Pprotein and Cell . . . Wang Xiaoqun/Wu Wei collaborated on the Vivo-seq technique and revealed the molecular properties of the V1 epithelial photosensitive neurons in mice.

Vision is one of the direct means for animals to obtain external information.in mammals, the visual system contains many visual areas, and they occupy most of the cerebral cortex.at present, most of the researches are focused on the visual circuit, in which different types of neurons respond to light stimulation and their characteristics.however, the molecular characteristics of neurons in the visual loop that respond to various light stimuli are not clear.single cell RNA sequencing technology developed in recent years makes it possible to analyze the molecular characteristics of these neurons.in recent years, Wang Xiaoqun, Institute of Biophysics, Chinese Academy of Sciences, has revealed the dynamic changes of cell types and transcriptional regulation mechanism in the development of prefrontal cortex and hippocampus (Zhong et al., nature, 2018; (nature | Wang Xiaoqun / Wu Qian cooperates to decode the cellular and molecular mechanisms of hippocampal development and formation, a key brain area for human learning and memory).the researchers developed the technology of vivo SEQ by combining the technology with neuron function detection.this technology is of great significance for the study of transcriptome information of neurons with specific functions.recently, protein & amp; cell published a research paper entitled "integrated analysis of in vivo recording with single cell RNA SEQ data revisits molecular properties of light sensitive neurons in mouse V1".this work has established a new technology and method of vivo SEQ, which integrates the calcium imaging technology caused by stimulation, whole cell electrophysiological recording of living mice, single cell mRNA sequencing and three-dimensional morphological reconstruction of neurons based on two-photon microscopy.and applied this technique to analyze the responsive (LS) and non responsive (NS) neurons in the 2 / 3 layer of mouse cortex, comprehensively revealed the physiological, morphological and molecular characteristics of mouse V1 cortical light sensitive neurons, and revealed the genes directly related to the light response function in the neurons.in this study, the calcium signal changes of each cell and the action potential recorded by patch clamp were recorded. A total of 53 ls and NS neurons in the 2 / 3 layer of V1 cortex were obtained. At the same time, single cell transcriptome sequencing and morphological analysis were performed on these neurons.through the three-dimensional reconstruction of some neurons, it was found that there was no significant difference in the control range and dendrite complexity between ls and NS cells. However, there were significant differences in the peak value of EPSC and IPSC between the two types of neurons.single cell sequencing also indicated that compared with NS neurons, LS neurons were overexpressed of ion and synaptic transmission related genes, such as RTN4R and rgs7, and transmembrane transport related genes such as Na + / K + ATPase and NMDA type glutamatergic receptor.moreover, blocking RTN4R signaling pathway with blockers can make V1 neurons which are originally responsive to light become insensitive to light, and the response can be recovered after the antagonist elution, indicating that the gene plays an important role in the specific function of neurons.this study demonstrated that this vivo SEQ method can find the function, morphology and transient transcriptome characteristics of single cells in living animals.therefore, this research method provides a breakthrough in the functional characteristics and transcriptome characteristics of connective neurons.figure: vivo SEQ model diagram. It is reported that the research work was completed by the Institute of Biophysics, Chinese Academy of Sciences and Beijing Normal University.Dr. Liu Jianwei, associate researcher sun Le, engineer Wang Mengdi and Dr. pan Na of Institute of Biophysics, Chinese Academy of Sciences, are co authors of this paper. (1) Zhong s, Ding W, sun L, Lu y, Dong h, fan x, Liu Z, Chen R, Zhang s, Ma Q, Tang F, Wu Q, Wang Q, Tang F, Wu Q, Wang x, Wang X. December the development of the human hippocampus. Nature. 2020 Jan; 577 (7791): 531-5362. Zhong s, Zhang s, fan x, Wu Q, Yan L, Dong J, Zhang h, Li L, sun L, sun L, sun L, sun L, sun L, sun L, sun L, sun L, sun L, sun L, sun L, sun L, Dong J, Zhang h, Li L, sun L, sun L, sun L, Dong J, Zhang h, Li L, sun L, sun L, sun L, Dong J, Zhang h, Li L, sun L, sun L, sun L, sun L, Dong J L, pan n, Xu x, Tang F, Zhang J, Qiao J, Wang X. A single-cell RNA-seq survey of the developmental landscape of the human prefrontal cortex. Nature. 2018 Mar 22;555(7697):524-528.