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Visualizing cytoplasmic ATP in C. elegans larvae using PercevalHR.

Aastha Garde1,2, David R Sherwood1

  • 1Department of Biology, Duke University, Box 90338, Durham, NC 27708, USA.

STAR Protocols
|June 6, 2022
PubMed
Summary
This summary is machine-generated.

This study details a method to measure cellular energy (ATP levels) in living animals using the PercevalHR biosensor. This protocol quantifies the ATP:ADP ratio in *C. elegans* during key developmental events.

Keywords:
CancerCell BiologyDevelopmental biologyMetabolismMicroscopyModel OrganismsMolecular/Chemical Probes

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Area of Science:

  • Cellular Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Measuring intracellular adenosine triphosphate (ATP) levels is crucial for understanding cellular energy metabolism in living organisms.
  • Challenges in quantifying ATP include tissue complexity and limitations of existing biosensors, particularly in whole-animal studies.
  • The ATP:ADP ratio serves as a key indicator of cellular energy status.

Purpose of the Study:

  • To present a protocol for quantifying cytoplasmic ATP levels in living *C. elegans* larvae.
  • To adapt and apply the PercevalHR fluorescent biosensor for measuring the ATP:ADP ratio during anchor cell invasion.
  • To provide a method adaptable for analyzing ATP:ADP ratios in various *C. elegans* cell types.

Main Methods:

  • Utilized the PercevalHR fluorescent biosensor, which reports the cellular ATP:ADP ratio.
  • Applied the protocol to *C. elegans* larvae, specifically during the process of anchor cell invasion.
  • Focused on quantifying ATP levels within the cytosol of living cells.

Main Results:

  • Successfully demonstrated the quantification of ATP:ADP ratios in *C. elegans* larvae using PercevalHR.
  • Established a viable protocol for in vivo measurement of cellular energy status during a specific developmental process.
  • Validated the utility of PercevalHR for assessing cytoplasmic energy dynamics.

Conclusions:

  • The described protocol enables effective measurement of cellular ATP levels in *C. elegans*.
  • PercevalHR is a valuable tool for studying cellular energetics in vivo, adaptable to different cell types.
  • This method advances the understanding of energy support for cellular activities in complex biological systems.