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Validation and application of caged Z-DEVD-aminoluciferin bioluminescence for assessment of apoptosis of wild type and TLR2-deficient mice after ischemic stroke

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Title: Validation and application of caged Z-DEVD-aminoluciferin bioluminescence for assessment of apoptosis of wild type and TLR2-deficient mice after ischemic stroke
Authors: Josić Dominović, P., Dobrivojević Radmilović, M., Srakočić, S., Mišerić, I., Škokić, S., Gajović, Srećko
Source: Journal of Photochemistry and Photobiology B: Biology. 253:112871
Publisher Information: Elsevier BV, 2023.
Publication Year: 2023
Subject Terms: Mice, Toll-Like Receptor 2 / genetics, Caspase 3, Caspase 3 / metabolism, Toll-Like Receptor 2 / metabolism, Animals, Apoptosis, Firefly Luciferin / analogs & derivatives, Firefly Luciferin, Oligopeptides, Toll-Like Receptor 2, Ischemic Stroke
Description: Programmed cell death or apoptosis is a critically important mechanism of tissue remodeling and regulates conditions such as cancer, neurodegeneration or stroke. The aim of this research article was to assess the caged Z-DEVD-aminoluciferin substrate for in vivo monitoring of apoptosis after ischemic stroke in TLR2-deficient mice and their TLR2-expressing counterparts. Postischemic inflammation is a significant contributor to ischemic injury development and apoptosis, and it is modified by the TLR2 receptor. Caged Z-DEVD-aminoluciferin is made available for bioluminescence enzymatic reaction by cleavage with activated caspase-3, and therefore it is assumed to be capable of reporting and measuring apoptosis. Apoptosis was investigated for 28 days after stroke in mice which ubiquitously expressed the firefly luciferase transgene. Middle cerebral artery occlusion was performed to achieve ischemic injury, which was followed with magnetic resonance imaging. The scope of apoptosis was determined by bioluminescence with caged Z-DEVD-aminoluciferin, immunofluorescence with activated caspase-3, flow cytometry with annexin-V and TUNEL assay. The linearity of Z-DEVD-aminoluciferin substrate dose effect was shown in the murine brain. Z-DEVD-aminoluciferin was validated as a good tool for monitoring apoptosis following adequate adjustment. By utilizing bioluminescence of Z-DEVD-aminoluciferin after ischemic stroke it was shown that TLR2-deficient mice had lower post-stroke apoptosis than TLR2-expressing wild type mice. In conclusion, Z-DEVD-aminoluciferin could be a valuable tool for apoptosis measurement in living mice.
Document Type: Article
File Description: application/pdf
Language: English
ISSN: 1011-1344
DOI: 10.1016/j.jphotobiol.2024.112871
DOI: 10.2139/ssrn.4633393
Access URL: https://pubmed.ncbi.nlm.nih.gov/38402658
https://urn.nsk.hr/urn:nbn:hr:105:271637
https://doi.org/10.1016/j.jphotobiol.2024.112871
Rights: CC BY
Accession Number: edsair.doi.dedup.....ebc8bc6cd5d7553a044cd3b1e9f9bc3d
Database: OpenAIRE
Description
Abstract:Programmed cell death or apoptosis is a critically important mechanism of tissue remodeling and regulates conditions such as cancer, neurodegeneration or stroke. The aim of this research article was to assess the caged Z-DEVD-aminoluciferin substrate for in vivo monitoring of apoptosis after ischemic stroke in TLR2-deficient mice and their TLR2-expressing counterparts. Postischemic inflammation is a significant contributor to ischemic injury development and apoptosis, and it is modified by the TLR2 receptor. Caged Z-DEVD-aminoluciferin is made available for bioluminescence enzymatic reaction by cleavage with activated caspase-3, and therefore it is assumed to be capable of reporting and measuring apoptosis. Apoptosis was investigated for 28 days after stroke in mice which ubiquitously expressed the firefly luciferase transgene. Middle cerebral artery occlusion was performed to achieve ischemic injury, which was followed with magnetic resonance imaging. The scope of apoptosis was determined by bioluminescence with caged Z-DEVD-aminoluciferin, immunofluorescence with activated caspase-3, flow cytometry with annexin-V and TUNEL assay. The linearity of Z-DEVD-aminoluciferin substrate dose effect was shown in the murine brain. Z-DEVD-aminoluciferin was validated as a good tool for monitoring apoptosis following adequate adjustment. By utilizing bioluminescence of Z-DEVD-aminoluciferin after ischemic stroke it was shown that TLR2-deficient mice had lower post-stroke apoptosis than TLR2-expressing wild type mice. In conclusion, Z-DEVD-aminoluciferin could be a valuable tool for apoptosis measurement in living mice.
ISSN:10111344
DOI:10.1016/j.jphotobiol.2024.112871