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Click on the blue letters to pay attention to our glial cells are the collective names of oligodendrocytes, astrocytes, oligodendrocyte progenitor cells, ependymal cells, microglia and other cells.
In damage or infection of the central nervous system, both microglia and astrocytes undergo morphological, molecular and functional remodeling.
How do the "two brothers" microglia and astrocytes, both of which belong to glial cells, conduct an efficient "talk".
In autoimmune brain diseases such as multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), the activation of astrocytes is regulated by T cells and other peripheral immune cells.
Although the current single-cell sequencing technology is relatively mature, it cannot easily capture the interactions between some cell populations.
On April 23, 2021, the Harvard Medical School’s Brigham and Women’s Hospital Francisco J.
Quintana research team published an article in Science, and developed a viral vector-based single-cell sequencing technology that can effectively realize the interaction between cells.
Recognition of molecular targets.
Researchers have developed RABID-seq, a single-cell sequencing technology that recognizes astrocyte interactions.
They first constructed a glycoprotein-deficient rabies virus (RabDG) capable of expressing fluorescent mRNA barcodes.
Since pseudorabies virus infects cells expressing envelope protein A or envelope protein A receptors, the two types of interacting cells will carry mRNA "barcodes", which can then be single-cell sequencing.
RabDG experimental labeling process.
After injecting RabDG virus into GFAP transgenic mice expressing glycosyl proteins and cell membrane receptors, the virus first infects astrocytes that can express cell membrane receptors, and then these astrocytes Plasma cells are labeled with a "barcode" of the mRNA.In addition, new virus particles produced by astrocytes integrate rabies glycoprotein G into their envelope, thereby gaining the ability to infect neighboring cells.
In this way, the molecular mechanism of the interaction between astrocytes and their neighboring cells can be studied.
The virus was further injected into EAE model mice, and 32880 cells with RabDG QR codes were analyzed by RABID-seq technology, including astrocytes, microglia, monocytes, and T cells.
They found that inflammatory signals activated by microglia and chemokine-related signaling pathways are closely related to highly pro-inflammatory astrocytes.
Sema4D is a cell surface protein of T cells involved in lymphocyte activation, and PlexinB2 is its receptor.
The researchers found that the Sema4D-PlexinB2 signaling pathway was activated during the interaction between EAE model mouse microglia and astrocytes.
In addition, Sema4d was expressed on microglia, while PlexinB2 was expressed on astrocytes , Sema4D and PlexinB2 expressions were up-regulated in EAE model mice.
PlexinB2-positive astrocytes prefer to be close to Sema4d-positive microglia.
Further inhibiting the activities of Sema4D and PlexinB2 by CRISPR-Cas9 technology can effectively alleviate the symptoms of EAE model mice.
The researchers found out the treasure signal Sema4D-PlexinB2 through RABID-seq, and also found another pair of treasure signals EphB3-Ephrin-B3.
Erythropoietin human hepatocyte B (EphB) receptor is a member of the receptor tyrosine kinase family, and it works by binding to its ligand EphfinB.
The expression of EphB3-Ephrin-B3 on glial cells The expression of Efnb3 on microglia in EAE model mice increased, which promoted the inflammatory activation of microglia; the expression of EphB3 on astrocytes also increased.
After interfering RNA technology reduces the expression of Efnb3 on microglia and EphB3 on astrocytes, the inflammation activation state is significantly reduced.
Inhibition of EphB3 receptor can also significantly improve the symptoms of EAE model mice.
In summary, this article has discovered two signaling molecular pathways that "talk" between microglia and astrocytes through the development of a new cell-interacting single-cell sequencing technology RABID-seq: EphB3-Ephrin-B3 and Sema4D-PlexinB2.
[References] 1.
https://doi.
org/10.
1126/science.
abf1230 The pictures in the article are all from the references
In damage or infection of the central nervous system, both microglia and astrocytes undergo morphological, molecular and functional remodeling.
How do the "two brothers" microglia and astrocytes, both of which belong to glial cells, conduct an efficient "talk".
In autoimmune brain diseases such as multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), the activation of astrocytes is regulated by T cells and other peripheral immune cells.
Although the current single-cell sequencing technology is relatively mature, it cannot easily capture the interactions between some cell populations.
On April 23, 2021, the Harvard Medical School’s Brigham and Women’s Hospital Francisco J.
Quintana research team published an article in Science, and developed a viral vector-based single-cell sequencing technology that can effectively realize the interaction between cells.
Recognition of molecular targets.
Researchers have developed RABID-seq, a single-cell sequencing technology that recognizes astrocyte interactions.
They first constructed a glycoprotein-deficient rabies virus (RabDG) capable of expressing fluorescent mRNA barcodes.
Since pseudorabies virus infects cells expressing envelope protein A or envelope protein A receptors, the two types of interacting cells will carry mRNA "barcodes", which can then be single-cell sequencing.
RabDG experimental labeling process.
After injecting RabDG virus into GFAP transgenic mice expressing glycosyl proteins and cell membrane receptors, the virus first infects astrocytes that can express cell membrane receptors, and then these astrocytes Plasma cells are labeled with a "barcode" of the mRNA.In addition, new virus particles produced by astrocytes integrate rabies glycoprotein G into their envelope, thereby gaining the ability to infect neighboring cells.
In this way, the molecular mechanism of the interaction between astrocytes and their neighboring cells can be studied.
The virus was further injected into EAE model mice, and 32880 cells with RabDG QR codes were analyzed by RABID-seq technology, including astrocytes, microglia, monocytes, and T cells.
They found that inflammatory signals activated by microglia and chemokine-related signaling pathways are closely related to highly pro-inflammatory astrocytes.
Sema4D is a cell surface protein of T cells involved in lymphocyte activation, and PlexinB2 is its receptor.
The researchers found that the Sema4D-PlexinB2 signaling pathway was activated during the interaction between EAE model mouse microglia and astrocytes.
In addition, Sema4d was expressed on microglia, while PlexinB2 was expressed on astrocytes , Sema4D and PlexinB2 expressions were up-regulated in EAE model mice.
PlexinB2-positive astrocytes prefer to be close to Sema4d-positive microglia.
Further inhibiting the activities of Sema4D and PlexinB2 by CRISPR-Cas9 technology can effectively alleviate the symptoms of EAE model mice.
The researchers found out the treasure signal Sema4D-PlexinB2 through RABID-seq, and also found another pair of treasure signals EphB3-Ephrin-B3.
Erythropoietin human hepatocyte B (EphB) receptor is a member of the receptor tyrosine kinase family, and it works by binding to its ligand EphfinB.
The expression of EphB3-Ephrin-B3 on glial cells The expression of Efnb3 on microglia in EAE model mice increased, which promoted the inflammatory activation of microglia; the expression of EphB3 on astrocytes also increased.
After interfering RNA technology reduces the expression of Efnb3 on microglia and EphB3 on astrocytes, the inflammation activation state is significantly reduced.
Inhibition of EphB3 receptor can also significantly improve the symptoms of EAE model mice.
In summary, this article has discovered two signaling molecular pathways that "talk" between microglia and astrocytes through the development of a new cell-interacting single-cell sequencing technology RABID-seq: EphB3-Ephrin-B3 and Sema4D-PlexinB2.
[References] 1.
https://doi.
org/10.
1126/science.
abf1230 The pictures in the article are all from the references
