Real-time in vivo imaging of extracellular ATP in the brain with a hybrid-type fluorescent sensor.

Adenosine Triphosphate / analysis Animals Brain / physiology Functional Neuroimaging / methods Male Mice / metabolism Mice, Inbred C57BL Optical Imaging / instrumentation

ATP ATPOS biochemistry chemical biology extracellular signaling molecule fluorescent sensor in vivo imaging mouse neuroscience

Journal

eLife

ISSN: 2050-084X

Titre abrégé: Elife

Pays: England

ID NLM: 101579614

Informations de publication

Date de publication:
10 07 2020

Historique:

received: 03 04 2020

accepted: 09 07 2020

pubmed: 11 7 2020

medline: 24 2 2021

entrez: 11 7 2020

Statut: epublish

Résumé

Adenosine 5' triphosphate (ATP) is a ubiquitous extracellular signaling messenger. Here, we describe a method for in-vivo imaging of extracellular ATP with high spatiotemporal resolution. We prepared a comprehensive set of cysteine-substitution mutants of ATP-binding protein, Biologists often refer to a small molecule called adenosine triphosphate – or ATP for short – as ‘the currency of life’. This molecule carries energy all through the body, and most cells and proteins require ATP to perform their various roles. Nerve cells (also known as neurons) in the brain release ATP when activated, and use this molecule to send signals to other active neurons or other cells in the brain. But ATP can also signal danger in the brain. A molecule derived from ATP is involved in transmitting the pain signals of migraines and severe headaches; and ATP levels can become imbalanced after strokes, when parts of the brain are deprived of blood. Despite its importance, ATP remains difficult to visualize in the body, and monitoring the molecule in the active brain in real time is challenging. To address this issue, Kitajima et al. designed an optical sensor that could monitor ATP in the healthy brain, and was sensitive enough to detect when and where it was released. First, Kitajima et al. made several potential sensors by attaching various fluorescent tags to different locations on a protein that binds ATP. Next each sensor was tested to determine whether it could bind ATP tightly and get bright upon binding. This is important because previous sensors could not detect ATP release in the brains of living animals. To illustrate the new sensors’ potential, Kitajima et al. used the sensor to image ATP in the brains of live mice. A ‘wave’ of ATP was seen spreading through the brain after neurons were stimulated with a small electric pulse, mimicking a sudden migraine or stroke. The results confirm that this new sensor is suitable for imaging how ATP signals in the brain, and it may help resolve the underlying mechanisms of migraines and strokes. This sensor could also be used to understand other cellular process which rely on ATP to carry out their role.

Autres résumés

Type: plain-language-summary (eng)

Biologists often refer to a small molecule called adenosine triphosphate – or ATP for short – as ‘the currency of life’. This molecule carries energy all through the body, and most cells and proteins require ATP to perform their various roles. Nerve cells (also known as neurons) in the brain release ATP when activated, and use this molecule to send signals to other active neurons or other cells in the brain. But ATP can also signal danger in the brain. A molecule derived from ATP is involved in transmitting the pain signals of migraines and severe headaches; and ATP levels can become imbalanced after strokes, when parts of the brain are deprived of blood. Despite its importance, ATP remains difficult to visualize in the body, and monitoring the molecule in the active brain in real time is challenging. To address this issue, Kitajima et al. designed an optical sensor that could monitor ATP in the healthy brain, and was sensitive enough to detect when and where it was released. First, Kitajima et al. made several potential sensors by attaching various fluorescent tags to different locations on a protein that binds ATP. Next each sensor was tested to determine whether it could bind ATP tightly and get bright upon binding. This is important because previous sensors could not detect ATP release in the brains of living animals. To illustrate the new sensors’ potential, Kitajima et al. used the sensor to image ATP in the brains of live mice. A ‘wave’ of ATP was seen spreading through the brain after neurons were stimulated with a small electric pulse, mimicking a sudden migraine or stroke. The results confirm that this new sensor is suitable for imaging how ATP signals in the brain, and it may help resolve the underlying mechanisms of migraines and strokes. This sensor could also be used to understand other cellular process which rely on ATP to carry out their role.

Identifiants

DOI: 10.7554/eLife.57544 PMID: 32648544 PMC: PMC7398694

pubmed: 32648544

doi: 10.7554/eLife.57544

pii: 57544

pmc: PMC7398694

doi:

pii:

Substances chimiques

Adenosine Triphosphate 8L70Q75FXE

Types de publication

Journal Article Research Support, Non-U.S. Gov't