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Purine molecules such as adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine (Ado) are widely present inside and outside the cell
.
Intracellular purine molecules are mainly responsible for regulating cell energy metabolism and other processes; while extracellular purine molecules are used as signaling molecules (called "purine transmitters"), which regulate breathing and taste by acting on their corresponding receptors.
On December 22, 2021, Peking University’s Li Yulong Laboratory published a research paper titled A sensitive GRAB sensor for detecting extracellular ATP in vitro and in vivo in Neuron magazine, reporting on the new gene-encoded ATP probe GRABATP1.
0 Development and application in vitro and in live animals
In this work, the Li Yulong laboratory used its previously designed GRAB probe strategy (GPCR Activation-Based sensor) to develop the ATP probe GRABATP1.
0 based on the human ATP receptor P2Y1 and the cyclically rearranged green fluorescent protein cpEGFP.
(Referred to as ATP1.
0)
.
In HEK293T cells, primary neurons and astrocytes cultured in vitro, the ATP1.
Can the ATP1.
0 probe be used to detect endogenously released ATP? The author started with primary cultured hippocampal cells and found that ATP1.
0 can detect ATP release caused by mechanical stimulation and low osmotic pressure stimulation.
Pharmacological experiments and mutant probe experiments further verified the specificity of ATP1.
0 detection signal
.
Interestingly, when no additional stimulation is given, ATP1.
Can the ATP1.
0 probe be used in live animals? Past studies have found that when cells are damaged, the intracellular millimolar level of ATP is released outside the cell, which is sensed by the surrounding glial cells as a "danger signal", thereby activating microglia to release chemokines, etc.
, and produce immunity Response
.
The purine receptors expressed on glial cells play an important role in the process of microglial cell activation, migration and secretion of signal factors
When the brain is in a disease state, what kind of changes will the release of ATP show? As mentioned above, purinergic signals play an important role in immunity
.
In order to detect the changes in the ATP signal in the brain during the immune response, the author triggered a systemic immune response in mice by intraperitoneal injection of lipopolysaccharide (LPS), and at the same time expressed ATP1.
In addition to the development of a highly sensitive ATP1.
0 probe, the author also developed an ATP probe ATP1.
0-L with faster reaction kinetics and lower affinity
.
The affinity (EC50) of ATP1.
In summary, in this work, the author developed a new genetically-encoded ATP fluorescent probe, which achieved high spatio-temporal resolution recording of extracellular ATP
.
Prior to this, Li Yulong's research team also developed another GRAB fluorescent probe for the purine transmitter adenosine in 2020, and helped the Xu Min team at the Center for Excellence in Brain Science and Intelligent Technology of the Chinese Academy of Sciences in the study of sleep regulation
Peking University School of Life Sciences postdoctoral Wu Zhaofa is the first author of this article, Professor Li Yulong and Dr.
Wu Zhaofa are the co-corresponding authors, undergraduate graduate He Kaikai, PhD students Pan Sunlei, Li Bohan, Deng Fei and Wang Huan have made important contributions to the article
.
This work was cooperated by the Jing Miao team (Chen Yue, Xi Fengxue) of the Beijing Brain Science and Brain-like Research Center and the Du Jiulin team (Li Hongyu, Liu Tingting) of the Center for Excellence in Brain Science and Intelligent Technology Innovation of the Chinese Academy of Sciences.
The State Key Laboratory of Membrane Biology, Peking University-Tsinghua Life Sciences Joint Center, National Natural Science Foundation of China, Beijing Municipal Commission of Science and Technology, Feng Foundation and Boehringer Ingelheim Postdoctoral Scholarship and other institutions and funds are strongly supported
.
Original link: https://doi.
org/10.
1016/j.
neuron.
2021.
11.
027
For more details about the work of Li Yulong’s laboratory, please visit: http://yulonglilab.
org/; in addition, Li Yulong’s laboratory is looking for associate researchers, post-doctoral fellows and research assistants with different disciplines and backgrounds.
The treatment is favorable.
Young people interested in brain science are welcome to join.
.
