The Cerebellum

, Volume 10, Issue 3, pp 356–372

Spatially Restricted and Developmentally Dynamic Expression of Engrailed Genes in Multiple Cerebellar Cell Types

  • Sandra L. Wilson
  • Anna Kalinovsky
  • Grant D. Orvis
  • Alexandra L. Joyner
Article

Abstract

The cerebellum is a highly organized structure partitioned into lobules along the anterior–posterior (A-P) axis and into striped molecular domains along the medial–lateral (M-L) axis. The Engrailed (En) homeobox genes are required for patterning the morphological and molecular domains along both axes, as well as for the establishment of the normal afferent topography required to generate a fully functional cerebellum. As a means to understand how the En genes regulate multiple levels of cerebellum construction, we characterized En1 and En2 expression around birth and at postnatal day (P) 21 during the period when the cerebellum undergoes a remarkable transformation from a smooth ovoid structure to a highly foliated structure. We show that both En1 and En2 are expressed in many neuronal cell types in the cerebellum, and expression persists until at least P21. En1 and En2 expression, however, undergoes profound changes in their cellular and spatial distributions between embryonic stages and P21, and their expression domains become largely distinct. Comparison of the distribution of En-expressing Purkinje cells relative to early- and late-onset Purkinje cell M-L stripe proteins revealed that although En1- and En2-expressing Purkinje cell domains do not strictly align with those of ZEBRINII at P21, a clear pattern exists that is most evident at E17.5 by an inverse correlation between the level of En2 expression and PLCß4 and EPHA4.

Keywords

Engrailed Purkinje cell Cerebellum Compartment Development Stripe 

Supplementary material

12311_2011_254_Fig10_ESM.gif (194 kb)
Figure 1

Purkinje cells co-express LHX 1/5 and RORα. Representative sagittal (A-A″) and coronal (B-B″) sections of E17.5 cerebella show overlapping expression of LHX 1/5 (A and B) and RORα (A′ and B′). High magnification optical section taken from the dorsal Purkinje cell band displaying near complete overlap between LHX 1/5 (C and C″) and RORα (C′ and C″). Asterisk indicates a blood vessel fluorescing in the red channel (C and C″). (GIF 193 KB)

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High resolution image file (TIF 15.2 kb)
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Figure 2

EN1/2 is robustly expressed in ML interneurons, IGL and DCN cells, and at relatively lover levels in Purkinje cells at P21. (A) Nuclear labeling with pan-EN antibody (left panel, red in color composite) highlights most ML interneurons identified by immunodetection with parvalbumin (green), but not calbindin (blue) antibody. (B) Higher magnification of boxed region in (A). (C) All DCN subdivisions express EN1/2 at P21. (GIF 617 KB)

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High resolution image file (TIF 29.5 kb)
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Figure 3

Expression levels of EN1/2 in IGL at P21 differ between cerebellar subregions. Representative images of coronal series through P21 cerebellum demonstrating regional differences in EN expression levels in the IGL, with the highest expression in IGL concentrated in the nodular zone. (GIF 109 KB)

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High resolution image file (TIF 14.5 kb)

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Sandra L. Wilson
    • 1
  • Anna Kalinovsky
    • 1
  • Grant D. Orvis
    • 1
  • Alexandra L. Joyner
    • 1
  1. 1.Developmental Biology ProgramMemorial Sloan-Kettering Cancer CenterNew YorkUSA

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