In Vivo Fluorescent Adenosine 5′-Triphosphate (ATP) Imaging of <i>Drosophila melanogaster</i> and <i>Caenorhabditis elegans</i> by Using a Genetically Encoded Fluorescent ATP Biosensor Optimized for Low Temperatures

Tsuyama, Taiichi; Kishikawa, Jun-ichi; Han, Yong-Woon; Harada, Yoshie; Tsubouchi, Asako; Noji, Hiroyuki; Kakizuka, Akira; Yokoyama, Ken; Uemura, Tadashi; Imamura, Hiromi

doi:10.1021/ac4015325

Cited by 101 publications

(101 citation statements)

References 55 publications

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“…To do this, we obtained ATP FRET sensors (ATeams) (10) that are specifically sensitive to ATP (with no response to ADP, other nucleotide triphosphates, NADH, or dATP) (33,34) and are resistant to physiologic changes in pH. In HeLa cells, expression of AT1.03 YEMK (K d ϭ 1.2 mM) gave a robust FRET signal that rapidly decreased when aerobic respiration and glycolysis were inhibited with potassium cyanide (KCN, 1 mM) and 2-deoxyglucose (2DG, 10 mM), respectively (Fig.…”

Section: Aerobic and Glycolytic Atp Requirements Of The Synapticmentioning

confidence: 99%

The Role of Mitochondrially Derived ATP in Synaptic Vesicle Recycling

Pathak

Shields

Mendelsohn

et al. 2015

Journal of Biological Chemistry

259

214

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Background:The ATP requirements of synaptic vesicle release are poorly understood. Results: Mitochondrially derived ATP supports the function of boutons with and without mitochondria. Respiratory dysfunction selectively blocks the reinternalization of synaptic vesicles. Conclusion: ATP diffuses rapidly in axons to support synaptic vesicle recycling. Mitochondrial dysfunction decreases synaptic energy and impairs function. Significance: Understanding energy requirements will help determine how energy failure contributes to neurodegeneration.

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Section: Aerobic and Glycolytic Atp Requirements Of The Synapticmentioning

confidence: 99%

The Role of Mitochondrially Derived ATP in Synaptic Vesicle Recycling

Pathak

Shields

Mendelsohn

et al. 2015

Journal of Biological Chemistry

259

214

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“…After several trials, we decided that 0.1-0.2 pmol of protein injection per oocyte is suitable for the Xenopus ATP imaging system. In this study, we used modifi ed ATeam, which is optimized for the lower temperature of most model animals (20-25 °C) (Tsuyama et al 2013 ). Under these experimental considerations, our strategy worked well, and either 0.1 or 0.2 pmol of ATeam protein also produced (Fig.…”

Section: Injected Ateam Protein Work In Xenopus Oocytesmentioning

confidence: 97%

“…[For the details of ATeam, please refer the original publication (Imamura et al 2009 ).] The number of reports has increased lately about successful ATP observations using ATeam in worms (Kishikawa et al 2012 ), in the hepatitis C virus-replicating cells (Ando et al 2012 ), in plant cells (Hatsugai et al 2012 ), and in fl ies and worms (Tsuyama et al 2013 ) since the fi rst report in HeLa cells (Imamura et al 2009 ).…”

Section: Introductionmentioning

confidence: 99%

ATP Imaging in Xenopus laevis Oocytes

Ijiri

Kishikawa

Imamura

et al. 2014

Sexual Reproduction in Animals and Plants

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Adenosine 5′-triphosphate (ATP) is a major energy currency for various chemical reactions in living cells. In spite of its signifi cant roles, the local ATP timecourse in Xenopus oocyte/egg has not been studied fully yet. Therefore, our goal is to understand the molecular mechanisms of maturation and fertilization by analyzing ATP in oocytes and eggs. A fl uorescence resonance energy transfer (FRET)-based ATP indicator, named ATeam, has been developed and enabled ATP imaging. We are applying this latest experimental technique to observe local ATP in Xenopus oocytes and eggs during maturation and fertilization. First, we used full-grown oocytes to set up the experimental conditions. To observe ATeam fl uorescence in Xenopus oocytes, the translucent oocytes were prepared after injecting ATeam protein into the vegetal hemisphere. Then they were observed under fl uorescence microscopy, and FRET was measured by analyzing their images using software. ATeam displayed strong FRET with low background. More importantly, FRET signal of ATeam consistently increased in response to the addition of ATP, suggesting that ATeam works in the translucent oocytes. Using this Xenopus ATP imaging system, ATP distribution is investigated not only in oocytes during maturation but also in eggs during fertilization.

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“…Genetically encoded molecular tools are also shedding light on molecular processes within the context of intact animals (Akerboom et al, 2009;Dagliyan et al, 2013;Kittelmann et al, 2013;Lissandron et al, 2007;Tsuyama et al, 2013). However, outside of small or optically clear specimens, the ability to penetrate beyond superficial tissue layers to investigate processes in vital tissues remains a major barrier to whole-animal studies.…”

Section: Perspectivesmentioning

confidence: 99%

Genetically encoded molecular probes to visualize and perturb signaling dynamics in living biological systems

Sample

Mehta

Zhang

2014

Journal of Cell Science

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In this Commentary, we discuss two sets of genetically encoded molecular tools that have significantly enhanced our ability to observe and manipulate complex biochemical processes in their native context and that have been essential in deepening our molecular understanding of how intracellular signaling networks function. In particular, genetically encoded biosensors are widely used to directly visualize signaling events in living cells, and we highlight several examples of basic biosensor designs that have enabled researchers to capture the spatial and temporal dynamics of numerous signaling molecules, including second messengers and signaling enzymes, with remarkable detail. Similarly, we discuss a number of genetically encoded biochemical perturbation techniques that are being used to manipulate the activity of various signaling molecules with far greater spatial and temporal selectivity than can be achieved using standard pharmacological or genetic techniques, focusing specifically on examples of chemically driven and light-inducible perturbation strategies. We then describe recent efforts to combine these diverse and powerful molecular tools into a unified platform that can be used to elucidate the molecular details of biological processes that may potentially extend well beyond the realm of signal transduction.

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In Vivo Fluorescent Adenosine 5′-Triphosphate (ATP) Imaging of Drosophila melanogaster and Caenorhabditis elegans by Using a Genetically Encoded Fluorescent ATP Biosensor Optimized for Low Temperatures

Cited by 101 publications

References 55 publications

The Role of Mitochondrially Derived ATP in Synaptic Vesicle Recycling

The Role of Mitochondrially Derived ATP in Synaptic Vesicle Recycling

ATP Imaging in Xenopus laevis Oocytes

Genetically encoded molecular probes to visualize and perturb signaling dynamics in living biological systems

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