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@article{1786440,
author = {Killinger, Michael and Veselá, Barbora and Procházková, Markéta and Matalová, Eva and Klepárník, Karel},
article_location = {Heidelberg},
article_number = {20},
doi = {http://dx.doi.org/10.1007/s00216-021-03471-9},
keywords = {Cell death and differentiation; Apoptosis; Bioluminescence detection; Caspase-3/7; Single-cell detection and quantification},
language = {eng},
issn = {1618-2642},
journal = {Analytical and Bioanalytical chemistry},
title = {A single-cell analytical approach to quantify activated caspase-3/7 during osteoblast proliferation, differentiation, and apoptosis},
url = {https://doi.org/10.1007/s00216-021-03471-9},
volume = {413},
year = {2021}
}
TY - JOUR
ID - 1786440
AU - Killinger, Michael - Veselá, Barbora - Procházková, Markéta - Matalová, Eva - Klepárník, Karel
PY - 2021
TI - A single-cell analytical approach to quantify activated caspase-3/7 during osteoblast proliferation, differentiation, and apoptosis
JF - Analytical and Bioanalytical chemistry
VL - 413
IS - 20
SP - 5085-5093
EP - 5085-5093
PB - Springer Verlag GmbH
SN - 16182642
KW - Cell death and differentiation
KW - Apoptosis
KW - Bioluminescence detection
KW - Caspase-3/7
KW - Single-cell detection and quantification
UR - https://doi.org/10.1007/s00216-021-03471-9
N2 - The protein heterogeneity at the single-cell level has been recognized to be vital for an understanding of various life processes during animal development. In addition, the knowledge of accurate quantity of relevant proteins at cellular level is essential for appropriate interpretation of diagnostic and therapeutic results. Some low-copy-number proteins are known to play a crucial role during cell proliferation, differentiation, and also in apoptosis. The fate decision is often based on the concentration of these proteins in the individual cells. This is likely to apply also for caspases, cysteine proteases traditionally associated with cell death via apoptosis but recently being discovered also as important factors in cell proliferation and differentiation. The hypothesis was tested in bone-related cells, where modulation of fate from apoptosis to proliferation/differentiation and vice versa is particularly challenging, e.g., towards anti-osteoporotic treatments and anti-cancer strategies. An ultrasensitive and highly selective method based on bioluminescence photon counting was used to quantify activated caspase-3/7 in order to demonstrate protein-level heterogeneity in individual cells within one population and to associate quantitative measurements with different cell fates (proliferation, differentiation, apoptosis). The results indicate a gradual increase of caspase-3/7 activation from the proliferative status to differentiation (more than three times) and towards apoptosis (more than six times). The findings clearly support one of the putative key mechanisms of non-apoptotic functions of pro-apoptotic caspases based on fine-tuning of their activation levels.
ER -
KILLINGER, Michael, Barbora VESELÁ, Markéta PROCHÁZKOVÁ, Eva MATALOVÁ a Karel KLEPÁRNÍK. A single-cell analytical approach to quantify activated caspase-3/7 during osteoblast proliferation, differentiation, and apoptosis. \textit{Analytical and Bioanalytical chemistry}. Heidelberg: Springer Verlag GmbH, 2021, roč.~413, č.~20, s.~5085-5093. ISSN~1618-2642. Dostupné z: https://dx.doi.org/10.1007/s00216-021-03471-9.
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