Cell Tracing by a Multicolor Reporter Transgenic Iberian Ribbed Newt Pleurodeles waltl.

Area of Science:

• Regenerative Biology
• Amphibian Research
• Genetics

Background:

• Urodele amphibians possess remarkable regenerative abilities, making them key models in regeneration biology.
• Iberian ribbed newts are increasingly important models due to their reproductive capacity and genetic tractability.
• Cell tracing is crucial for understanding cellular dynamics during organ regeneration.

Purpose of the Study:

• To establish a multicolor reporter system for cell tracing in the Iberian ribbed newt.
• To enable clonal analysis in regenerating tissues of this amphibian model.

Main Methods:

• Generation of transgenic Iberian ribbed newts with the Brainbow3.0 reporter cassette under the CAG promoter.
• Induction of Cre recombinase via electroporation for reporter expression.
• Application of laser ablation for clonal analysis of reporter-positive cells.

Main Results:

• Successful generation of transgenic Brainbow3.0 newts.
• Recombinant reporter expression observed in brain, spinal cord, and muscle tissues.
• Demonstrated ability to trace cells in regenerating tail muscle, midbrain, and spinal cord.
• Enabled clonal analysis of epithelial cells using laser ablation.

Conclusions:

• The Brainbow3.0 reporter system is effectively implemented in Iberian ribbed newts.
• This system facilitates long-term cell tracing and clonal analysis in a key regenerative model.
• The reporter system is expected to be valuable across various research fields in regeneration.