Type 10 adenylyl cyclase mediates mitochondrial Bax translocation and apoptosis of adult rat cardiomyocytes under simulated ischaemia/reperfusion

Avinash Appukuttan, Ruhr-University Bochum, Germany
Sascha A. Kasseckert A. Sascha A. Kasseckert, University of Giessen, Giessen, Germany
Mustafa Micoogullari, University of Giessen, Giessen, Germany
Jan-Paul Flacke, Ruhr-University Bochum, Germany
Sanjeev Kumar, Ruhr-University Bochum, Germany
Alexandra Woste, University of Giessen, Giessen, Germany
Yaser Abdallah, Aga Khan University
Lutz Pott, Ruhr-University Bochum, Bochum, Germany
H. Peter Reusch, Ruhr-University Bochum, Bochum, Germany
Yury Ladilov, Ruhr-University Bochum, Bochum, Germany

This work was published before the author joined Aga Khan University.


Aims. Apoptosis of cardiomyocytes significantly contributes to the development of post-ischaemic cardiomyopathy. Although mitochondria have been suggested to play a crucial role in this process, the precise mechanisms controlling the mitochondria-dependent apoptosis in cardiomyocytes under ischaemia/reperfusion are still poorly understood. Here we aimed to analyse the role of the soluble adenylyl cyclase (sAC).

Methods and results Adult rat cardiomyocytes were subjected to simulated in vitro ischaemia (SI) consisting of glucose-free anoxia at pH 6.4. Apoptosis was detected by DNA laddering, chromatin condensation, and caspases cleavage. SI led to the translocation of sAC to the mitochondria and mitochondrial depolarization followed by cytochrome c release, caspase-9/-3 cleavage and apoptosis during simulated reperfusion (SR). Pharmacological inhibition of sAC during SI, but not during SR, significantly reduced the SI/SR-induced mitochondrial injury and apoptosis. Similarly, sAC knock-down mediated by an adenovirus coding for shRNA targeting sAC prevented the activation of the mitochondrial pathway of apoptosis. Analysis of the link between sAC and apoptosis revealed a sAC and protein kinase A-dependent Bax phosphorylation at Thr(167) and its translocation to mitochondria during SI, which subsequently caused mitochondrial oxygen radical formation followed by cytochrome c release and caspase-9 cleavage during SR.

Conclusion These results suggest a key role of sAC in SI-induced mitochondrial Bax translocation and activation of the mitochondrial pathway of apoptosis in adult cardiomyocytes.