Shenzhen Advanced Institute has developed a new type of high-performance gene-encoded cyclic adenosine phosphate fluorescent probe

Recently, Chu Jun, a researcher at the Biomedical Optics and Molecular Imaging Research Center of the Institute of Biomedical Optics and Health Engineering of the Shenzhen Institute of Advanced Technology of the Chinese Academy of Sciences, published a high-performance genetically encoded fluorescent indicator for in vivo cAMP in Nature Communications Imaging's research paper reports on high-performance gene-encoded cyclic adenosine phosphate (cAMP) fluorescent probes and their applications
.

cAMP is a key second messenger within the cell, integrating signals from multiple G protein-coupled receptors (GPCRs) and playing an important role
in learning and memory, drug addiction, exercise control, immunity, tumors, metabolism, and more.

To develop a highly sensitive probe suitable for in vivo detection, the researchers inserted a cyclized rearranged green fluorescent protein (cpGFP) into the cAMP-binding domain (mlCNBD) of the
bacterial MlotiK1 channel.

In vitro experiments, G-Flamp1 with and without cAMP has different chromophore environments
.

The researchers applied the G-Flamp1 probe to Drosophila, a model organism
.

To verify the usefulness of the G-Flamp1 probe to detect dynamic changes in cAMP in living animals, the researchers used adeno-associated viruses to co-express green G-Flamp1 probes and red jRGECO1a calcium probes in the motor cortex of mice
.

The researchers expressed the G-Flamp1 probe in the nucleus (NAc) brain region of the deep part of the mouse brain and used optical fibers to record changes
in the cAMP signal in this brain region during the auditory Pavlov conditioned reflex task.

This study developed a cAMP fluorescent probe suitable for in vivo detection, and preliminarily revealed the law of cAMP signal changes of specific neurons in specific behavioral processes of model organisms such as fruit flies and mice, laying a foundation
for further elaborating the regulation and function of cAMP signal.

The research work has been funded by the National Key R&D Program and the National Natural Science Foundation of China, and has been supported
by Peking University, the Institute of Neuroscience of the Chinese Academy of Sciences, the Fifth Affiliated Hospital of Sun Yat-sen University, Kansas State University, and Huazhong University of Science and Technology.

Figure 1.

Figure 2: Changes in cAMP signaling in Drosophila Kenyon cells under different stimuli

Figure 3: Changes in cAMP signaling within mouse cortical neurons during exercise

Figure 4.