Bousalis Deanna, Lacko Christopher S, Hlavac Nora, Alkassis Fariz, Wachs Rebecca A, Mobini Sahba, Schmidt Christine E, Kasahara Hideko
J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL, United States.
Department of Physiology and Functional Genomics, University of Florida, Gainesville, FL, United States.
Front Cardiovasc Med. 2020 May 29;7:93. doi: 10.3389/fcvm.2020.00093. eCollection 2020.
Congenital heart disease (CHD) affects almost one percent of all live births. Despite diagnostic and surgical reparative advances, the causes and mechanisms of CHD are still primarily unknown. The extracellular matrix plays a large role in cell communication, function, and differentiation, and therefore likely plays a role in disease development and pathophysiology. Cell adhesion and gap junction proteins, such as integrins and connexins, are also essential to cellular communication and behavior, and could interact directly (integrins) or indirectly (connexins) with the extracellular matrix. In this work, we explore disparities in the expression and spatial patterning of extracellular matrix, adhesion, and gap junction proteins between wild type and mutant mice. Decellularization and proteomic analysis, Western blotting, histology, immunostaining, and mechanical assessment of embryonic and neonatal wild type and mutant mouse hearts were performed. An increased abundance of collagen IV, fibronectin, and integrin β-1 was found in mutant neonatal mouse hearts, as well as increased expression of connexin 43 in embryonic mutant hearts. Furthermore, a ventricular noncompaction phenotype was observed in both embryonic and neonatal mutant hearts, as well as spatial disorganization of ECM proteins collagen IV and laminin in mutant hearts. Characterizing such properties in a mutant mouse model provides valuable information that can be applied to better understanding the mechanisms of congenital heart disease.
先天性心脏病（CHD）影响着近1%的活产婴儿。尽管在诊断和手术修复方面取得了进展，但CHD的病因和机制仍然主要不明。细胞外基质在细胞通讯、功能和分化中发挥着重要作用，因此可能在疾病发展和病理生理学中发挥作用。细胞粘附蛋白和间隙连接蛋白，如整合素和连接蛋白，对细胞通讯和行为也至关重要，并且可以直接（整合素）或间接（连接蛋白）与细胞外基质相互作用。在这项研究中，我们探讨了野生型和突变型小鼠之间细胞外基质、粘附和间隙连接蛋白在表达和空间模式上的差异。对胚胎期和新生期的野生型和突变型小鼠心脏进行了脱细胞和蛋白质组分析、蛋白质印迹、组织学、免疫染色以及力学评估。在突变型新生小鼠心脏中发现IV型胶原蛋白、纤连蛋白和整合素β-1的丰度增加，在胚胎期突变型心脏中连接蛋白43的表达也增加。此外，在胚胎期和新生期突变型心脏中均观察到心室肌小梁疏松的表型，并且在突变型心脏中细胞外基质蛋白IV型胶原蛋白和层粘连蛋白存在空间紊乱。在突变型小鼠模型中表征这些特性提供了有价值的信息，可用于更好地理解先天性心脏病的机制。
