基于单细胞转录组技术解析泌乳奶牛瘤胃上皮组织细胞类型及功能

瘤胃上皮组织结构与功能在维持奶牛健康和泌乳性能中发挥着重要作用，而其组织内部的细胞异质性及分子特征有待深入探究。本研究以泌乳奶牛为对象，首先借助不同组织解离设备和条件，探索并建立符合大规模单细胞捕获追踪的瘤胃上皮组织充分解离和单细胞悬液制备方法；以此为基础，利用单细胞转录组技术研究泌乳奶牛瘤胃上皮组织细胞类型和基因表达特征，分析不同细胞类型间的互作机制，通过机器学习算法推拟细胞发育轨迹；采用RNA荧光原位杂交和免疫荧光手段对关键亚型的决定因子进行验证，同时揭示关键亚型在上皮组织中的空间定位和分布，基本阐明泌乳奶牛瘤胃上皮细胞类型及亚型的分布和功能。研究结果可为精确解析泌乳奶牛瘤胃上皮组织生理功能提供理论依据，对靶向制定营养调控策略促进瘤胃发育和改善瘤胃健康具有重要指导意义。

The complete structure and function of the rumen epithelium play vital roles in maintenance of the health and lactation performance of dairy cows. It is warranted to investigate the cellular heterogeneity and its molecular characteristics within the rumen epithelial tissues. In this proposed study, we intend to explore and establish a method for dissociation of rumen epithelial tissue and preparation of single-cell suspension, which are suitable for capturing and tracking of a large number of single cells. Single-cell transcriptome technology will then be conducted to investigate the cell types and their gene expression patterns in rumen epithelial tissues from lactating dairy cows. We will analyze the interactions between cell types and predict the cell developmental trajectory using machine learning algorithms. In addition, RNA fluorescence in situ hybridization and immunofluorescence are used to validate the driving factors of key cell subtypes and uncover the spatial localization of key subtypes in rumen epithelial tissues. The final goal of this proposal is to reveal the cell types, subtype distribution and function of rumen epithelial tissue in lactating dairy cows. The research results will provide theoretical evidence for accurate interpretation of physiological functions of rumen epithelial tissues of lactating dairy cows, and thus have important guiding significance in developing targeted nutritional strategies to improve rumen development and health.