Ball Kristen, Kinne Renee, Aguirre Aitor
Department of Biomedical Engineering, Michigan State University; Institute for Quantitative Health Sciences and Engineering, Michigan State University.
Institute for Quantitative Health Sciences and Engineering, Michigan State University.
J Vis Exp. 2020 Mar 10(157). doi: 10.3791/60926.
Congenital heart defects (CHD) are the most common type of birth defect in humans, affecting up to 1% of all live births. However, the underlying causes for CHD are still poorly understood. The developing mouse constitutes a valuable model for the study of CHD, because cardiac developmental programs between mice and humans are highly conserved. The protocol describes in detail how to produce mouse embryos of the desired gestational stage, methods to isolate and preserve the heart for downstream processing, quantitative methods to identify common types of CHD by histology (e.g., ventricular septal defects, atrial septal defects, patent ductus arteriosus), and quantitative histomorphometry methods to measure common muscular compaction phenotypes. These methods articulate all the steps involved in sample preparation, collection, and analysis, allowing scientists to correctly and reproducibly measure CHD.
先天性心脏缺陷(CHD)是人类最常见的出生缺陷类型,影响高达1%的活产婴儿。然而,CHD的潜在原因仍知之甚少。发育中的小鼠是研究CHD的宝贵模型,因为小鼠和人类之间的心脏发育程序高度保守。该方案详细描述了如何产生所需妊娠阶段的小鼠胚胎、分离和保存心脏以供下游处理的方法、通过组织学鉴定常见CHD类型(如室间隔缺损、房间隔缺损、动脉导管未闭)的定量方法,以及测量常见心肌致密化表型的定量组织形态计量学方法。这些方法阐明了样本制备、收集和分析所涉及的所有步骤,使科学家能够正确且可重复地测量CHD。