Question

1. In Zebrafish what novel insights are stable transgenic lines expressing fluorescent providing?

Answer 1

Zebrafish are an ideal model organism to research cancer.

A. Zebrafish embryos and larvae are optically translucent, which has made imaging multiple processes in development and disease possible. When coupled with fluorescent imaging techniques, zebrafish are fast becoming a model of choice for following tumor formation. This is highlighted by recent studies using fluorescent proteins to image xenograft transplantation, neovascularization, growth responses to drug treatments, and self-renewal.

B. Fluorescent labeled tumors can be generated in zebrafish by multiple methods including chemical mutagenesis, oncogene expression by mosaic or stable transgenesis, or genetic mutations that are predisposing to cancer.

C. Numerous fluorescent protein variants are currently available and can be used in imaging cancer in zebrafish. For example, E2-crimson is a far shifted red fluorescent protein that can be excited with a 633 nm laser and could be used in combination with multiple fluorescent reporters. Most experimental models have either used GFP or red fluorescent proteins to distinguish tumor cell subpopulations because of the extreme spectral differences between these proteins. However, other fluorescent protein combinations can be used. For example, we have had good success in imaging membrane localized blue fluorescence protein (lyn-amCyan) from GFP and monomeric RFP within the same animal using confocal imaging.