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 light-sensitive (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.
Bibliographical noteFunding Information:
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.
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. 2-AG, 2-arachidonoylglycerol; ACSF, artificial cerebral spinal fluid; AMPA, ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid; CCA, canonical correlation analysis; DA, dopamine; DEG, differentially expressed gene; FPKM, fragments per kilobase million; GO, gene ontology; IEG, immediate-early gene; LGN, lateral geniculate nucleus; LS, light-sensitive neuron; mACh, muscarinic acetylcholine; mAChR, muscarinic acetylcholine receptors; nACh, nicotinic acetylcholine; NMDA, N-methyl-D-aspartate; NS, non-light-sensitive neuron; PCA, principal component analyses; PMT, photomultiplier tube; PV, parvalbumin; RGC, retinal ganglion cell; scRNA-seq, single cell RNA sequencing; SST, somatostatin; t-SNE, t-distributed Stochastic Neighbor Embedding; V1, primary visual cortex; vivo -seq, in vivo single cell RNA-seq Jianwei Liu, Mengdi Wang, Le Sun, Na Clara Pan, Changjiang Zhang, Junjing Zhang, Zhentao Zuo, Sheng He, Qian Wu, and Xiaoqun Wang declare that they have no conflict of interest. All institutional and national guidelines for the care and use of laboratory animals were followed.
© 2020, The Author(s).
- calcium imaging in vivo
- light sensitivity
- whole cell recording in vivo
PubMed: MeSH publication types
- Journal Article
- Research Support, Non-U.S. Gov't