Expression of the δ-protocadherin gene Pcdh19 in the developing mouse embryo

doi:10.1016/j.modgep.2006.03.001

Gene Expression Patterns

Volume 6, Issue 8, 2 October 2006, Pages 893-899

https://doi.org/10.1016/j.modgep.2006.03.001 Get rights and content

Abstract

Protocadherins constitute a large family of transmembrane proteins primarily involved in weak homophilic adhesion in the brain and several other tissues. In a screen for potential regulators of kidney development, we have identified Pcdh19, a poorly characterized member of the δ-protocadherin subfamily. Here, we report the spatio-temporal expression pattern of Pcdh19 during mouse embryonic development. In midgestation embryos, Pcdh19 mRNA was detected in the mesonephros and in the neuroepithelium of the forebrain and midbrain. At later stages, Pcdh19 was expressed in other neural tissues such as the neural retina, nasal epithelium and spinal cord, as well as in the collecting duct and differentiating nephrons of the metanephros, in the glandular stomach, the exocrine pancreas and the hair follicles. Hence, the Pcdh19 gene is developmentally regulated during mouse organogenesis and shows a unique expression profile among protocadherins.

Section snippets

Results and discussion

Protocadherins (Pcdh) form the largest of six (6) major cadherin subfamilies (Nollet et al., 2000). They are characterized by the presence of a large first exon coding for extracellular ectodomain repeats (typically six or seven), a transmembrane domain and for a part of the cytoplasmic domain (Frank and Kemler, 2002, Suzuki, 2000). At the cellular level, protocadherins are thought to act as weak homophilic adhesion molecules. Although most of the roughly 70 Pcdh genes are found in three

Mice

Pax2^GFP BAC transgene (#30) used to sort mesonephros-specific cells was described previously (Grote et al., 2006, Pfeffer et al., 2002). Embryos for gene expression analysis were generated by timed matings of wild-type CD-1 mice. Noon of the day of plug detection was considered as E0.5.

Microarray analysis

FACS sorting of Pax2(+) mesonephric cells, linear amplification and DNA microarray analysis were described previously (Bouchard et al., 2005, Grote et al., 2006).

In situ hybridization and immunohistochemistry

Dissected embryos were fixed at 4 °C for 4 h to

Acknowledgements

We thank Susanne Kaitna and the members of the Bouchard’s laboratory for critical reading of the manuscript and Fréderic Charron for expertise on neural expression. This work was supported by the Canadian Institutes for Health Research and The Terry Fox Foundation. M.B holds a Canada Research Chair in Kidney Disease.

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Consistent with potential roles in early neural development, Pcdh19 expression has been shown to be expressed in the developing as well as adult brain (Dibbens et al., 2008; Pederick et al., 2016; Lin et al., 2012). Pcdh19 expression is first detected in the neural stem and progenitor cells (NSPCs) of the neuroepithelium at embryonic day 9 (E9) in mouse, and is expressed throughout the neuroepithelium by the onset of neurogenesis at E12.5 (Gaitan and Bouchard, 2006). At E15.5, the peak period of neurogenesis in mouse, Pcdh19 is highly expressed in both the NSPCs of the ventricular and sub-ventricular zones, the key proliferative zones of cortical development (Pederick et al., 2016).
PCDH19-Girls Clustering Epilepsy (PCDH19-GCE) is a childhood epileptic encephalopathy characterised by a spectrum of neurodevelopmental problems. PCDH19-GCE is caused by heterozygous loss-of-function mutations in the X-chromosome gene, Protocadherin 19 (PCDH19) encoding a cell-cell adhesion molecule. Intriguingly, hemizygous males are generally unaffected. As PCDH19 is subjected to random X-inactivation, heterozygous females are comprised of a mosaic of cells expressing either the normal or mutant allele, which is thought to drive pathology. Despite being the second most prevalent monogeneic cause of epilepsy, little is known about the role of PCDH19 in brain development. In this study we show that PCDH19 is highly expressed in human neural stem and progenitor cells (NSPCs) and investigate its function in vitro in these cells of both mouse and human origin. Transcriptomic analysis of mouse NSPCs lacking Pcdh19 revealed changes to genes involved in regulation of neuronal differentiation, and we subsequently show that loss of Pcdh19 causes increased NSPC neurogenesis. We reprogramed human fibroblast cells harbouring a pathogenic PCDH19 mutation into human induced pluripotent stem cells (hiPSC) and employed neural differentiation of these to extend our studies into human NSPCs. As in mouse, loss of PCDH19 function caused increased neurogenesis, and furthermore, we show this is associated with a loss of human NSPC polarity. Overall our data suggests a conserved role for PCDH19 in regulating mammalian cortical neurogenesis and has implications for the pathogenesis of PCDH19-GCE. We propose that the difference in timing or “heterochrony” of neuronal cell production originating from PCDH19 wildtype and mutant NSPCs within the same individual may lead to downstream asynchronies and abnormalities in neuronal network formation, which in-part predispose the individual to network dysfunction and epileptic activity.
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Importantly, gene inactivation of either Nephronectin or alpha8 integrin gene (Itga8) results in metanephric kidney agenesis (Linton et al., 2007). Pcdh19 and Wfdc2 are potential nephric duct regulators based on their strong expression previously reported in this tissue (Gaitan and Bouchard, 2006; Hellstrom et al., 2003). Together, these results identify important regulators of pro/mesonephros development requiring Pax2 function for normal expression.
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Expression of the δ-protocadherin gene Pcdh19 in the developing mouse embryo

Abstract

Section snippets

Results and discussion

Mice

Microarray analysis

In situ hybridization and immunohistochemistry

Acknowledgements

Dev. Biol.

Differentiation

Curr. Opin. Cell Biol.

Biochem. Biophys. Res. Commun.

Dev. Cell

Dev. Biol.

Biochim. Biophys. Acta

J. Mol. Biol.

Exp. Cell Res.

Genomics

FEBS Lett.

Identification of Pax2-regulated genes by expression profiling of the mid-hindbrain organizer region

Development

Functional equivalence of the transcription factors Pax2 and Pax5 in mouse development

Development