Changes in subtypes of Ca microdomains following partial injury to the central nervous system

I. Lozić, Carole Bartlett, Jeremy Shaw, Killugudi Iyer, Sarah Dunlop, Matt Kilburn, Melinda Fitzgerald

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)
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Rapid changes in Ca2+ concentration and location in response to injury play key roles in a range of biological systems. However, quantitative analysis of changes in size and distribution of Ca2+ microdomains in specific cell types in whole tissue samples has been limited by analytical resolution and reliance on indirect Ca2+ indicator systems. Here, we combine the unique advantages of nanoscale secondary ion mass spectrometry (NanoSIMS) with immunohistochemistry to directly quantify changes in number, size and intensity of Ca microdomains specific to axonal or glial regions vulnerable to spreading damage following neurotrauma. Furthermore, using NanoSIMS allows separate quantification of Ca microdomains according to their co-localization with areas enriched in P. We rapidly excise and cryopreserve optic nerve segments from adult rat at time points ranging from 5 minutes to 3 months after injury, allowing assessment of Ca microdomains dynamics with minimal disruption due to tissue processing. We demonstrate significantly more non-P co-localized Ca microdomains in glial than axonal regions in normal optic nerve. The density of Ca microdomains not co-localized with areas enriched in P rapidly, selectively and significantly decreases after injury; densities of Ca microdomains co-localized with P enriched areas are unchanged. An efflux of Ca2+ from microdomains not co-localized with P may contribute to the structural and functional deficits observed in nerve vulnerable to spreading damage following neurotrauma. NanoSIMS analyses of Ca microdomains allow quantitative and novel insights into Ca dynamics, applicable to a range of normal, as well as diseased or injured mammalian systems. © 2014 The Royal Society of Chemistry.
Original languageEnglish
Pages (from-to)455-464
Issue number3
Early online date17 Dec 2013
Publication statusPublished - Mar 2014


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