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Related Concept Videos

The Citric Acid Cycle: Output01:28

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Activity-dependent citrate dynamics in neurons.

Paul C Rosen1,2, Panhui Fu1, Beatriz Ferrán3,4,5

  • 1Department of Neurobiology, Harvard Medical School, Boston, MA 02115.

Proceedings of the National Academy of Sciences of the United States of America
|October 10, 2025
PubMed
Summary
This summary is machine-generated.

Scientists developed new fluorescent biosensors to measure citrate levels in live neurons. They observed a rapid, transient decrease in citrate upon neuronal activation, crucial for understanding glycolysis control.

Keywords:
fluorescence lifetimegenetically encoded fluorescent biosensorglycolytic regulationmitochondrial calcium uniporter

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Area of Science:

  • Neuroscience
  • Metabolic Biochemistry
  • Cellular Physiology

Background:

  • Glycolytic enzymes rapidly adapt cellular energy production in response to metabolic cues.
  • Measuring key glycolytic metabolites, like citrate, with spatiotemporal precision in live cells remains a significant challenge.
  • Neuronal depolarization activates glycolysis, highlighting the need to understand metabolite dynamics during neural activity.

Purpose of the Study:

  • To engineer advanced fluorescent biosensors for precise, real-time measurement of intracellular citrate levels.
  • To investigate the dynamic changes in cytosolic free citrate concentration in neurons during depolarization.
  • To elucidate the role of mitochondrial calcium transport in regulating citrate levels during neuronal activation.

Main Methods:

  • Development of novel quantitative fluorescent biosensors for citrate, optimized for affinity, pH, Mg2+, and temperature.
  • Utilized two-photon fluorescence lifetime imaging microscopy for high-resolution live-cell imaging.
  • Employing acute mouse brain slices to study neuronal responses in a physiologically relevant context.

Main Results:

  • Engineered biosensors successfully quantified citrate dynamics in live neurons.
  • Observed a rapid two-to-threefold decrease in cytosolic free citrate within seconds of neuronal activation.
  • Citrate levels returned to baseline over several minutes, with dependence on the mitochondrial calcium uniporter.

Conclusions:

  • The study presents a breakthrough in live-cell metabolite measurement, enabling real-time monitoring of glycolysis regulators.
  • Demonstrated a rapid, transient decrease in neuronal citrate upon activation, linked to mitochondrial calcium influx.
  • These findings provide critical insights into the fast, dynamic control of glycolysis in neurons.