A fluorescent reporter of caspase activity for live imaging

• DOI: 10.1073/pnas.0806983105
• Publication Date: 2008-09-09T00:55:04Z
• Authors: Pierre-Luc Bardet; Golnar Kolahgar; Anita Mynett; Irene Miguel-Aliaga; James Briscoe; Pascal Meier; Jean-Paul Vincent
• Keywords: Cleavage (geology)

There is a growing interest in the mechanisms that control apoptosis cascade during development and adult life. To investigate regulatory events trigger whole tissues, we have devised genetically encoded caspase sensor can be detected live fixed tissue by standard confocal microscopy. The comprises two fluorophores, mRFP, monomeric red fluorescent protein (mRFP) enhanced green (eGFP), are linked an efficient specific caspase-sensitive site. Upon activation, cleaved eGFP translocates to nucleus, leaving mRFP at membranes. This before other markers of apoptosis, including anti-cleaved 3 immunoreactivity. Moreover, does not perturb normal developmental specific, as cleavage occur Drosophila embryos unable activate apoptotic cascade. Importantly, dying cells recognized embryos, thus opening way for vivo imaging. As expected from high conservation caspases, it also chick embryos. It therefore likely generally useful track spatiotemporal pattern activity variety species.