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Glutamine Flux Imaging Using Genetically Encoded Sensors
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Genetically encoded sensors for Chloride concentration.

Claudia Lodovichi1, Gian Michele Ratto2, Andrew J Trevelyan3

  • 1Neuroscience Institute-CNR, Depart. Biomedical Sciences, Unipd, Padova, Veneto Institute of Molecular Medicine, Padova Neuroscience Center, Padova, Italy.

Journal of Neuroscience Methods
|December 24, 2021
PubMed
Summary

Understanding brain function relies on tracking chloride levels in neurons. Genetically-encoded anion biosensors (GABs) enable simultaneous Cl- imaging, but pH interference remains a challenge. New sensors like ClopHensor offer solutions for in vivo measurements.

Keywords:
2-photon. Neuronal excitability. Chloride imaging. Fluorescent proteinsFunctional optical imaging

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Area of Science:

  • Neuroscience
  • Cellular Biology
  • Biochemistry

Background:

  • Intracellular chloride concentration ([Cl-]i) is crucial for neuronal network function, particularly synaptic inhibition.
  • Accurate measurement of [Cl-]i across multiple neurons and subcellular compartments necessitates advanced imaging techniques.
  • Genetically-encoded anion biosensors (GABs) offer cell-type specificity and subcellular resolution for chloride imaging.

Purpose of the Study:

  • To review the historical development and applications of genetically-encoded anion biosensors (GABs) for chloride imaging.
  • To highlight the limitations of current GABs, specifically their sensitivity to both pH and chloride.
  • To introduce ClopHensor as a potential solution for simultaneous in vivo measurement of both ions and discuss future improvements.

Main Methods:

  • Review of historical development and applications of genetically-encoded anion biosensors.
  • Discussion of challenges in disambiguating chloride and pH signals.
  • Introduction and discussion of ClopHensor for in vivo measurements.

Main Results:

  • Genetically-encoded anion biosensors have advanced chloride imaging capabilities in neuroscience.
  • A significant limitation of existing biosensors is their cross-sensitivity to pH.
  • ClopHensor has been developed to address the challenge of simultaneous in vivo pH and chloride measurements.

Conclusions:

  • Genetically-encoded anion biosensors are vital tools for understanding neuronal chloride regulation and brain function.
  • Overcoming pH sensitivity is critical for accurate chloride measurements using biosensors.
  • Future improvements in biosensor technology, such as ClopHensor, promise to enhance our understanding of chloride's role in the brain.