Dynamics of caspase-3 activation and inhibition in embryonic micromasses evaluated by a photon-counting chemiluminescence approach
Language English Country Germany Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Apoptosis drug effects MeSH
- Mice, Inbred Strains MeSH
- Caspase Inhibitors pharmacology MeSH
- Camptothecin pharmacology MeSH
- Caspase 3 metabolism physiology MeSH
- Cells, Cultured MeSH
- Luminescent Measurements methods MeSH
- Mice MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Caspase Inhibitors MeSH
- Camptothecin MeSH
- Caspase 3 MeSH
Caspases are key enzymatic components of the intracellular apoptotic machinery, and their role in mammalian systems is often studied using fluoromethylketone (FMK) inhibitors. Despite many advantages of such approach, efficiency of the inhibitor and membrane permeability speed are often questioned. This work therefore focuses on an exact evaluation of caspase-3 FMK inhibition dynamics in camptothecin-induced mesenchymal micromasses. Two parameters of caspase-3 FMK inhibitor were investigated: first, the stability of the inhibitory potential in the time course of cultivation and, simultaneously, the dynamics of caspase-3 FMK inhibition after camptothecin-induced apoptosis peak. A photon-counting chemiluminescence approach was applied for quantification of active caspase-3. The sensitivity of the photon-counting method allowed for evaluation of active caspase-3 concentration in femtogram amounts per cell. The inhibitor penetrated the cells within the first minute after its application, and the peak of caspase-3 started to decline to the blank level after 30 min. The inhibitory effect of the FMK inhibitor was unchanged during the entire 48 h of cultivation.
See more in PubMed
Cell. 1998 Aug 7;94(3):325-37 PubMed
Cell. 1998 Aug 7;94(3):339-52 PubMed
J Cell Biochem. 2010 Oct 15;111(3):653-8 PubMed
Nature. 1996 Nov 28;384(6607):368-72 PubMed
Immunity. 1998 Aug;9(2):267-76 PubMed
Cell Death Differ. 2009 Jan;16(1):3-11 PubMed
Biochemistry. 1999 Oct 19;38(42):13906-11 PubMed
Arch Oral Biol. 2010 Aug;55(8):570-5 PubMed
Biophys J. 2005 Oct;89(4):2770-82 PubMed
J Biol Chem. 1985 Nov 25;260(27):14873-8 PubMed
EMBO J. 2011 Aug 23;30(18):3667-83 PubMed
Biol Pharm Bull. 2011;34(2):290-4 PubMed
Curr Biol. 1999 Sep 9;9(17):967-70 PubMed
Toxicol In Vitro. 2011 Oct;25(7):1385-91 PubMed
Trends Cell Biol. 2004 Apr;14(4):184-93 PubMed
Cell Death Differ. 2001 Oct;8(10):985-94 PubMed
Caspase Inhibition Affects the Expression of Autophagy-Related Molecules in Chondrocytes
Osteogenic impact of pro-apoptotic caspase inhibitors in MC3T3-E1 cells
Osteogenic Potential of Caspases Related to Endochondral Ossification
Caspases and osteogenic markers--in vitro screening of inhibition impact