A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP

Lobas, Mark A.; Tao, Rongkun; Nagai, Jun; Kronschläger, Mira Therese; Borden, Philip; Marvin, Jonathan S.; Looger, Loren L.; Khakh, Baljit S.

doi:10.1038/s41467-019-08441-5

Cited by 234 publications

(147 citation statements)

References 57 publications

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“…We initially measured ATP concentration in E. coli using a ratiometric fluorescent ATP sensor, mRuby-iATPSnFR1.0 40 . Gentamicin treatment increased the 488/561 nm fluorescence ratio by 50% within 2 hours of treatment ( Fig 5A).…”

Section: Atp Dysregulation Precedes Voltage Induced Bactericidal Killmentioning

confidence: 99%

Membrane voltage dysregulation driven by metabolic dysfunction underlies bactericidal activity of aminoglycosides

Bruni

Kralj

2020

Preprint

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Aminoglycosides are broad-spectrum antibiotics whose mechanism of bactericidal activity has been under debate. It is widely accepted, however, that membrane voltage potentiates aminoglycoside activity, which is ascribed to voltage dependent drug uptake. In this paper, we measured the single cell response of Escherichia coli treated with aminoglycosides and discovered that the bactericidal action arises not from the downstream effects of voltage dependent drug uptake, but rather directly from dysregulated membrane potential. In the absence of voltage, aminoglycosides are taken into cells and exert bacteriostatic effects by inhibiting translation. However, cell killing was immediate upon re-polarization. The hyperpolarization arose from altered ATP flux, which induced a reversal of the F1Fo-ATPase to hydrolyze ATP and generated the deleterious voltage. Heterologous expression of an ATPase inhibitor from Salmonella completely eliminated bactericidal activity, while loss of the F-ATPase significantly reduced the electrophysiological response to aminoglycosides. Our data support a model of voltage induced death, which could be resolved in real-time at the single cell level, and separates the mechanisms of aminoglycoside bacteriostasis and bactericide in E. coli.

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Section: Atp Dysregulation Precedes Voltage Induced Bactericidal Killmentioning

confidence: 99%

Membrane voltage dysregulation driven by metabolic dysfunction underlies bactericidal activity of aminoglycosides

Bruni

Kralj

2020

Preprint

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“…In the present study, we employed the FRET-sensor ATeam for detection of intracellular ATP. Since the description of ATeam and its variants in 2009 (Imamura et al 2009), ATP sensors have been constantly adapted and/or newly designed (Nakano et al 2011;Yaginuma et al 2014;Arai et al 2018;Mendelsohn et al 2018;Lobas et al 2019), also for detection of ATP in organelles (Imamura et al 2009;Rueda et al 2015;Suzuki et al 2018). Moreover, a ratiometric sensor for imaging of the ATP-to-ADP ratio (PercevalHR) is available (Tantama et al 2013).…”

Section: Different Activity Patterns Neuronal Atp Levelsmentioning

confidence: 99%

Relation between activity‐induced intracellular sodium transients and ATP dynamics in mouse hippocampal neurons

