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| Item Type: | Article |
|---|---|
| Title: | FRET based ratiometric Ca(2+) imaging to investigate immune-mediated neuronal and axonal damage processes in experimental autoimmune encephalomyelitis |
| Creators Name: | Siffrin, V., Birkenstock, J., Luchtmann, D.W., Gollan, R., Baumgart, J., Niesner, R.A., Griesbeck, O. and Zipp, F. |
| Abstract: | BACKGROUND: Irreversible axonal and neuronal damage are the correlate of disability in patients suffering from multiple sclerosis (MS). A sustained increase of cytoplasmic free [Ca2+] is a common upstream event of many neuronal and axonal damage processes and could represent an early and potentially reversible step. NEW METHOD: We propose a method to specifically analyze the neurodegenerative aspects of experimental autoimmune encephalomyelitis by Forster Resonance Energy Transfer (FRET) imaging of neuronal and axonal Ca2+ dynamics by two-photon laser scanning microscopy (TPLSM). RESULTS: Using the genetically encoded Ca2+ sensor TN-XXL expressed in neurons and their corresponding axons, we confirm the increase of cytoplasmic free [Ca2+] in axons and neurons of autoimmune inflammatory lesions compared to those in non-inflamed brains. We show that these relative [Ca2+] increases were associated with immune-neuronal interactions. COMPARISON WITH EXISTING METHODS: In contrast to Ca2+-sensitive dyes the use of a genetically encoded Ca2+ sensor allows reliable intraaxonal free [Ca2+] measurements in living anesthetized mice in health and disease. This method detects early axonal damage processes in contrast to e.g. cell/axon morphology analysis, that rather detects late signs of neurodegeneration. CONCLUSIONS: Thus, we describe a method to analyze and monitor early neuronal damage processes in the brain in vivo. |
| Keywords: | EAE/MS, Two-Photon Laser Scanning Microscopy, Intravital Microscopy, FRET, Ca2+ Imaging Neurodegeneration, Animals, Mice |
| Source: | Journal of Neuroscience Methods |
| ISSN: | 0165-0270 |
| Publisher: | Elsevier |
| Volume: | 249 |
| Page Range: | 8-15 |
| Date: | 15 July 2015 |
| Official Publication: | https://doi.org/10.1016/j.jneumeth.2015.04.005 |
| PubMed: | View item in PubMed |
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