In Vitro Study of Notch1 Signaling in Cardiomyocyte Differentiation, Maturation, and Gene Expression Regulation

This study investigates the role of Notch1 signaling in cardiomyocyte differentiation, maturation, and gene expression regulation. Using neonatal mice with floxed Notch1 genes, Cre recombinase expression was controlled by MLC2v to achieve Notch1 knockout, supplemented by γ-secretase inhibitor DAPT and gene transfection to construct various cardiomyocyte models. Techniques such as immunofluorescence staining, Western blot, Myomesin staining, and quantitative RT-PCR were used to analyze Notch1 signaling activation and changes in differentiation-related genes Jagged1 and Hes1. Results revealed that inhibiting Notch1 signaling significantly promoted cardiomyocyte differentiation and maturation, upregulating cardiac differentiation markers Jagged1 and Hes1. Conversely, Notch1 overexpression enhanced signaling, inhibiting differentiation and downregulating related genes. Mechanical stretch enhanced Notch1 activation and inhibited differentiation, while NSAID intervention had opposite effects. These findings demonstrate the crucial role of Notch1 signaling in cardiomyocyte differentiation, with implications for cardiac development and disease research.