Ex vivo culture of chick cerebellar slices and spatially targeted electroporation of granule cell precursors

Michalina Hanzel, Richard J T Wingate, Thomas Butts*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

The cerebellar external granule layer (EGL) is the site of the largest transit amplification in the developing brain, and an excellent model for studying neuronal proliferation and differentiation. In addition, evolutionary modifications of its proliferative capability have been responsible for the dramatic expansion of cerebellar size in the amniotes, making the cerebellum an excellent model for evo-devo studies of the vertebrate brain. The constituent cells of the EGL, cerebellar granule progenitors, also represent a significant cell of origin for medulloblastoma, the most prevalent paediatric neuronal tumour. Following transit amplification, granule precursors migrate radially into the internal granular layer of the cerebellum where they represent the largest neuronal population in the mature mammalian brain. In chick, the peak of EGL proliferation occurs towards the end of the second week of gestation. In order to target genetic modification to this layer at the peak of proliferation, we have developed a method for genetic manipulation through ex vivo electroporation of cerebellum slices from embryonic Day 14 chick embryos. This method recapitulates several important aspects of in vivo granule neuron development and will be useful in generating a thorough understanding of cerebellar granule cell proliferation and differentiation, and thus of cerebellum development, evolution and disease.

Original languageEnglish
Article numbere53421
JournalJournal of Visualized Experiments
Volume2015
Issue number106
DOIs
Publication statusPublished - 14 Dec 2015

Keywords

  • Cerebellum
  • Chick
  • Development
  • Developmental Biology
  • Electroporation
  • External granule layer
  • Granule cells
  • Issue 106
  • Purkinje cells
  • Slice culture

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