The method of ablating different cells is conducive to the study of cell function or regeneration. Current chemical / genetic coding systems have limited spatial resolution, but optogenetic tools provide unique selectivity to target cells.
Zebrafish (Danio rerio) is a popular vertebrate model organism used to study the molecular mechanisms driving development and disease. The beneficial optical properties of zebrafish allow not only passive observation, but also active manipulation of proteins and cells through light using optogenetic tools. Creative BioMart has been able to build an optical cell ablation system on zebrafish for our customers.
In principle, almost any visible cell in zebrafish embryos can be ablated by laser irradiation. The power output of the laser and the type of optical system used to focus the beam on the battery will determine whether a specific battery can be ablated in practice. The following protocol has been successfully used to excise spinal cord, single floor, muscle, notochord and neural crest cells, as well as single identified neurons in the whole ganglion.
Killer red (KR) is a fluorescent protein, which produces a large dose of reactive oxygen species (ROS) under green light irradiation, leading to apoptosis. KR has been used for ROS induced cell ablation in zebrafish. Transgenic fish lines expressing membrane tethered Kr in heart or kidney were produced. ROS levels can be controlled to cause damage to cell membranes or death of Kr positive cells. In addition, the activation of Kr in the heart is very similar to human heart failure. In order to study neurodegeneration, we made a transgenic zebrafish model expressing Kr in spinal cord neurons to provide single-cell control for cell ablation.
The possibility of controlling the area, intensity and duration of ROS expression makes Kr transgenic model a useful tool, which can be used not only for targeted cell ablation, but also to study the effects of free radicals on different tissues.
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