Gerkau

Lerchundi

Nelson

et al. 2019

The Journal of Physiology

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Key points Employing quantitative Na+‐imaging and Förster resonance energy transfer‐based imaging with ATeam1.03YEMK (ATeam), we studied the relation between activity‐induced Na+ influx and intracellular ATP in CA1 pyramidal neurons of the mouse hippocampus. Calibration of ATeam in situ enabled a quantitative estimate of changes in intracellular ATP concentrations. Different paradigms of stimulation that induced global Na+ influx into the entire neuron resulted in decreases in [ATP] in the range of 0.1–0.6 mm in somata and dendrites, while Na+ influx that was locally restricted to parts of dendrites did not evoke a detectable change in dendritic [ATP]. Our data suggest that global Na+ transients require global cellular activation of the Na+/K+‐ATPase resulting in a consumption of ATP that transiently overrides its production. For recovery from locally restricted Na+ influx, ATP production as well as fast intracellular diffusion of ATP and Na+ might prevent a local drop in [ATP]. Abstract Excitatory neuronal activity results in the influx of Na+ through voltage‐ and ligand‐gated channels. Recovery from accompanying increases in intracellular Na+ concentrations ([Na+]i) is mainly mediated by the Na+/K+‐ATPase (NKA) and is one of the major energy‐consuming processes in the brain. Here, we analysed the relation between different patterns of activity‐induced [Na+]i signalling and ATP in mouse hippocampal CA1 pyramidal neurons by Na+ imaging with sodium‐binding benzofurane isophthalate (SBFI) and employing the genetically encoded nanosensor ATeam1.03YEMK (ATeam). In situ calibrations demonstrated a sigmoidal dependence of the ATeam Förster resonance energy transfer ratio on the intracellular ATP concentration ([ATP]i) with an apparent KD of 2.6 mm, indicating its suitability for [ATP]i measurement. Induction of recurrent network activity resulted in global [Na+]i oscillations with amplitudes of ∼10 mm, encompassing somata and dendrites. These were accompanied by a steady decline in [ATP]i by 0.3–0.4 mm in both compartments. Global [Na+]i transients, induced by afferent fibre stimulation or bath application of glutamate, caused delayed, transient decreases in [ATP]i as well. Brief focal glutamate application that evoked transient local Na+ influx into a dendrite, however, did not result in a measurable reduction in [ATP]i. Our results suggest that ATP consumption by the NKA following global [Na+]i transients temporarily overrides its availability, causing a decrease in [ATP]i. Locally restricted Na+ transients, however, do not result in detectable changes in local [ATP]i, suggesting that ATP production, together with rapid intracellular diffusion of both ATP and Na+ from and to unstimulated neighbouring regions, counteracts a local energy shortage under these conditions.

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“…In this approach, registering neural activity signals to a common anatomical atlas allows signals from anatomically defined regions to be compared across animals 4 . These tools, combined with the recent development of optical sensors for metabolic flux and red-shifted calcium indicators [14][15][16][17] , allow for the simultaneous imaging of both metabolism and neural activity across the entire brain of the fly. Here, we utilize these sensors to determine the spatiotemporal relationship between metabolic flux and neural activity.…”

mentioning

confidence: 99%

Causal coupling between neural activity, metabolism, and behavior across theDrosophilabrain

Mann

Deny

Ganguli

et al. 2020

Preprint

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Coordinated activity across networks of neurons is a hallmark of both resting and active behavioral states in many species, including worms, flies, fish, mice and humans 1-5 . These global patterns alter energy metabolism in the brain over seconds to hours, making oxygen consumption and glucose uptake widely used proxies of neural activity 6,7 . However, whether changes in neural activity are causally related to changes in metabolic flux in intact circuits on the sub-second timescales associated with behavior, is unknown. Moreover, it is unclear whether transitions between rest and action are associated with spatiotemporally structured changes in neuronal energy metabolism. Here, we combine two-photon microscopy of the entire fruit fly brain with sensors that allow simultaneous measurements of neural activity and metabolic flux, across both resting and active behavioral states. We demonstrate that neural activity drives changes in metabolic flux, creating a tight coupling between these signals that can be measured across large-scale brain networks. Further, these studies reveal that the initiation of even minimal behavioral movements causes large-scale changes in the pattern of neural activity and energy metabolism, revealing unexpected structure in the functional architecture of the brain. The relationship between neural activity and energy metabolism is likely evolutionarily ancient. Thus, these studies provide a critical foundation for using metabolic proxies to capture changes in neural activity and reveal that even minimal behavioral movements are associated with changes in large-scale brain network activity.

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A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP

Cited by 234 publications

References 57 publications

Membrane voltage dysregulation driven by metabolic dysfunction underlies bactericidal activity of aminoglycosides

Membrane voltage dysregulation driven by metabolic dysfunction underlies bactericidal activity of aminoglycosides

Relation between activity‐induced intracellular sodium transients and ATP dynamics in mouse hippocampal neurons

Causal coupling between neural activity, metabolism, and behavior across theDrosophilabrain

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