ABSTRACT
Wnt signaling plays an important role in cell growth, differentiation, polarity formation, and neural development. We have recently identified the Coiled-coil-DIX1 (Ccd1) gene encoding a third type of a DIX domain-containing protein. Ccd1 forms homomeric and heteromeric complexes with Dishevelled and Axin, and positively regulates the Wnt/beta-catenin pathway. Here, we examined the spatiotemporal expression pattern of Ccd1 mRNA in mouse embryos from embryonic day 6.5 (E6.5) to E17.5 by in situ hybridization. Ccd1 expression was detected in the node region in gastrula embryos, in the cephalic mesenchyme and tail bud at E8.5, and in the branchial arch and forelimb bud at E9.5. In the central nervous system, Ccd1 expression began and persisted in the regions where the neurons differentiated, so that it was observed throughout the brain and spinal cord at E17.5. Ccd1 expression was also strong in the peripheral nervous system, including sensory cranial ganglia (trigeminal, facial, and vestibulocochlear ganglia), dorsal root ganglia, and autonomic ganglia (sympathetic ganglia, celiac ganglion, and hypogastric plexus). Ccd1 was detected in the sensory organs, such as the inner nuclear layer of the neural retina, saccule and cochlea of the inner ear, and nasal epithelium. Outside the nervous system, Ccd1 mRNA was observed in the cartilage, tongue, lung bud, stomach, and gonad at E12.5-E14.5, and in the tooth bud, bronchial epithelium, and kidney at E17.5. Taken together, these findings demonstrate that Ccd1 expression is observed in all the neurons in the nervous system, closely associated with neural crest-derived tissues, and largely overlapping with the regions where several Wnt genes are reported to play a role.
Subject(s)
Embryonic Development , Gene Expression Regulation, Developmental , Proteins/metabolism , Signal Transduction , Wnt Proteins/metabolism , Animals , Central Nervous System/metabolism , In Situ Hybridization , Intracellular Signaling Peptides and Proteins/metabolism , Mice , Mice, Inbred C57BL , Neurons/metabolism , Peripheral Nervous System/metabolism , RNA, Messenger/metabolism , Tissue DistributionABSTRACT
The Wnt signaling plays important roles in cell growth, differentiation, polarity formation, and neural development. In the canonical pathway, two DIX domain-containing proteins, Dishevelled (Dvl) and Axin, regulate the degradation of beta-catenin that activates Wnt target genes through TCF/LEF family transcription factors. Recently, we have isolated a third type of DIX domain-possessing protein, Coiled-coil-DIX1 (Ccd1). Ccd1 forms homomeric and heteromeric complexes with Dvl and Axin, and regulates the neural patterning in zebrafish embryos through Wnt pathway activation. Here, we report the isolation and characterization of mouse Ccd1. Fourteen putative mRNA isoforms are generated by different promoter usage and alternative splicing, and each isoform shows different expression patterns in various tissues. The predicted Ccd1 proteins are classified into three subtypes, and a novel form, termed Ccd1A, possesses an N-terminal calponin homology domain, suggesting an additional interaction of the isoform with actin or other proteins. When Ccd1 proteins were singularly expressed in Hela cells, they showed almost no activation of TCF-dependent reporter transcription on their own. However, when Dvl protein, at the level that did not activate Wnt pathway by itself, was co-expressed with Ccd1, the reporter transcription was greatly potentiated in Ccd1-dose-dependent manner. In addition, Ccd1- and Wnt3a-dependent activation of Wnt pathway was inhibited by Axin or a dominant negative Ccd1. These results indicate that mouse Ccd1 functions as a positive regulator of the Wnt/beta-catenin pathway. Furthermore, Ccd1 is highly expressed and co-localized with Wnt signaling molecules in the embryonic and adult brain, implicating the importance of Ccd1 in the Wnt-mediated neuronal development, plasticity, and remodeling.
