Byerly Kathryn, Wolfe Cayla, Parris Hannah, Griggs Charlotte, Wilson Emily, Huff Matthew, Griggs Molly, Morningstar Jordan, Guo Lilong, Tang Fulei, Guz Jan, Petrucci Taylor, Phookan Ranan, Loizzi Brian, Gensemer Cortney, Norris Russell A
Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29407, USA.
Department of Comparative Medicine, Medical University of South Carolina, Charleston, SC 29407, USA.
Cells. 2025 May 24;14(11):774. doi: 10.3390/cells14110774.
Intercellular interactions among cardiac cell populations are essential for cardiac morphogenesis, yet the molecular mechanisms orchestrating these events remain incompletely understood. Dachsous1 (Dchs1), an atypical cadherin linked to mitral valve prolapse, is a core planar cell polarity protein whose function in the developing heart has not been fully elucidated. To address this, we generated a Dchs1-HA knock-in mouse model to define its spatial, temporal, and cellular expression patterns. Using immunohistochemistry, western blotting, and single-cell transcriptomics across developmental stages, we demonstrate that cardiac Dchs1 expression is restricted to non-cardiomyocyte lineages. DCHS1 displays dynamic subcellular localization and tissue organization depending on the developmental timepoint, with staining being found in epicardial and endocardial surfaces at earlier embryonic stages and in the compact myocardium in later fetal and neonatal stages. During fetal and neonatal stages, DCHS1-positive non-myocyte, non-endothelial cells form polarized extensions that bridge endothelial and non-myocyte, non-endothelial cells, suggesting direct heterotypic and homotypic interactions. Western blotting revealed evidence of DCHS1 proteolytic cleavage, with intracellular C-terminal fragments. RNA co-expression with its binding partner FAT4 supports a conserved, non-myocyte-specific DCHS1-FAT4 signaling axis. These findings identify DCHS1 as a potential molecular tether that is utilized in intercellular communications during cardiac development, with implications for congenital and acquired heart disease.
心脏细胞群体之间的细胞间相互作用对于心脏形态发生至关重要,然而协调这些事件的分子机制仍未完全了解。Dachsous1(Dchs1)是一种与二尖瓣脱垂相关的非典型钙黏蛋白,是一种核心平面细胞极性蛋白,其在发育中心脏中的功能尚未完全阐明。为了解决这个问题,我们生成了一个Dchs1-HA基因敲入小鼠模型,以确定其空间、时间和细胞表达模式。通过对不同发育阶段进行免疫组织化学、蛋白质印迹和单细胞转录组学分析,我们证明心脏Dchs1表达仅限于非心肌细胞谱系。DCHS1根据发育时间点表现出动态的亚细胞定位和组织排列,在胚胎早期阶段,在心外膜和心内膜表面有染色,而在胎儿后期和新生儿阶段,在致密心肌中有染色。在胎儿和新生儿阶段,DCHS1阳性的非心肌、非内皮细胞形成极化延伸,连接内皮细胞和非心肌、非内皮细胞,提示存在直接的异型和同型相互作用。蛋白质印迹显示有DCHS1蛋白水解切割的证据,存在细胞内C末端片段。与其结合伴侣FAT4的RNA共表达支持了一个保守的、非心肌细胞特异性的DCHS1-FAT4信号轴。这些发现确定DCHS1是心脏发育过程中细胞间通讯中潜在的分子连接物,对先天性和后天性心脏病具有重要意义。