Rolfs Laura A, Falat Elizabeth J, Gutzman Jennifer H
Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI 53211, USA.
G3 (Bethesda). 2025 Jan 8;15(1). doi: 10.1093/g3journal/jkae260.
Non-muscle myosin (NMII) motor proteins have diverse developmental functions due to their roles in cell shape changes, cell migration, and cell adhesion. Zebrafish are an ideal vertebrate model system to study the NMII encoding myh genes and proteins due to high sequence homology, established gene editing tools, and rapid ex utero development. In humans, mutations in the NMII encoding MYH genes can lead to abnormal developmental processes and disease. This study utilized zebrafish myh9a, myh9b, and myh10 null mutants to examine potential genetic interactions and roles for each gene in development. It was determined that the myh9b gene is the most critical NMII encoding gene, as myh9b mutants develop pericardial edema and have a partially penetrant lethal phenotype, which was not observed in the other myh mutants. This study also established that genetic interactions occur between the zebrafish myh9a, myh9b, and myh10 genes where myh9b is required for the expression of both myh9a and myh10, and myh10 is required for the expression of myh9b. Additionally, protein analyses suggested that enhanced NMII protein stability in some mutant backgrounds may play a role in compensation. Finally, double mutant studies revealed different and more severe phenotypes at earlier time points than single mutants, suggesting roles for tissue specific genetic redundancy, and in some genotypes, haploinsufficiency. These mutants are the first in vivo models allowing for the study of complete loss of the NMIIA and NMIIB proteins, establishing them as valuable tools to elucidate the role of NMII encoding myh genes in development and disease.
非肌肉肌球蛋白(NMII)运动蛋白因其在细胞形状变化、细胞迁移和细胞黏附中的作用而具有多种发育功能。由于序列同源性高、已建立的基因编辑工具以及快速的子宫外发育,斑马鱼是研究编码NMII的肌球蛋白重链(myh)基因和蛋白质的理想脊椎动物模型系统。在人类中,编码MYH的NMII基因突变可导致异常的发育过程和疾病。本研究利用斑马鱼myh9a、myh9b和myh10基因敲除突变体来研究每个基因在发育中的潜在遗传相互作用和作用。研究确定myh9b基因是最关键的编码NMII的基因,因为myh9b突变体出现心包水肿并具有部分致死表型,而在其他myh突变体中未观察到这种情况。本研究还确定斑马鱼myh9a、myh9b和myh10基因之间存在遗传相互作用,其中myh9b是myh9a和myh10表达所必需的,而myh10是myh9b表达所必需的。此外,蛋白质分析表明,在某些突变背景下增强的NMII蛋白质稳定性可能在补偿中起作用。最后,双突变体研究揭示了在早期时间点比单突变体更不同且更严重的表型,表明组织特异性遗传冗余以及在某些基因型中的单倍剂量不足发挥了作用。这些突变体是首个用于研究NMIIA和NMIIB蛋白完全缺失的体内模型,使其成为阐明编码NMII的myh基因在发育和疾病中的作用的有价值工具。