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| 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. (en)
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| 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 (en)
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| 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 (en)
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| skos:notation
| - RIV/62157124:16170/13:43872474!RIV14-GA0-16170___
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| http://linked.open...avai/riv/aktivita
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| http://linked.open...avai/riv/aktivity
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| http://linked.open...vai/riv/dodaniDat
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| http://linked.open...aciTvurceVysledku
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| http://linked.open.../riv/druhVysledku
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| http://linked.open...iv/duvernostUdaju
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| http://linked.open...titaPredkladatele
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| http://linked.open...dnocenehoVysledku
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| http://linked.open...ai/riv/idVysledku
| - RIV/62157124:16170/13:43872474
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| http://linked.open...riv/jazykVysledku
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| http://linked.open.../riv/klicovaSlova
| - FMK-inhibitor; micromasses; explant cultures; Caspases (en)
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| http://linked.open.../riv/klicoveSlovo
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| http://linked.open...ontrolniKodProRIV
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| http://linked.open...in/vavai/riv/obor
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| http://linked.open...ichTvurcuVysledku
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| http://linked.open...cetTvurcuVysledku
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| http://linked.open...vavai/riv/projekt
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| http://linked.open...UplatneniVysledku
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| http://linked.open...iv/tvurceVysledku
| - Klepárník, Karel
- Lišková, Marcela
- Matalová, Eva
- Chlastáková, Ivana
- Doubek, Jaroslav
- Kudělová, Judita
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| http://localhost/t...ganizacniJednotka
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