Mengdi Wang
Le Sun
Na Clara Pan
Changjiang Zhang
Junjing Zhang
Zhentao Zuo
Sheng He
Qian Wu
Xiaoqun Wang
1 State Key Laboratory of Brain and Cognitive Science, CAS Center for Excellence in Brain Science and Intelligence Technology, Institute of Brain-Intelligence Technology(Shanghai), Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;
2 State Key Laboratory of Cognitive Neuroscience and Learning, Beijing Normal University, Beijing 100875, China;
3 University of Chinese Academy of Sciences, Beijing 100049, China;
4 Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China;
5 IDG/McGovern Institute for Brain Research, Beijing Normal University, Beijing 100875, China;
6 Advanced Innovation Center for Human Brain Protection, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China
Funds: This work was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB32010100), National Basic Research Program of China (2019YFA0110101, 2017YFA0103303, 2017YFA0102601), the National Natural Science Foundation of China (NSFC) (31671072, 31771140, 81891001), the Beijing Brain Initiative of Beijing Municipal Science & Technology Commission (Z181100001518004), Open Research Fund of the State Key Laboratory of Cognitive Neuroscience and Learning.
Received Date: 2019-10-07
Rev Recd Date:2020-01-09
Abstract
Abstract
Vision formation is classically based on projections from retinal ganglion cells (RGC) to the lateral geniculate nucleus (LGN) and the primary visual cortex (V1). Neurons in the mouse V1 are tuned to light stimuli. Although the cellular information of the retina and the LGN has been widely studied, the transcriptome profiles of single light-stimulated neuron in V1 remain unknown. In our study, in vivo calcium imaging and whole-cell electrophysiological patch-clamp recording were utilized to identify 53 individual cells from layer 2/3 of V1 as lightsensitive (LS) or non-light-sensitive (NS) by single-cell light-evoked calcium evaluation and action potential spiking. The contents of each cell after functional tests were aspirated in vivo through a patch-clamp pipette for mRNA sequencing. Moreover, the three-dimensional (3-D) morphological characterizations of the neurons were reconstructed in a live mouse after the whole-cell recordings. Our sequencing results indicated that V1 neurons with a high expression of genes related to transmission regulation, such as Rtn4r and Rgs7, and genes involved in membrane transport, such as Na+/K+ ATPase and NMDA-type glutamatergic receptors, preferentially responded to light stimulation. Furthermore, an antagonist that blocks Rtn4r signals could inactivate the neuronal responses to light stimulation in live mice. In conclusion, our findings of the vivo-seq analysis indicate the key role of the strength of synaptic transmission possesses neurons in V1 of light sensory.Keywords: light sensitivity,
vivo-seq,
patch-seq,
calcium imaging in vivo,
whole cell recording in vivo
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