Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide.

Area of Science:

• Cell Biology
• Biochemistry
• Biophysics

Background:

• Reactive oxygen species (ROS) regulate cellular functions; their overproduction is linked to pathologies.
• Existing genetically encoded ROS sensors emit green light, limiting multiparametric studies due to spectral overlap.
• Developing sensors with different emission spectra is crucial for advanced in vivo ROS detection.

Purpose of the Study:

• To introduce the first genetically encoded red fluorescent sensor for hydrogen peroxide (H2O2) detection.
• To enable multiparametric analysis of ROS production in living systems.
• To demonstrate the utility of the new sensor in cellular and subcellular contexts.

Main Methods:

• Development of a novel red fluorescent protein-based sensor, HyPerRed, for H2O2.
• Validation of HyPerRed's performance against existing green fluorescent sensors.
• Application of HyPerRed to detect H2O2 in the cytoplasm of cultured cells.
• Utilizing HyPerRed to visualize H2O2 dynamics in the mitochondrial matrix.

Main Results:

• HyPerRed exhibits performance comparable to established green fluorescent H2O2 sensors.
• The sensor successfully traced low H2O2 concentrations in cytoplasm upon growth factor stimulation.
• HyPerRed detected localized and transient H2O2 production in mitochondria after inhibiting ER Ca2+ uptake.

Conclusions:

• HyPerRed is a valuable new tool for in vivo detection of H2O2, expanding the capabilities of ROS research.
• The red emission of HyPerRed facilitates multiparametric ROS studies and simultaneous monitoring of other cellular processes.
• This sensor opens new avenues for investigating ROS involvement in various physiological and pathological conditions.