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High Content Screening of Mammalian Primary Cortical Neurons

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High Content Screening

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1683))

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Abstract

High Content Screening (HCS) can be used to analyze the morphology of neuronal primary cultures on a large scale. When used in the field of neuronal regeneration this approach allows the screening of hundreds or thousands of perturbagens, such as miRNAs, cDNAs, or compounds, for their ability to induce neuronal growth. One of the most important steps while designing these kinds of experiments is the choice of the correct neuronal model. Testing the correct neuronal type is critical to obtain results that are biologically significant and that can later be translated to a clinical setting. For example, if the goal is identifying possible therapies for Spinal Cord Injury (SCI), a challenging target is the neuronal projection from the motor cortex to the spinal cord, the corticospinal tract. Here, we describe the experimental protocols that can be used to produce primary cortical culture from young rat cortices, electroporate the neurons to study the effect of altered gene expression on neurite growth, and immunostain to measure neurite growth parameters.

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Acknowledgments

We thank current and former members of the LemBix laboratory for developing our HCS approaches and thank Hassan Al-Ali for providing Fig. 1. V.P.L. holds the Walter G. Ross Distinguished Chair in Developmental Neuroscience. This work was supported by grants from the James and Esther King Biomedical Research Program JEK09KW-05 (J.L.B.), the US Army W81XWH-05-1-0061 (V.P.L., J.L.B.), the NIH [HD057521 (V.P.L.), NS059866 (J.L.B.)], and the Buoniconti Fund.

Author information

Authors and Affiliations

  1. Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, 1095 NW 14th Terrace (R-48), Miami, FL, 33136, USA

    Dario Motti Ph.D., John L. Bixby Ph.D. & Vance P. Lemmon Ph.D.

  2. Department of Biomedical Sciences, Marquette University, Milwaukee, WI, 53233, USA

    Murray Blackmore Ph.D.

Authors
  1. Dario Motti Ph.D.
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  2. Murray Blackmore Ph.D.
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  3. John L. Bixby Ph.D.
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  4. Vance P. Lemmon Ph.D.
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Corresponding author

Correspondence to Vance P. Lemmon Ph.D. .

Editor information

Editors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    Paul A. Johnston

  2. Bahama Bio, LLC, Bahama, North Carolina, USA

    Oscar J. Trask

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Motti, D., Blackmore, M., Bixby, J.L., Lemmon, V.P. (2018). High Content Screening of Mammalian Primary Cortical Neurons. In: Johnston, P., Trask, O. (eds) High Content Screening. Methods in Molecular Biology, vol 1683. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7357-6_17

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  • DOI: https://doi.org/10.1007/978-1-4939-7357-6_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7355-2

  • Online ISBN: 978-1-4939-7357-6

  • eBook Packages: Springer Protocols

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