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Genetically Targeted Fluorescent Probes Reveal Dynamic Calcium Responses to Adrenergic Signaling in Multiple Cardiomyocyte Compartments.

08:00 EDT 9th July 2019 | BioPortfolio

Summary of "Genetically Targeted Fluorescent Probes Reveal Dynamic Calcium Responses to Adrenergic Signaling in Multiple Cardiomyocyte Compartments."

Calcium (Ca), an important second messenger, regulates many cellular activities and varies spatiotemporally within the cell. Conventional methods to monitor Ca changes, such as synthetic Ca indicators, are not targetable, while genetically encoded Ca indicators (GECI) can be precisely directed to cellular compartments. GECIs are chimeric proteins composed of calmodulin (or other protein that changes conformation on Ca binding) coupled with two fluorescent proteins that come closer together after an increase in [Ca], and enhance Förster energy transfer (FRET) that allows for ratiometric [Ca] assessment. Here, adult rat ventricular myocytes were transfected with specifically targeted calmodulin-based GECIs and Ca responses to a physiological stimulus, norepinephrine (NE, 10 µM), were observed in a) sarcoplasmic reticulum (SR), b) mitochondria, c) the space between the mitochondria and SR, termed the Mitochondria Associated Membrane space (MAM) and d) cytosol for 10 min after stimulation. In SR and mitochondria, NE increased the [Ca] ratio by 17% and by 8%, respectively. In the MAM the [Ca] ratio decreased by 16%, while in cytosol [Ca] remained unchanged. In conclusion, adrenergic stimulation causes distinct responses in the cardiomyocyte SR, mitochondria and MAM. Additionally, our work provides a toolkit-update for targeted [Ca] measurements in multiple cellular compartments.

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This article was published in the following journal.

Name: The international journal of biochemistry & cell biology
ISSN: 1878-5875
Pages: 105569

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Medical and Biotech [MESH] Definitions

A fluorescent calcium chelating agent which is used to study intracellular calcium in many tissues. The fluorescent and chelating properties of Fura-2 aid in the quantitation of endothelial cell injury, in monitoring ATP-dependent calcium uptake by membrane vesicles, and in the determination of the relationship between cytoplasmic free calcium and oxidase activation in rat neutrophils.

The use of light interaction (scattering, absorption, and fluorescence) with biological tissue to obtain morphologically based information. It includes measuring inherent tissue optical properties such as scattering, absorption, and autofluorescence; or optical properties of exogenous targeted fluorescent molecular probes such as those used in optical MOLECULAR IMAGING, or nontargeted optical CONTRAST AGENTS.

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