Reconstitution of the Cytoplasmic Regulation of the Wnt Signaling Pathway Using Xenopus Egg Extracts

Abstract

The regulation of β-catenin turnover is the central mechanism governing activation of the Wnt signaling pathway. All components of the pathway are present in the early embryo of Xenopus laevis, and Xenopus egg extracts have been used to recapitulate complex biological reactions such as microtubule dynamics, DNA replication, chromatin assembly, and phases of the cell cycle. Herein, we describe a biochemical method for analyzing β-catenin degradation using radiolabeled and luciferase-fusion proteins in Xenopus egg extracts. We show that in such a biochemical system, cytoplasmic β-catenin degradation is regulated by soluble components of the Wnt pathway as well as small molecules.

Wnt/β-Catenin Signaling in Neural Stem Cell Homeostasis and Neurological Diseases

“…Wnt signaling plays a key role in the regulation of neurogenesis and tissue homeostasis in the nervous system (Logan and Nusse 2004; van Amerongen 2012), which is highly conserved from hydra to humans (Hagemann and others 2014; Hyde and others 2016; Theka and others 2019; Tiwari and others 2015; Wodarz and Nusse 1998). Wnt signaling is classically divided into canonical Wnt/β-catenin signaling pathways and noncanonical Wnt signaling pathways according to the Wnt ligands (Fig.…”

A study of the role of multiple layer‐by‐layer assembled bionic extracellular matrix in promoting wound healing via activation of the Wnt signaling pathway

“…WNT 4, β-catenin, and cyclin D1 represent important marker proteins in the upper, middle, and lower reaches of the Wnt classical pathway (Figure5). β-catenin is considered an important transcription factor in the Wnt pathway, which is transferred from the cell membrane to the nucleus to activate downstream gene of interest 30. Ki67 is a clinical biomarker commonly used to identify cell proliferation, and cyclin D1 can also to a certain extend indicate cell proliferation, thereby determining the correlation between the Wnt signaling pathway and the number of cell proliferation and the degree of wound healing.When the Wnt pathway inhibitor DKk-1 was used to inhibit the activation of the Wnt pathway, the expression levels of important biomarkers such as Wnt4, β-catenin, and cyclin D1 still showed a downward trend even in the culture medium which was supplemented with MLCM (Figure6).…”