In vivo oxidative stress measurements during early life stages.

Transgene model for in vivo analyses of redox potential.

In vivo oxidative stress measurements during early life stages.
Start date: 1. January 2014
End date: 31. December 2014
Financed by: NFD
The project is lead by: NIFES

Background

The redox potential is important for control of metabolism and is affected by both nutrients and contaminants. The redox potential will be different in different organs, in intra- and extracellular spaces, during proliferation/differentiation/cell death, and in different organelles. Changes in the redox potential also occurs during embryo development and organ development at larval stages. In zebrafish embryo and larvae, the degree of oxidation in tissues and organs can be measured in vivo by utilizing a redox sensitive variant of the green fluorescent protein; roGFP. We plan to establish a stable, transgenic zebrafish roGFP line for in vivo quantitative and qualitative redox potentials during early life stages. This will generate basic knowledge for further studies of how nutrients and contaminant affect the redox potential during development.

Objective

We will characterize the transgenic Tg;RedOX line by performing two different pilot-experiments:

  1. During development, the cells in embryos and larvae undergo proliferation, differentiation and apoptosis. The redox potential of a cell will vary depending on its status; during proliferation the cellular environment is reduced, while differentiation leaves the cell in a more oxidized state. Apoptosis is characterized by an even further increased oxidized state. By measuring the redox potential in different compartments of the intestine and in muscle during development, utilizing the roGFP transgenic line, we aim to characterize the role of the redox potential during the developmental processes of these tissues.
  2. Replacement of marine ingredients with vegetable feed ingredients in aquaculture species, new contaminants are introduced, including pesticides. Pesticides are toxic substances that can cause oxidative stress, and are very often shown to be neurotoxic. We will investigate the degree of ROS formation in the developing CNS, utilizing the Tg;RedOx transgenic zebrafish line in combination with embryos exposed for the pesticides Chlorpyrifos and Endosulphan.

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