Markandeya Yogananda S, Phelan Laura J, Woon Marites T, Keefe Alexis M, Reynolds Courtney R, August Benjamin K, Hacker Timothy A, Roth David M, Patel Hemal H, Balijepalli Ravi C
From the Cellular and Molecular Arrhythmia Research Program, Department of Medicine, University of Wisconsin, Madison, Wisconsin 53706.
the Veterans Affairs San Diego Healthcare Systems, San Diego, California 92161, and the Department of Anesthesiology, University of California at San Diego, La Jolla, California 92161.
J Biol Chem. 2015 Sep 4;290(36):22085-100. doi: 10.1074/jbc.M115.674945. Epub 2015 Jul 13.
Pathological cardiac hypertrophy is characterized by subcellular remodeling of the ventricular myocyte with a reduction in the scaffolding protein caveolin-3 (Cav-3), altered Ca(2+) cycling, increased protein kinase C expression, and hyperactivation of calcineurin/nuclear factor of activated T cell (NFAT) signaling. However, the precise role of Cav-3 in the regulation of local Ca(2+) signaling in pathological cardiac hypertrophy is unclear. We used cardiac-specific Cav-3-overexpressing mice and in vivo and in vitro cardiac hypertrophy models to determine the essential requirement for Cav-3 expression in protection against pharmacologically and pressure overload-induced cardiac hypertrophy. Transverse aortic constriction and angiotensin-II (Ang-II) infusion in wild type (WT) mice resulted in cardiac hypertrophy characterized by significant reduction in fractional shortening, ejection fraction, and a reduced expression of Cav-3. In addition, association of PKCα and angiotensin-II receptor, type 1, with Cav-3 was disrupted in the hypertrophic ventricular myocytes. Whole cell patch clamp analysis demonstrated increased expression of T-type Ca(2+) current (ICa, T) in hypertrophic ventricular myocytes. In contrast, the Cav-3-overexpressing mice demonstrated protection from transverse aortic constriction or Ang-II-induced pathological hypertrophy with inhibition of ICa, T and intact Cav-3-associated macromolecular signaling complexes. siRNA-mediated knockdown of Cav-3 in the neonatal cardiomyocytes resulted in enhanced Ang-II stimulation of ICa, T mediated by PKCα, which caused nuclear translocation of NFAT. Overexpression of Cav-3 in neonatal myocytes prevented a PKCα-mediated increase in ICa, T and nuclear translocation of NFAT. In conclusion, we show that stable Cav-3 expression is essential for protecting the signaling mechanisms in pharmacologically and pressure overload-induced cardiac hypertrophy.
病理性心脏肥大的特征是心室肌细胞的亚细胞重塑,伴有支架蛋白小窝蛋白-3(Cav-3)减少、钙(Ca2+)循环改变、蛋白激酶C表达增加以及钙调神经磷酸酶/活化T细胞核因子(NFAT)信号通路的过度激活。然而,Cav-3在病理性心脏肥大中调节局部Ca2+信号的确切作用尚不清楚。我们使用心脏特异性过表达Cav-3的小鼠以及体内和体外心脏肥大模型,来确定Cav-3表达在预防药理学和压力超负荷诱导的心脏肥大中的必要条件。野生型(WT)小鼠的横向主动脉缩窄和血管紧张素-II(Ang-II)输注导致心脏肥大,其特征为缩短分数、射血分数显著降低以及Cav-3表达减少。此外,在肥大的心室肌细胞中,PKCα与1型血管紧张素-II受体和Cav-3的结合被破坏。全细胞膜片钳分析表明,肥大的心室肌细胞中T型钙电流(ICa,T)表达增加。相比之下,过表达Cav-3的小鼠表现出对横向主动脉缩窄或Ang-II诱导的病理性肥大的保护作用,伴有ICa,T的抑制以及完整的Cav-3相关大分子信号复合物。在新生心肌细胞中,siRNA介导的Cav-3敲低导致PKCα介导的ICa,T的Ang-II刺激增强,进而导致NFAT的核转位。在新生心肌细胞中过表达Cav-3可防止PKCα介导的ICa,T增加和NFAT的核转位。总之,我们表明稳定的Cav-3表达对于保护药理学和压力超负荷诱导的心脏肥大中的信号机制至关重要。