Impairment of mitochondrial calcium handling in a mtSOD1 cell culture model of motoneuron disease
1 Center of Physiology, Georg-August University, Goettingen, Germany
2 Department of Neurology, Georg-August University, Goettingen, Germany
3 Laboratory of Neurochemistry, Fondazione S. Lucia IRCCS, Rome, Italy
4 Department of Biology, University of Rome "Tor Vergata", Rome, Italy
5 Department of Physics, Georg-August University, Goettingen, Germany
6 Department of Geriatrics, Evangelisches Krakenhaus, Goettingen-Weende, Germany
BMC Neuroscience 2009, 10:64 doi:10.1186/1471-2202-10-64Published: 22 June 2009
Additional file 1:
Schematic representation of method and spectra. Schematic representation of the (A) CCD-imaging setup used for calcium imaging and (B) simultaneous measurements of [Ca2+]i and [Ca2+]mito in SH-SY5Y cell line preparations. C) Spectral view of calcium imaging. The fluorescence excitation at 510 nm and emission detected at 340 nm are shown for Ca2+-saturated (A) and Ca2+-free (B) Fura-2 in pH 7.2 buffer. D) Spectral view of simultaneous [Ca2+]i and [Ca2+]mito measurements generated by the Molecular Probes spectral view program. Fluorescence excitation of fura-2 was done at 390 nm and 550 nm of rhod-2, respectively; emission was at 510 nm (fura-2, solid line) and at 600 nm (rhod-2) separated by a 565 nm dichroic mirror.
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Additional file 2:
Mitochondria-dependent responses of cytosolic calcium in SH-SY5Y neuroblastoma cells (non-transfected parental cell line) loaded with Fura-2 AM and superfused with DMEM medium. A) Representative CCD camera photomicrograph of Fura-2 AM loaded SH-SY5Y cells. Scale bar is 10 μm. B) FCCP-evoked mitochondria-dependent responses of cytosolic calcium.
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