High content screening of mammalian primary cortical neurons

Dario Motti, Murray Blackmore, John Bixby, Vance Lemmon

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages293-304
Number of pages12
Volume1683
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1683
ISSN (Print)1064-3745

Fingerprint

Neurites
Neurons
Growth
Pyramidal Tracts
Motor Cortex
MicroRNAs
Spinal Cord Injuries
Regeneration
Spinal Cord
Complementary DNA
Gene Expression
Therapeutics
caN protocol

Keywords

  • Electroporation
  • Phenotypic screening
  • Primary cortical neuron
  • Rat

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Motti, D., Blackmore, M., Bixby, J., & Lemmon, V. (2018). High content screening of mammalian primary cortical neurons. In Methods in Molecular Biology (Vol. 1683, pp. 293-304). (Methods in Molecular Biology; Vol. 1683). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7357-6_17

High content screening of mammalian primary cortical neurons. / Motti, Dario; Blackmore, Murray; Bixby, John; Lemmon, Vance.

Methods in Molecular Biology. Vol. 1683 Humana Press Inc., 2018. p. 293-304 (Methods in Molecular Biology; Vol. 1683).

Research output: Chapter in Book/Report/Conference proceedingChapter

Motti, D, Blackmore, M, Bixby, J & Lemmon, V 2018, High content screening of mammalian primary cortical neurons. in Methods in Molecular Biology. vol. 1683, Methods in Molecular Biology, vol. 1683, Humana Press Inc., pp. 293-304. https://doi.org/10.1007/978-1-4939-7357-6_17
Motti D, Blackmore M, Bixby J, Lemmon V. High content screening of mammalian primary cortical neurons. In Methods in Molecular Biology. Vol. 1683. Humana Press Inc. 2018. p. 293-304. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7357-6_17
Motti, Dario ; Blackmore, Murray ; Bixby, John ; Lemmon, Vance. / High content screening of mammalian primary cortical neurons. Methods in Molecular Biology. Vol. 1683 Humana Press Inc., 2018. pp. 293-304 (Methods in Molecular Biology).
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