem & Gregory J. Cole

  2. Integrated Biosciences Program, Durham, NC, USA

    Oswald Boa-Amponsem

  3. Department of Biological and Biomedical Sciences, North Carolina Central University, Durham, NC, 27707, USA

    Gregory J. Cole

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  1. Chengjin Zhang
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  2. Oswald Boa-Amponsem
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Correspondence to Gregory J. Cole.

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Zhang, C., Boa-Amponsem, O. & Cole, G.J. Comparison of molecular marker expression in early zebrafish brain development following chronic ethanol or morpholino treatment. Exp Brain Res 235, 2413–2423 (2017). https://doi.org/10.1007/s00221-017-4977-5

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