Inositol 1,4,5-trisphosphate receptor 2 as a novel marker of vasculature to delineate processes of cardiopulmonary development
Inositol 1,4,5-trisphosphate receptor 2 as a novel marker of vasculature to delineate processes of cardiopulmonary development
复制标题
DOI：
10.1016/j.ydbio.2019.11.011
复制
发表时间：
2020-02-15
影响因子：
2.7
通讯作者：
Yamagishi, Hiroyuki
中科院分区：
文献类型：
--
作者：
Ishizaki-Asami, Reina;Uchida, Keiko;Yamagishi, Hiroyuki
先天性心脏病(CHDS)累及流出道(OFT)，如持续性动脉干(PTA)，会导致死亡率和发病率，不仅影响心脏，还影响肺血管系统。与CHD相关的PA疾病的发病机制和发病机制目前仍知之甚少，部分原因是PA发生的特定标记物还不存在。肌醇1，4，5-三磷酸受体的三种亚型(IP(3)R1、2和3)是许多组织和器官所必需的细胞内钙通道。我们发现，在转基因小鼠的发育过程中，IP(3)R2在血管和心脏中都有表达，其中一个LacZ标记基因被敲入了IP3R2基因座。整体和切片LacZ染色显示，IP(3)R2-LacZ阳性细胞仅见于PA的平滑肌细胞或中膜，并在发育过程中融合成αSMA阳性细胞。此外，我们的分析表明，从E13.5到E18.5，IP(3)R2-LacZ阳性的PA平滑肌层从PA中央逐渐延伸到外周PM，这支持了PA发生的远端血管生成学说，而IP(3)R2-LacZ在肺动脉干的平滑肌细胞中很少表达。将IP3R-LacZ小鼠与等位基因TBX1低构型的小鼠杂交，发现TBX1突变体的PTA可能是由于肺干发育不全或发育不全所致；因此，在这些突变体中，左右中央到外周的PM直接连接到动脉干的背侧。此外，我们还在TBX1突变的肺中发现了间质间充质细胞增多和肺内胚层成熟延迟。我们的研究确定IP(3)R2是一种新的标记物，可以清晰地显示PA发育过程中的PA，并可用于研究心肺发育和疾病。
Congenital heart diseases (CHDs) involving the outflow tract (OFT), such as persistent truncus arteriosus (PTA), lead to mortality and morbidity with implications not only in the heart, but also in the pulmonary vasculature. The mechanisms of pulmonary artery (PA) development and the etiologies underlying PA disorders associated with CHD remain poorly understood partly because of a specific marker for PA development is nonexistent. The three subtypes of inositol 1,4,5-trisphosphate receptors (IP(3)R1, 2, and 3) are intracellular Ca2+ channels that are essential for many tissues and organs. We discovered that IP(3)R2 was expressed in the vasculature and heart during development using transgenic mice, in which a LacZ marker gene was knocked into the IP3R2 locus. Whole-mount and section LacZ staining showed that IP(3)R2-LacZ-positive cells were detectable exclusively in the smooth muscle cells, or tunica media, of PA, merging into alpha SMA-positive cells during development. Furthermore, our analyses suggested that IP(3)R2-LacZ positive PA smooth muscle layers gradually elongate from the central PA to the peripheral PM from E13.5 to E18.5, supporting the distal angiogenesis theory for the development of PA, whereas IP(3)R2-LacZ was rarely expressed in smooth muscle cells in the pulmonary trunk. Crossing IP3R-LacZ mice with mice hypomorphic for Tbx1 alleles revealed that PTA of Tbx1 mutants may result from agenesis or hypoplasia of the pulmonary trunk; thus, the left and right central to peripheral PM connect directly to the dorsal side of the truncus arteriosus in these mutants. Additionally, we found hypercellular interstitial mesenchyme and delayed maturation of the lung endoderm in the Tbx1 mutant lungs. Our study identifies IP(3)R2 as a novel marker for clear visualization of PA during development and can be utilized for studying cardiopulmonary development and disease.