Identifying domains of EFHC1 involved in ciliary localization, ciliogenesis, and the regulation of Wnt signaling

Ying Zhao, Jianli Shi, Mark Winey, Michael W. Klymkowsky

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

EFHC1 encodes a ciliary protein that has been linked to Juvenile Myoclonic Epilepsy. In ectodermal explants, derived from Xenopus laevis embryos, the morpholino-mediated down-regulation of EFHC1b inhibited multiciliated cell formation. In those ciliated cells that did form, axoneme but not basal body formation was inhibited. EFHC1b morphant embryos displayed defects in central nervous system (CNS) and neural crest patterning that were rescued by a EFHC1b-GFP chimera. EFHC1b-GFP localized to ciliary axonemes in epidermal, gastrocoele roof plate, and neural tube cells. In X. laevis there is a link between Wnt signaling and multiciliated cell formation. While down-regulation of EFHC1b led to a ~2-fold increase in the activity of the β-catenin/Wnt-responsive TOPFLASH reporter, EFHC1b-GFP did not inhibit β-catenin activation of TOPFLASH. Wnt8a RNA levels were increased in EFHC1b morphant ectodermal explants and intact embryos, analyzed prior to the on-set of ciliogenesis. Rescue of the EFHC1b MO's ciliary axonemal phenotypes required the entire protein; in contrast, the EFHC1b morpholino's Wnt8a, CNS, and neural crest phenotypes were rescued by a truncated form of EFHC1b. The EFHC1b morpholino's Wnt8a phenotype was also rescued by the injection of RNAs encoding secreted Wnt inhibitors, suggesting that these phenotypes are due to effects on Wnt signaling, rather than the loss of cilia, an observation of potential relevance to understanding EFHC1's role in human neural development.

Original languageEnglish (US)
Pages (from-to)257-265
Number of pages9
JournalDevelopmental Biology
Volume411
Issue number2
DOIs
StatePublished - Mar 15 2016
Externally publishedYes

Fingerprint

Morpholinos
Axoneme
Phenotype
Catenins
Embryonic Structures
Neural Crest
Xenopus laevis
Down-Regulation
Central Nervous System
Juvenile Myoclonic Epilepsy
RNA
Basal Bodies
Gastrula
Neural Tube
Cilia
Human Development
Proteins
Observation
Injections

Keywords

  • Biological science: Cell biology
  • Ciliogenesis
  • Developmental biology
  • Myoclonic epilepsy
  • Wnt signaling
  • Xenopus embryo

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

Identifying domains of EFHC1 involved in ciliary localization, ciliogenesis, and the regulation of Wnt signaling. / Zhao, Ying; Shi, Jianli; Winey, Mark; Klymkowsky, Michael W.

In: Developmental Biology, Vol. 411, No. 2, 15.03.2016, p. 257-265.

Research output: Contribution to journalArticle

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