The coordinated spatial and temporal regulation of gene expression in the murine hindlimb determines the identity of mesenchymal progenitors and the development of diversity of musculoskeletal tissues they form. Hindlimb development has historically been studied with lineage tracing of individual genes selected a priori, or at the bulk tissue level, which does not allow for the determination of single cell transcriptional programs yielding mature cell types and tissues. To identify the cellular trajectories of lineage specification during limb bud development, we used single cell mRNA sequencing (scRNA-seq) to profile the developing murine hindlimb between embryonic days (E)11.5-E18.5. We found cell type heterogeneity at all time points, and the expected cell types that form the mouse hindlimb. In addition, we used RNA fluorescence in situ hybridization (FISH) to examine the spatial locations of cell types and cell trajectories to understand the ancestral continuum of cell maturation. This data provides a resource for the transcriptional program of hindlimb development that will support future studies of musculoskeletal development and generate hypotheses for tissue regeneration.

小鼠后肢基因表达的协调空间和时间调节决定了间充质祖细胞的身份及其形成的肌肉骨骼组织多样性的发展。历史上，后肢发育是通过先验选择的单个基因的谱系追踪或在大量组织水平上进行研究的，这不允许确定产生成熟细胞类型和组织的单细胞转录程序。为了确定肢芽发育过程中谱系规范的细胞轨迹，我们使用单细胞 mRNA 测序 (scRNA-seq) 来分析胚胎日 (E)11.5-E18.5 之间发育中的小鼠后肢。我们发现所有时间点的细胞类型异质性，以及形成小鼠后肢的预期细胞类型。此外，我们使用 RNA 荧光原位杂交 (FISH) 来检查细胞类型和细胞轨迹的空间位置，以了解细胞成熟的祖先连续体。这些数据为后肢发育的转录程序提供了资源，将支持未来的肌肉骨骼发育研究并产生组织再生的假设。