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Statements

Subject Item
n2:RIV%2F62157124%3A16170%2F13%3A43872474%21RIV14-GA0-16170___
rdf:type
skos:Concept n15:Vysledek
dcterms:description
Caspases are key molecules of physiological cell death activated in proteolytic cascade and play a fundamental role during embryonic and postnatal development, particularly in apoptotic events. Caspase-3 is a central caspase of apoptosis, therefore, a common target for manipulations in research and development of anti-cancer therapies. Fluoromethylketones (FMK) are widely used for pharmacological inhibition of caspases, nevertheless, several questions related to inhibitor penetrability, stability and inhibition dynamics remain unclear. Mouse limb digitalization represents the most well known example of apoptosis. This work focuses on caspase-3 inhibition by FMK-inhibitor at the stage prior to the interdigital regression (E12). Two different ex vivo/in vitro approaches, explants cultures and mesenchymal micromasses, were exploited to investigate two aspects. The effect of caspase-3 inhibition on limb digitalization was followed using front limb cultures. Inhibition dynamics was evaluated in camptothecine induced limb mesenchymal cells using a novel method of photon counting chemiluminiscence. Caspase-3 inhibition in the limb explants caused a delay in onset of interdigital apoptosis compared to the control group but not a complete block. This supports a hypothesis of compensatory mechanisms as in the case of deficiency of other apoptotic component such as Apaf-1. The inhibitor penetrates the cell during the first five minutes of contact (medium delivery by diffusion). The complete inhibition effect (a blank level of active caspase-3) was observed after 30 min. The FMK-inhibitory effect showed constant stability during 48 h of cultivation (limit for medium exchange). Evaluation of inhibitory dynamics was achieved with accuracy of femtograms of active caspase-3 per cell. Caspases are key molecules of physiological cell death activated in proteolytic cascade and play a fundamental role during embryonic and postnatal development, particularly in apoptotic events. Caspase-3 is a central caspase of apoptosis, therefore, a common target for manipulations in research and development of anti-cancer therapies. Fluoromethylketones (FMK) are widely used for pharmacological inhibition of caspases, nevertheless, several questions related to inhibitor penetrability, stability and inhibition dynamics remain unclear. Mouse limb digitalization represents the most well known example of apoptosis. This work focuses on caspase-3 inhibition by FMK-inhibitor at the stage prior to the interdigital regression (E12). Two different ex vivo/in vitro approaches, explants cultures and mesenchymal micromasses, were exploited to investigate two aspects. The effect of caspase-3 inhibition on limb digitalization was followed using front limb cultures. Inhibition dynamics was evaluated in camptothecine induced limb mesenchymal cells using a novel method of photon counting chemiluminiscence. Caspase-3 inhibition in the limb explants caused a delay in onset of interdigital apoptosis compared to the control group but not a complete block. This supports a hypothesis of compensatory mechanisms as in the case of deficiency of other apoptotic component such as Apaf-1. The inhibitor penetrates the cell during the first five minutes of contact (medium delivery by diffusion). The complete inhibition effect (a blank level of active caspase-3) was observed after 30 min. The FMK-inhibitory effect showed constant stability during 48 h of cultivation (limit for medium exchange). Evaluation of inhibitory dynamics was achieved with accuracy of femtograms of active caspase-3 per cell.
dcterms:title
Dynamics of caspase-3 inhibition in embryonic mouse limbs after developmentally and experimentally induced apoptosis Dynamics of caspase-3 inhibition in embryonic mouse limbs after developmentally and experimentally induced apoptosis
skos:prefLabel
Dynamics of caspase-3 inhibition in embryonic mouse limbs after developmentally and experimentally induced apoptosis Dynamics of caspase-3 inhibition in embryonic mouse limbs after developmentally and experimentally induced apoptosis
skos:notation
RIV/62157124:16170/13:43872474!RIV14-GA0-16170___
n15:predkladatel
n17:orjk%3A16170
n3:aktivita
n12:P
n3:aktivity
P(GAP502/12/1285)
n3:dodaniDat
n18:2014
n3:domaciTvurceVysledku
n13:4002032 n13:9388583 n13:8914427 n13:3362051
n3:druhVysledku
n10:O
n3:duvernostUdaju
n8:S
n3:entitaPredkladatele
n9:predkladatel
n3:idSjednocenehoVysledku
70955
n3:idVysledku
RIV/62157124:16170/13:43872474
n3:jazykVysledku
n14:eng
n3:klicovaSlova
FMK-inhibitor; micromasses; explant cultures; Caspases
n3:klicoveSlovo
n5:FMK-inhibitor n5:Caspases n5:explant%20cultures n5:micromasses
n3:kontrolniKodProRIV
[708564E36B41]
n3:obor
n6:GJ
n3:pocetDomacichTvurcuVysledku
4
n3:pocetTvurcuVysledku
6
n3:projekt
n16:GAP502%2F12%2F1285
n3:rokUplatneniVysledku
n18:2013
n3:tvurceVysledku
Matalová, Eva Chlastáková, Ivana Doubek, Jaroslav Lišková, Marcela Kudělová, Judita Klepárník, Karel
n11:organizacniJednotka
16170