14 August 2020 |       Share
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A mouse model of hepatic failure was generated by inducing liver apoptosis in
repTOP™ mice with a single intraperitoneal injection of D-GalN+LPS (as shown by immunoblot and caspase activity assay on liver homogenates). Six hours after treatment, mice were given intraperitoneal injections with three different doses of VivoGlo™ Caspase-3/7 Substrate (Promega, Z-DEVD-Aminoluciferin), that is a luciferase pro-substrate activated by the apoptotic enzymes caspase-3/7. Mice were then subjected to a time course analysis carried out with a sequence of in vivo imaging sessions (5 minutes each); photon emission from different body areas of the treated mice was quantified by a CCD-camera and normalized over emission from mice treated with vehicle.  Ex vivo analysis was also carried out in liver and other organs dissected from repTOP™ mice euthanized after the in vivo imaging session. A dose-dependent increase in photon emission, and so of apoptotic activity, was detectable in vivo selectively in the hepatic area of treated animals, and this was confirmed by ex vivo analysis on dissected organs.

Biserni A, Martorana F, Roncoroni C, et al. Identification of apoptotic cells in living reporter mice using modified luciferin: a new avenue to study multiple molecular pathways within the same complex organism. Promega Notes 2009.

An increase in photon emission was detectable in the hepatic area with a maximal cleavage of DEVD-peptide by caspase-3/7 between 15 and 30 minutes afer substrate injection. As a control ex vivo analysis was carried out in liver dissected from repTOP™ mice euthanized after the in vivo imaging session from both vehicle and treated mice.

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