目的检测弹性蛋白在大鼠心肌细胞和H9C2心肌细胞系内的表达。方法取新生和成年SD大鼠各5例的新鲜心脏,冷冻切片;取20只新生3 d SD大鼠心脏,通过0.05%胰蛋白酶和0.075%Ⅱ型胶原酶消化法,获取原代心肌细胞;免疫组织化学和免疫荧光技术检...目的检测弹性蛋白在大鼠心肌细胞和H9C2心肌细胞系内的表达。方法取新生和成年SD大鼠各5例的新鲜心脏,冷冻切片;取20只新生3 d SD大鼠心脏,通过0.05%胰蛋白酶和0.075%Ⅱ型胶原酶消化法,获取原代心肌细胞;免疫组织化学和免疫荧光技术检测大鼠心肌组织、原代心肌细胞和H9C2心肌细胞系的弹性蛋白表达;ELISA法检测培养的H9C2心肌细胞系上清中的弹性蛋白。结果在大鼠心肌细胞、H9C2心肌细胞系和新生、成年大鼠心肌组织上均发现心肌肌钙蛋白T和弹性蛋白的共表达。免疫组织化学结果发现,弹性蛋白在成纤维细胞、平滑肌细胞和细胞间质内均有表达。H9C2心肌细胞培养液的上清中检测到少量的弹性蛋白。结论大鼠心肌细胞表达弹性蛋白,可能参与心肌细胞的势能和细胞间质弹性纤维的构建。展开更多
The T-tubule (TT) system forms the structural basis for excitation-contraction coupling in heart and muscle cells. The morphogenesis of the TT system is a key step in the maturation of heart cells because it does not ...The T-tubule (TT) system forms the structural basis for excitation-contraction coupling in heart and muscle cells. The morphogenesis of the TT system is a key step in the maturation of heart cells because it does not exist in neonatal cardiomyocytes. In the present study, we quantified the morphological changes in TTs during heart cell maturation and investigated the role of junctophilin-2 (JP2), a protein known to anchor the sarcoplasmic reticulum (SR) to TT, in changes to TT morphological parameters. Analysis of confocal images showed that the transverse elements of TTs increased, while longitudinal elements decreased during the maturation of TTs. Fourier transform analysis showed that the power of ~2 m spatial components increased with cardiomyocytes maturation. These changes were preceded by increased expression of JP2, and were reversed by JP2 knockdown. These findings indicate that JP2 is required for the morphogenesis of TTs during heart development.展开更多
文摘目的检测弹性蛋白在大鼠心肌细胞和H9C2心肌细胞系内的表达。方法取新生和成年SD大鼠各5例的新鲜心脏,冷冻切片;取20只新生3 d SD大鼠心脏,通过0.05%胰蛋白酶和0.075%Ⅱ型胶原酶消化法,获取原代心肌细胞;免疫组织化学和免疫荧光技术检测大鼠心肌组织、原代心肌细胞和H9C2心肌细胞系的弹性蛋白表达;ELISA法检测培养的H9C2心肌细胞系上清中的弹性蛋白。结果在大鼠心肌细胞、H9C2心肌细胞系和新生、成年大鼠心肌组织上均发现心肌肌钙蛋白T和弹性蛋白的共表达。免疫组织化学结果发现,弹性蛋白在成纤维细胞、平滑肌细胞和细胞间质内均有表达。H9C2心肌细胞培养液的上清中检测到少量的弹性蛋白。结论大鼠心肌细胞表达弹性蛋白,可能参与心肌细胞的势能和细胞间质弹性纤维的构建。
基金supported by the National Basic Research Program of China (2011CB809101)the National Natural Science Foundation of China (30730013)
文摘The T-tubule (TT) system forms the structural basis for excitation-contraction coupling in heart and muscle cells. The morphogenesis of the TT system is a key step in the maturation of heart cells because it does not exist in neonatal cardiomyocytes. In the present study, we quantified the morphological changes in TTs during heart cell maturation and investigated the role of junctophilin-2 (JP2), a protein known to anchor the sarcoplasmic reticulum (SR) to TT, in changes to TT morphological parameters. Analysis of confocal images showed that the transverse elements of TTs increased, while longitudinal elements decreased during the maturation of TTs. Fourier transform analysis showed that the power of ~2 m spatial components increased with cardiomyocytes maturation. These changes were preceded by increased expression of JP2, and were reversed by JP2 knockdown. These findings indicate that JP2 is required for the morphogenesis of TTs during heart development.
