多通道Patch-seq探究视皮层PVALB神经元在弱视中的生理和分子机制

弱视是一种严重影响视觉功能的儿童眼病，且发病机制复杂。视皮层的PVALB抑制性神经元介导视觉信息传递和眼优势柱可塑性，但其在弱视中的作用及机制尚未明确。我们前期利用单眼剥夺法构建了Pvalb-Cre/Ai9小鼠弱视模型，并运用多通道膜片钳联合单细胞转录组测序（Patch-seq）技术收集了PVALB神经元及周围神经元的转录组、电生理特征、突触连接、细胞形态和解剖位置等多模态数据。数据整合分析发现弱视相关的基因和信号通路与神经元的形态结构、电生理功能和可塑性显著相关。据此我们提出科学假说：PVALB抑制性神经元的分子、形态、生理和功能异常介导弱视的发生和发展。本项目拟继续利用多通道Patch-seq技术解析PVALB神经元在弱视发病过程中的多模态数据特征，并在分子遗传-形态位置-电生理特性-环路功能四位一体诠释PVALB神经元在弱视中的作用机理，为弱视患者的临床诊疗提供可靠的实验和理论依据。

Amblyopia is a pediatric eye disease that seriously affects visual function, and its pathogenesis is complex. PVALB inhibitory neurons in the visual cortex mediate visual information transmission and dominant column plasticity, but its role and mechanism in amblyopia remain unclear. In previous work, we constructed a Pvalb-Cre/Ai9 mouse amblyopia model through monocular deprivation, and collected multimodal data of PVALB neurons and related neurons, including transcriptomes, electrophysiological characteristics, synaptic connections, cell morphology and anatomical position with multi-patch-seq. Data integration analysis found that key genes and signaling pathways related to amblyopia are closely related to neuron electrophysiological properties, cell morphology and plasticity. Therefore, we hypothesized that molecular, morphological, physiological and functional abnormalities of PVALB inhibitory neurons mediate the occurrence and development of amblyopia. This project intends to use the multi-patch-seq technology to explore the multimodal data characteristics of PVALB neurons at the single-cell level during the onset of amblyopic disease. Thus, the functional mechanism of PVALB neurons in amblyopia is explained in terms of molecular genetics, morphological position, electrophysiological characteristics and loop function, providing reliable experimental and theoretical basis for clinical diagnosis and treatment of amblyopia patients.