Dynamics of caspase-3 activation and inhibition in embryonic
micromasses evaluated by a photon-counting chemiluminescence
approach

J 2012

Dynamics of caspase-3 activation and inhibition in embryonic micromasses evaluated by a photon-counting chemiluminescence approach

CHLASTAKOVA, I; Marcela LIŠKOVÁ; Jarmila KUDELOVÁ; Lenka ZDRAŽILOVÁ DUBSKÁ; Karel KLEPÁRNÍK et al.

Základní údaje

Originální název

Dynamics of caspase-3 activation and inhibition in embryonic micromasses evaluated by a photon-counting chemiluminescence approach

Autoři

CHLASTAKOVA, I; Marcela LIŠKOVÁ; Jarmila KUDELOVÁ; Lenka ZDRAŽILOVÁ DUBSKÁ; Karel KLEPÁRNÍK a Eva MATALOVÁ

Vydání

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 2012

Další údaje

Typ výsledku

Článek v odborném periodiku

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

DOI

https://doi.org/10.1007/s11626-012-9542-8

UT WoS

000310427000001

Změněno: 15. 10. 2015 11:53, Ing. Mgr. Věra Pospíšilíková

Anotace

V originále

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.

Citovat

CHLASTAKOVA, I; Marcela LIŠKOVÁ; Jarmila KUDELOVÁ; Lenka ZDRAŽILOVÁ DUBSKÁ; Karel KLEPÁRNÍK a Eva MATALOVÁ. Dynamics of caspase-3 activation and inhibition in embryonic micromasses evaluated by a photon-counting chemiluminescence approach. IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL. 2012, roč. 48, č. 9, s. 545-549. Dostupné z: https://doi.org/10.1007/s11626-012-9542-8.

@article{1202735,
   author = {Chlastakova, I and Lišková, Marcela and Kudelová, Jarmila and Zdražilová Dubská, Lenka and Klepárník, Karel and Matalová, Eva},
   article_number = {9},
   doi = {https://doi.org/10.1007/s11626-012-9542-8},
   journal = {IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL},
   title = {Dynamics of caspase-3 activation and inhibition in embryonic micromasses evaluated by a photon-counting chemiluminescence approach},
   volume = {48},
   year = {2012}
}

TY  - JOUR
ID  - 1202735
AU  - Chlastakova, I - Lišková, Marcela - Kudelová, Jarmila - Zdražilová Dubská, Lenka - Klepárník, Karel - Matalová, Eva
PY  - 2012
TI  - Dynamics of caspase-3 activation and inhibition in embryonic micromasses evaluated by a photon-counting chemiluminescence approach
JF  - IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
VL  - 48
IS  - 9
SP  - 545-549
EP  - 545-549
N2  - 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.
ER  -

CHLASTAKOVA, I; Marcela LIŠKOVÁ; Jarmila KUDELOVÁ; Lenka ZDRAŽILOVÁ DUBSKÁ; Karel KLEPÁRNÍK a Eva MATALOVÁ. Dynamics of caspase-3 activation and inhibition in embryonic micromasses evaluated by a photon-counting chemiluminescence approach. \textit{IN VITRO CELLULAR \&{}amp; DEVELOPMENTAL BIOLOGY-ANIMAL}. 2012, roč.~48, č.~9, s.~545-549. Dostupné z: https://doi.org/10.1007/s11626-012-9542-8.

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