Cellular and subcellular imaging of motor protein-based behavior in embryonic rat brain

Methods Cell Biol. 2016:131:349-63. doi: 10.1016/bs.mcb.2015.06.013. Epub 2015 Sep 2.

Abstract

Development of the cerebral cortex is a very dynamic process, involving a series of complex morphogenetic events. Following division of progenitor cells in the ventricular zone, neurons undergo a series of morphological changes and migrate outward toward the cortical plate, where they differentiate and integrate into functional circuits. Errors at several of stages during neurogenesis and migration cause a variety of severe cortical malformations. A number of disease genes encode factors associated with the cytoskeleton, which plays a crucial role throughout cortical development. Methods for regulating gene expression coupled with imaging of subcellular structures have provided important insight into the mechanisms governing normal and abnormal brain development. We describe here a series of protocols for imaging motor protein-dependent processes in real time in the developing rat brain.

Keywords: Brain development; In utero electroporation; Interkinetic nuclear migration; Live imaging; Motor proteins; Neuronal migration; Radial glial progenitors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Movement / physiology
  • Cerebral Cortex / cytology
  • Cerebral Cortex / metabolism*
  • Electroporation / methods
  • Embryo, Mammalian / innervation
  • Ependymoglial Cells / cytology
  • Gene Expression Regulation / genetics
  • Green Fluorescent Proteins / genetics
  • Luminescent Proteins / genetics
  • Microtubules / metabolism
  • Molecular Motor Proteins / genetics*
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism*
  • Protein Transport / physiology
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Red Fluorescent Protein

Substances

  • Luminescent Proteins
  • Molecular Motor Proteins
  • RNA, Small Interfering
  • Green Fluorescent Proteins