Mitotic granule cell precursors undergo highly dynamic morphological transitions throughout the external germinal layer of the chick cerebellum

Sci Rep. 2019 Oct 23;9(1):15218. doi: 10.1038/s41598-019-51532-y.

Abstract

The developing cerebellum of amniotes is characterised by a unique, transient, secondary proliferation zone: the external germinal layer (EGL). The EGL is comprised solely of granule cell precursors, whose progeny migrate inwardly to form the internal granule cell layer. While a range of cell morphologies in the EGL has long been known, how they reflect the cells' differentiation status has previously only been inferred. Observations have suggested a deterministic maturation from outer to inner EGL that we wished to test experimentally. To do this, we electroporated granule cell precursors in chick with plasmids encoding fluorescent proteins and probed labelled cells with markers of both proliferation (phosphohistone H3) and differentiation (Axonin1/TAG1 and NeuroD1). We show that granule cell precursors can display a range of complex forms throughout the EGL while mitotically active. Overexpression of full length NeuroD1 within granule cell precursors does not abolish proliferation, but biases granule cells towards precocious differentiation, alters their migration path and results in a smaller and less foliated cerebellum. Our results show that granule cells show a greater flexibility in differentiation than previously assumed. We speculate that this allows the EGL to regulate its proliferative activity in response to overall patterns of cerebellar growth.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Avian Proteins / analysis
  • Basic Helix-Loop-Helix Transcription Factors / analysis
  • Cell Movement
  • Cell Proliferation
  • Cerebellum / cytology
  • Cerebellum / embryology*
  • Chick Embryo / embryology*
  • Chickens
  • Mitosis
  • Nerve Tissue Proteins / analysis
  • Neural Stem Cells / cytology*
  • Neurogenesis

Substances

  • Avian Proteins
  • Basic Helix-Loop-Helix Transcription Factors
  • Nerve Tissue Proteins
  • Neurogenic differentiation factor 1