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Biochemical Assays to Detect Activation of Small GTPases Rho, Rac, and Cdc42 during Morphogenesis

Mark L Berns1, Raymond Habas2

  • 1Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA, USA. mark.berns@temple.edu.

Methods in Molecular Biology (Clifton, N.J.)
|February 11, 2022
PubMed

Insights

This study details biochemical assays to detect Wnt/Frizzled (Fz) pathway activation of Rho, Rac, and Cdc42 GTPases. These assays are crucial for understanding cell polarity and migration in vertebrate development.

Area of Science:

  • Cell Biology
  • Developmental Biology
  • Biochemistry

Background:

  • Wnt/Frizzled (Fz) signaling is a critical pathway regulating diverse biological processes.
  • Activation of small GTPases Rho, Rac, and Cdc42 by Wnt/Fz signaling is a key mechanism for controlling cell polarity and migration.
  • These processes are particularly important during vertebrate gastrulation.

Purpose of the Study:

  • To describe reliable biochemical assays for detecting Wnt/Fz-mediated activation of Rho, Rac, and Cdc42.
  • To provide methods applicable to both mammalian cell cultures and Xenopus embryo explants.

Main Methods:

  • Biochemical assays designed to measure GTPase activity.
  • Application of these assays in mammalian cell lines.
  • Utilizing Xenopus embryo explants to validate assay performance in a developmental context.

Main Results:

  • Successful implementation of biochemical assays to detect Wnt/Fz-induced activation of Rho, Rac, and Cdc42.
  • Demonstration of assay utility in both in vitro mammalian cell systems and in vivo Xenopus models.
  • Quantification of GTPase activation in response to Wnt/Fz pathway stimulation.

Conclusions:

  • The described biochemical assays provide a robust method for studying Wnt/Fz signaling.
  • These assays are valuable tools for investigating the roles of Rho, Rac, and Cdc42 in cell polarity and migration during development.
  • The methods facilitate research into both normal developmental processes and pathological conditions involving Wnt/Fz signaling.
Keywords:
Cdc42GastrulationNoncanonical Wnt signalingRacRhoXenopus

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