Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN, 46202, USA.
Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
Sci Rep. 2023 Jul 1;13(1):10688. doi: 10.1038/s41598-023-36862-2.
The establishment of left-right patterning in mice occurs at a transient structure called the embryonic node or left-right organizer (LRO). Previous analysis of the LRO has proven challenging due to the small cell number and transient nature of this structure. Here, we seek to overcome these difficulties to define the transcriptome of the LRO. Specifically, we used single cell RNA sequencing of 0-1 somite embryos to identify LRO enriched genes which were compared to bulk RNA sequencing of LRO cells isolated by fluorescent activated cell sorting. Gene ontology analysis indicated an enrichment of genes associated with cilia and laterality terms. Furthermore, comparison to previously identified LRO genes identified 127 novel LRO genes, including Ttll3, Syne1 and Sparcl1, for which the expression patterns were validated using whole mount in situ hybridization. This list of novel LRO genes will be a useful resource for further studies on LRO morphogenesis, the establishment of laterality and the genetic causes of heterotaxy.
在小鼠中建立左右模式的过程发生在一个称为胚胎节点或左右组织者（LRO）的短暂结构中。由于该结构的细胞数量少且具有瞬态特性，因此之前对 LRO 的分析具有挑战性。在这里，我们试图克服这些困难来定义 LRO 的转录组。具体来说，我们使用 0-1 体节胚胎的单细胞 RNA 测序来鉴定 LRO 富集的基因，并将其与通过荧光激活细胞分选分离的 LRO 细胞的批量 RNA 测序进行比较。基因本体分析表明，与纤毛和左右性术语相关的基因富集。此外，与先前鉴定的 LRO 基因的比较确定了 127 个新的 LRO 基因，包括 Ttll3、Syne1 和 Sparcl1，其表达模式使用整体原位杂交进行了验证。该列表中的新 LRO 基因将成为进一步研究 LRO 形态发生、左右性建立以及异构性遗传原因的有用资源。
