Filling a GAP—An Optimized Probe for ER Ca 2+ Imaging In Vivo

Malli, Roland; Eroğlu, Emrah; Waldeck-Weiermair, Markus; Graier, Wolfgang F.

doi:10.1016/j.chembiol.2016.06.003

Cited by 3 publications

(2 citation statements)

References 10 publications

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“…Additional disadvantages of NO• sensitive small fluorophores are their potential cytotoxicity and relatively low specificity which make it difficult to use them in a reliable, analytical and conclusive manner789. Although the effective usage of genetically encoded fluorescent probes requires efficient gene transfer techniques, FP-based genetically encoded sensors have emerged as indispensable tools that have revolutionized our understanding of the inner functioning of cells1011. Before the development of the single FP-based geNOps, a Förster resonance energy transfer (FRET)-based NO• sensor, referred to as NOA-112, was constructed.…”

Section: Introductionmentioning

confidence: 99%

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

Eroğlu

Rost

Bischof

et al. 2017

JoVE

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Nitric Oxide (NO•) is a small radical, which mediates multiple important cellular functions in mammals, bacteria and plants. Despite the existence of a large number of methods for detecting NO• in vivo and in vitro, the real-time monitoring of NO• at the single-cell level is very challenging. The physiological or pathological effects of NO• are determined by the actual concentration and dwell time of this radical. Accordingly, methods that allow the single-cell detection of NO• are highly desirable. Recently, we expanded the pallet of NO• indicators by introducing single fluorescent protein-based genetically encoded nitric oxide (NO•) probes (geNOps) that directly respond to cellular NO• fluctuations and, hence, addresses this need. Here we demonstrate the usage of geNOps to assess intracellular NO• signals in response to two different chemical NO•-liberating molecules. Our results also confirm that freshly prepared 3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine (NOC-7) has a much higher potential to evoke change in intracellular NO• levels as compared with the inorganic NO• donor sodium nitroprusside (SNP). Furthermore, dual-color live-cell imaging using the green geNOps (G-geNOp) and the chemical Ca2+ indicator fura-2 was performed to visualize the tight regulation of Ca2+-dependent NO• formation in single endothelial cells. These representative experiments demonstrate that geNOps are suitable tools to investigate the real-time generation and degradation of single-cell NO• signals in diverse experimental setups.

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Section: Introductionmentioning

confidence: 99%

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

Eroğlu

Rost

Bischof

et al. 2017

JoVE

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“…Additional disadvantages of NO• sensitive small fluorophores are their potential cytotoxicity and relatively low specificity which make it difficult to use them in a reliable, analytical and conclusive manner 7,8,9 . Although the effective usage of genetically encoded fluorescent probes requires efficient gene transfer techniques, FP-based genetically encoded sensors have emerged as indispensable tools that have revolutionized our understanding of the inner functioning of cells 10,11 . Before the development of the single FP-based geNOps, a Förster resonance energy transfer (FRET)-based NO• sensor, referred to as NOA-1 12 , was constructed.…”

Section: Introductionmentioning

confidence: 99%

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

Eroğlu

Rost

Bischof

et al. 2017

JoVE

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A HBDI‐Based Fluorescent Probe for Labeling Endoplasmic Reticulum in Living Cells

Ruan

Wang

et al. 2022

Chemistry An Asian Journal

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The endoplasmic reticulum (ER) is an important organelle in eukaryotic cells and is closely involved in the synthesis and processing of proteins, as well as the storage, regulation, and release of calcium. A series of signaling pathways within the ER play a crucial part in the pathogenesis of various diseases, including cancer. Thus, it is necessary to design ER-targeting probes to monitor these signaling pathways. Additionally, precision medicine also requires new ER-targeting group to facilitate the delivery of drug cargoes to the ER. However, only a limited number of ER-targeting groups have been used for the design of fluorescent probes for ER imaging in living cells, as well as the development of ER-targeted drug delivery systems. Herein, a new ER-targeting fluorescent probe (BDI-ER) was designed and prepared. BDI-ER contains the hydrophilic fluorophore, derived from the core structure of GFP, and two hydrophobic octadecane chains. The amphipathic nature of BDI-ER facilitates localization in the ER. Live cell imaging demonstrated selective localization of BDI-ER towards ER compared to other organelles. Additionally, co-localization imaging in various cell lines indicate that BDI-ER is effective at targeting the ER.

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Filling a GAP—An Optimized Probe for ER Ca 2+ Imaging In Vivo

Cited by 3 publications

References 10 publications

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

A HBDI‐Based Fluorescent Probe for Labeling Endoplasmic Reticulum in Living Cells

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Filling a GAP—An Optimized Probe for ER Ca 2+ Imaging In Vivo

Cited by 3 publications

References 10 publications

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells

A HBDI‐Based Fluorescent Probe for Labeling Endoplasmic Reticulum in Living Cells

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Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells