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Life science is the dorsal root ganglion (DRG) neuron, which is the somatosensory portal cell.
The specific ion channel it expresses controls the input and coding of many sensory information, including temperature, mechanical force, itch, etc.
, of which TRPV1 is the first The temperature-sensitive channel was identified in 1997, and PIEZO2 as a mechanically sensitive channel that mediates light touch and proprioception was identified in 2010.
Over the years, comprehensive studies on the structure and function of molecules such as TRPV1 and PIEZO have also been directly locked in 2021.
The Nobel Prize in Physiology or Medicine in 2015 reflects people’s attention to molecules that mediate mechanosensation
.
However, the mechanical sensitivity of DRG neurons exhibits diversity.
The analysis of current inactivation mechanics mainly shows three different speeds: rapidly adapting (RA), intermediately adapting (IA), and Slow or ultra-slow phase (SA/ultra SA)
.
At present, it is determined that PIEZO2 is the target channel of RA current, but the channel molecules that mediate the IA and SA currents are not clear
.
Recently, the team of Professor Bertrand Coste of the University of Aix-Marseille used patch-clamp sequencing (Patch-seq) to perform single-cell-scale cluster analysis of the transcription profiles of DRG neurons with different mechanically sensitive current dynamics.
The internal researchers provided a practical transcriptomics data set, which was published on November 2, 2021 in Cell Reports, a journal of Cell Press
.
Press and hold the picture to scan the QR code to read the paper.
Figure 1 Schematic diagram of the research strategy in this paper.
In order to study the molecular mechanism of the different mechanical sensitivity dynamics of DRG neurons, the author used whole-cell patch clamp electrophysiology to identify the mechanically sensitive current dynamics of DRG neurons.
, And then use electrodes to collect cells, so as to further obtain the single-cell transcriptomics data of the cell, so that the dynamics of the mechanically sensitive current and the gene transcription profile are matched, and these cells are classified according to their current dynamics, and the single-cell transcriptomics data is measured at the same time.
Transcriptomics of cells
.
The author successfully established a single-cell cDNA library for 62 DRG neurons typed by electrophysiological recording, and their current dynamics can fully cover these three mechanical sensitivity characteristics, and finally 53 suitable for single-cell RNA sequencing analysis were obtained.
For DRG neurons, it is worth mentioning that the patch-seq gene sequencing coverage rate of these cells is 13,359±1,478
.
Figure 2 Correspondence analysis of single-cell mechanically sensitive current characteristics and transcriptome characteristics of DRG neurons.
According to gene expression profiles, these DRG neurons that have identified mechanically sensitive current dynamics can be divided into 7 functional cell groups, which are highly consistent Mapped to a previous report from 1580 single-cell RNAseq data
.
These cells can be divided into NF (corresponding to the low-threshold mechano-receptor LTMR), NP1 (related to multimodal hyperalgesia), NP2 and NP3 (related to itching), and PEP1 (class C temperature hyperalgesia).
Related), PEP2 (mildly myelinated Aδ hyperalgesic receptor), TH (class C low-threshold mechanical receptor)
.
The expression profile analysis of this data set can see some classic DRG neuron marker genes and genes related to temperature and itch, which reflects the credible overall quality of the patch-seq data set of this work
.
Figure 3 Transcriptome cluster analysis of DRG neurons.
At the same time, some classically recognized and newly reported mechanosensitive channel coding genes are also clearly expressed in this data set
.
Figure 4 The expression of known or predicted mechanosensitive channel genes in this data set.
Finally, the author also analyzed the two recently reported mechanosensitive channel channel candidate genes TACAN (Tmem120a) and Tentonin-3 (Tmem150c) corresponding to the machine in the data set.
The expression of sensitive current characteristic DRG neuron type
.
The expression levels of the two are fairly uniform in all cells, which means that they are not related to the difference in mechanically sensitive current dynamics
.
Further RNAi knockdown experiments on DRG neurons showed that the silencing of these two genes did not significantly affect the magnitude of any type of mechanosensitive current
.
The author's data and some recent reports from other research groups show that Tmem120a and Tmem150c may not be the molecules responsible for IA and SA mechanically sensitive currents on DRG neurons
.
Figure 5 Tmem120a and Tmem150c do not participate in the DRG mechano-sensitive current.
To sum up, this work uses Patch-seq technology to make a single-cell-scale transcriptional spectrum annotation for the mechano-sensitive current dynamics of DRG neurons, providing researchers in the field An open data set for the study of candidate genes for the DRG neuron mechanical sensitive channel
.
Abstract: A variety of mechanosensory neurons are involved in somatic sensations such as touch, proprioception, and pain.
However, the molecular components of the mechanotransduction protein machinery that support these sensory modes remain to be discovered
.
In this paper, we combine the mechanosensory (MS) current recording of mechanosensory neurons with single-cell RNA sequencing: the transcription profile is mapped to previously determined sensory neuron types to identify transcription data sets and cell types Relevance
.
The correlation between the mechanically sensitive current characteristics and the single-cell transcriptome provides a one-to-one correspondence between the mechanical properties and the neuronal population defined by the transcriptome
.
In addition, the comparison of gene expression differences provides a set of candidate genes for the mechanical transduction complex
.
In line with expectations, Piezo2 genes were found to be enriched in fast-adapted MS current-expressing neurons; while Tmem120a and Tmem150c genes, which are believed to mediate slow-adapted MS currents, are evenly expressed in all mechanosensory neuron subtypes, and further The gene knockout experiment ruled out the possibility that these two genes are involved in mediating MS currents in sensory neurons
.
This data set also provides an open resource for further exploring the determinants of the type-specificity of mechanosensory cells
.
A variety of mechanosensory neurons are involved in touch, proprioception, and pain.
Many molecular components of the mechanotransduction machinery subserving these sensory modalities remain to be discovered.
Here, we combine recordings of mechanosensitive (MS) currents in mechanosensory neurons with single-cell RNA sequencing.
Transcriptional profiles are mapped onto previously identified sensory neuron types to identify cell-type correlates between datasets.
Correlation of current signatures with single-cell transcriptomes provides a one-to-one correspondence between mechanoelectric properties and transcriptomically defined neuronal populations.
Moreover, a gene-expression differential comparison provides a set of candidate genes for mechanotransduction complexes.
Piezo2 is expectedly found to be enriched in rapidly adapting MS current-expressing neurons, whereas Tmem120a and Tmem150c, thought to mediate slow- type MS currents, are uniformly expressed in all mechanosensory neuron subtypes.
Further knockdown experiments disqualify them as mediating MS currents in sensory neurons.
This dataset constitutes an open resource to explore further the cell-type-specific determinants of mechanosensory properties.
Scroll down to read the abstract original Chinese content is for reference only, please refer to the English original Researcher E-mail: wei_xiong@tsinghua.
edu.
cn researcher and doctoral supervisor of the School of Life Sciences, Tsinghua University, researcher of IDG/McGovern Institute for Brain Research, Tsinghua UniversityThis dataset constitutes an open resource to explore further the cell-type-specific determinants of mechanosensory properties.
Scroll down to read the abstract.
The original Chinese content is for reference only, and the English original text shall prevail -mail:wei_xiong@tsinghua.
edu.
cn, researcher and doctoral supervisor of the School of Life Sciences, Tsinghua University, researcher of IDG/McGovern Institute of Brain Research, Tsinghua UniversityThis dataset constitutes an open resource to explore further the cell-type-specific determinants of mechanosensory properties.
Scroll down to read the abstract.
The original Chinese content is for reference only, and the English original text shall prevail -mail:wei_xiong@tsinghua.
edu.
cn researcher and doctoral supervisor of the School of Life Sciences, Tsinghua University, researcher of IDG/McGovern Institute of Brain Research, Tsinghua University
.
Xiong Wei's laboratory mainly uses mouse genetics, molecular and cell biology, optical imaging, and electrophysiological methods to study the normal functioning of the auditory system and the biological mechanisms of abnormal auditory perception.
Related work was published in Nat Protoc.
, eLife as the corresponding author.
, PNAS and other academic journals
.
Dr.
Wei Xiong is a Principal Investigator in School of Life Sciences and IDG/McGovern Institute for Brain Research at Tsinghua University.
Xiong laboratory mainly use mouse genetics, molecular and cellular biology, optical imaging and electrophysiology to study mechanisms underlying normal hearing and deafness.
Dr.
Xiong has recently published researches from his laboratory in Journals including Nat.
Protoc.
, eLife, and PNAS.
Swipe down to read the English resume related paper information.
The specific ion channel it expresses controls the input and coding of many sensory information, including temperature, mechanical force, itch, etc.
, of which TRPV1 is the first The temperature-sensitive channel was identified in 1997, and PIEZO2 as a mechanically sensitive channel that mediates light touch and proprioception was identified in 2010.
Over the years, comprehensive studies on the structure and function of molecules such as TRPV1 and PIEZO have also been directly locked in 2021.
The Nobel Prize in Physiology or Medicine in 2015 reflects people’s attention to molecules that mediate mechanosensation
.
However, the mechanical sensitivity of DRG neurons exhibits diversity.
The analysis of current inactivation mechanics mainly shows three different speeds: rapidly adapting (RA), intermediately adapting (IA), and Slow or ultra-slow phase (SA/ultra SA)
.
At present, it is determined that PIEZO2 is the target channel of RA current, but the channel molecules that mediate the IA and SA currents are not clear
.
Recently, the team of Professor Bertrand Coste of the University of Aix-Marseille used patch-clamp sequencing (Patch-seq) to perform single-cell-scale cluster analysis of the transcription profiles of DRG neurons with different mechanically sensitive current dynamics.
The internal researchers provided a practical transcriptomics data set, which was published on November 2, 2021 in Cell Reports, a journal of Cell Press
.
Press and hold the picture to scan the QR code to read the paper.
Figure 1 Schematic diagram of the research strategy in this paper.
In order to study the molecular mechanism of the different mechanical sensitivity dynamics of DRG neurons, the author used whole-cell patch clamp electrophysiology to identify the mechanically sensitive current dynamics of DRG neurons.
, And then use electrodes to collect cells, so as to further obtain the single-cell transcriptomics data of the cell, so that the dynamics of the mechanically sensitive current and the gene transcription profile are matched, and these cells are classified according to their current dynamics, and the single-cell transcriptomics data is measured at the same time.
Transcriptomics of cells
.
The author successfully established a single-cell cDNA library for 62 DRG neurons typed by electrophysiological recording, and their current dynamics can fully cover these three mechanical sensitivity characteristics, and finally 53 suitable for single-cell RNA sequencing analysis were obtained.
For DRG neurons, it is worth mentioning that the patch-seq gene sequencing coverage rate of these cells is 13,359±1,478
.
Figure 2 Correspondence analysis of single-cell mechanically sensitive current characteristics and transcriptome characteristics of DRG neurons.
According to gene expression profiles, these DRG neurons that have identified mechanically sensitive current dynamics can be divided into 7 functional cell groups, which are highly consistent Mapped to a previous report from 1580 single-cell RNAseq data
.
These cells can be divided into NF (corresponding to the low-threshold mechano-receptor LTMR), NP1 (related to multimodal hyperalgesia), NP2 and NP3 (related to itching), and PEP1 (class C temperature hyperalgesia).
Related), PEP2 (mildly myelinated Aδ hyperalgesic receptor), TH (class C low-threshold mechanical receptor)
.
The expression profile analysis of this data set can see some classic DRG neuron marker genes and genes related to temperature and itch, which reflects the credible overall quality of the patch-seq data set of this work
.
Figure 3 Transcriptome cluster analysis of DRG neurons.
At the same time, some classically recognized and newly reported mechanosensitive channel coding genes are also clearly expressed in this data set
.
Figure 4 The expression of known or predicted mechanosensitive channel genes in this data set.
Finally, the author also analyzed the two recently reported mechanosensitive channel channel candidate genes TACAN (Tmem120a) and Tentonin-3 (Tmem150c) corresponding to the machine in the data set.
The expression of sensitive current characteristic DRG neuron type
.
The expression levels of the two are fairly uniform in all cells, which means that they are not related to the difference in mechanically sensitive current dynamics
.
Further RNAi knockdown experiments on DRG neurons showed that the silencing of these two genes did not significantly affect the magnitude of any type of mechanosensitive current
.
The author's data and some recent reports from other research groups show that Tmem120a and Tmem150c may not be the molecules responsible for IA and SA mechanically sensitive currents on DRG neurons
.
Figure 5 Tmem120a and Tmem150c do not participate in the DRG mechano-sensitive current.
To sum up, this work uses Patch-seq technology to make a single-cell-scale transcriptional spectrum annotation for the mechano-sensitive current dynamics of DRG neurons, providing researchers in the field An open data set for the study of candidate genes for the DRG neuron mechanical sensitive channel
.
Abstract: A variety of mechanosensory neurons are involved in somatic sensations such as touch, proprioception, and pain.
However, the molecular components of the mechanotransduction protein machinery that support these sensory modes remain to be discovered
.
In this paper, we combine the mechanosensory (MS) current recording of mechanosensory neurons with single-cell RNA sequencing: the transcription profile is mapped to previously determined sensory neuron types to identify transcription data sets and cell types Relevance
.
The correlation between the mechanically sensitive current characteristics and the single-cell transcriptome provides a one-to-one correspondence between the mechanical properties and the neuronal population defined by the transcriptome
.
In addition, the comparison of gene expression differences provides a set of candidate genes for the mechanical transduction complex
.
In line with expectations, Piezo2 genes were found to be enriched in fast-adapted MS current-expressing neurons; while Tmem120a and Tmem150c genes, which are believed to mediate slow-adapted MS currents, are evenly expressed in all mechanosensory neuron subtypes, and further The gene knockout experiment ruled out the possibility that these two genes are involved in mediating MS currents in sensory neurons
.
This data set also provides an open resource for further exploring the determinants of the type-specificity of mechanosensory cells
.
A variety of mechanosensory neurons are involved in touch, proprioception, and pain.
Many molecular components of the mechanotransduction machinery subserving these sensory modalities remain to be discovered.
Here, we combine recordings of mechanosensitive (MS) currents in mechanosensory neurons with single-cell RNA sequencing.
Transcriptional profiles are mapped onto previously identified sensory neuron types to identify cell-type correlates between datasets.
Correlation of current signatures with single-cell transcriptomes provides a one-to-one correspondence between mechanoelectric properties and transcriptomically defined neuronal populations.
Moreover, a gene-expression differential comparison provides a set of candidate genes for mechanotransduction complexes.
Piezo2 is expectedly found to be enriched in rapidly adapting MS current-expressing neurons, whereas Tmem120a and Tmem150c, thought to mediate slow- type MS currents, are uniformly expressed in all mechanosensory neuron subtypes.
Further knockdown experiments disqualify them as mediating MS currents in sensory neurons.
This dataset constitutes an open resource to explore further the cell-type-specific determinants of mechanosensory properties.
Scroll down to read the abstract original Chinese content is for reference only, please refer to the English original Researcher E-mail: wei_xiong@tsinghua.
edu.
cn researcher and doctoral supervisor of the School of Life Sciences, Tsinghua University, researcher of IDG/McGovern Institute for Brain Research, Tsinghua UniversityThis dataset constitutes an open resource to explore further the cell-type-specific determinants of mechanosensory properties.
Scroll down to read the abstract.
The original Chinese content is for reference only, and the English original text shall prevail -mail:wei_xiong@tsinghua.
edu.
cn, researcher and doctoral supervisor of the School of Life Sciences, Tsinghua University, researcher of IDG/McGovern Institute of Brain Research, Tsinghua UniversityThis dataset constitutes an open resource to explore further the cell-type-specific determinants of mechanosensory properties.
Scroll down to read the abstract.
The original Chinese content is for reference only, and the English original text shall prevail -mail:wei_xiong@tsinghua.
edu.
cn researcher and doctoral supervisor of the School of Life Sciences, Tsinghua University, researcher of IDG/McGovern Institute of Brain Research, Tsinghua University
.
Xiong Wei's laboratory mainly uses mouse genetics, molecular and cell biology, optical imaging, and electrophysiological methods to study the normal functioning of the auditory system and the biological mechanisms of abnormal auditory perception.
Related work was published in Nat Protoc.
, eLife as the corresponding author.
, PNAS and other academic journals
.
Dr.
Wei Xiong is a Principal Investigator in School of Life Sciences and IDG/McGovern Institute for Brain Research at Tsinghua University.
Xiong laboratory mainly use mouse genetics, molecular and cellular biology, optical imaging and electrophysiology to study mechanisms underlying normal hearing and deafness.
Dr.
Xiong has recently published researches from his laboratory in Journals including Nat.
Protoc.
, eLife, and PNAS.
Swipe down to read the English resume related paper information.
