Role of the nucleotides cyclic ADP-ribose, ADP-ribose and NAADP in local and global Ca2+ signaling in mouse cardiac myocytes
Principal investigator:
Andreas H. Guse,
Institute of Biochemistry and Molecular Biology I: Cellular Signal Transduction, UKE
Co-workers partly involved:
Ralf Fliegert
Andreas Gasser
Susan Schlegel
Contact:
guse@uke.uni-hamburg.de
Objectives
To analyze the physiological role of novel calcium signalling systems in cardiac myocytes:
- the cyclic ADP-ribose/calcium signaling pathway,
- the NAADP/calcium signaling pathway,
- ADPR/TRPM2/calcium signaling pathway .
Questions
- Do extracellular signals change intracellular concentrations of cADPR, NAADP or ADPR in cardiac myocytes?
- Which role do the messengers cADPR and NAADP play in shaping the spatio-temporal calcium signaling pattern in cardiac myocytes?
- Does ADPR play a role for calcium signaling in cardiac myocytes?
Materials
- Freshly prepared cardiac myocytes
- Analogues of second messengers
Methods
- Confocal and conventional calcium imaging
- Calcium imaging combined with microinjection or patch-clamp recordings in isolated cells
- Mass analysis of second messengers by HPLC or enzymatic assays
- Expression analysis (RT-PCR, SDS-PAGE and western blot)
