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MicroRNA Technologies pp 29–42Cite as

Analysis of MicroRNAs and their Potential Targets in Human Embryonic Stem Cell-Derived Neurons Treated with the Anesthetic Propofol

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Part of the book series: Neuromethods ((NM,volume 128))

Abstract

Growing evidence demonstrates that prolonged exposure to general anesthetics, including propofol, induces widespread neuroapoptosis followed by long-term memory and learning disabilities in animal models. The underlying mechanisms of anesthetic-induced neurotoxicity are complex and not well understood. In addition, there is no direct clinical evidence of the detrimental effects of anesthetics in human fetuses, infants, or children. Development of an in vitro neurogenesis system of human stem cells opens up avenues of research for advancing our understanding of the issues of anesthetic-induced developmental neurotoxicity using a relevant human model. One avenue for investigating the mechanisms behind this neuroapoptosis is through evaluation of microRNA expression. MicroRNAs are endogenous, small, noncoding RNAs that negatively regulate target gene expression. MicroRNAs have been implicated to play important roles in many different disease processes, including neurological diseases. Our recent publication showed that among 84 microRNAs screened, propofol exposure altered the expression of 20 microRNAs in human embryonic stem cell (hESC)-derived neurons. Specifically, downregulation of microRNA-21 (miR-21) conferred by propofol played functional roles in the propofol-induced neurotoxicity. In this chapter, we outline (1) the protocol of human neuron differentiation from stem cells and (2) the protocols for analyzing microRNA expression (using our miR-21 study as an example) by quantitative reverse transcription-PCR and screening potential targets of miR-21 by Western blot assay following propofol exposure in stem cell-derived human neurons.

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Acknowledgement

This work was supported by R01GM112696 from the NIH (to Dr. Xiaowen Bai), by P01GM066730 and R01HL034708 from the NIH, Bethesda, MD, and by FP00003109 from Advancing a Healthier Wisconsin Research and Education Initiative Fund (to Dr. Zeljko J. Bosnjak).

Author information

Authors and Affiliations

  1. Department of Anesthesiology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA

    Danielle Twaroski, Jessica M. Olson & Zeljko J. Bosnjak

  2. Department of Physiology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA

    Danielle Twaroski, Jessica M. Olson, Mingyu Liang & Zeljko J. Bosnjak

  3. Departments of Anesthesiology and Physiology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA

    Yasheng Yan & Xiaowen Bai

Authors
  1. Danielle Twaroski
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  2. Yasheng Yan
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  3. Jessica M. Olson
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  4. Mingyu Liang
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  5. Zeljko J. Bosnjak
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  6. Xiaowen Bai
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Corresponding author

Correspondence to Xiaowen Bai .

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Editors and Affiliations

  1. Institute of Human Genetics, University of Cologne, Cologne, Germany

    Min Jeong Kye

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Twaroski, D., Yan, Y., Olson, J.M., Liang, M., Bosnjak, Z.J., Bai, X. (2016). Analysis of MicroRNAs and their Potential Targets in Human Embryonic Stem Cell-Derived Neurons Treated with the Anesthetic Propofol. In: Kye, M. (eds) MicroRNA Technologies. Neuromethods, vol 128. Humana Press, New York, NY. https://doi.org/10.1007/7657_2016_2

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  • DOI: https://doi.org/10.1007/7657_2016_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7173-2

  • Online ISBN: 978-1-4939-7175-6

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