- Title
- Autonomous activity and autophosphorylation of CAMPK-II in rat hippocampal slices: effects of tissue preparation
- Creator
- Lengyel, I.; Cammarota, M.; Brent, V. A.; Rostas, J. A. P.
- Relation
- Journal of Neurochemistry Vol. 76, Issue 1, p. 149-154
- Publisher Link
- http://dx.doi.org/10.1046/j.1471-4159.2001.00058.x
- Publisher
- Blackwell Publishing Ltd
- Resource Type
- journal article
- Date
- 2001
- Description
- Measurement of the proportion of calcium/calmodulin-stimulated protein kinase II (CaMPK-II) that is autonomously active or phosphorylated on Thr286 is thought to provide an index of the degree to which CaMPK-II in a tissue has been activated. We have examined how various ways of handling hippocampal tissue can alter these properties. Both autonomous activity and phospho-Thr²⁸⁶ content was high in freshly dissected hippocampus or freshly cut hippocampal slices. After incubation of hippocampal slices in artificial cerebrospinal fluid for 120 min, both properties of CaMPK-II decreased to a steady state level. Freeze–thaw or cutting the equilibrated slices could rapidly increase both autonomous activity and phospho-Thr²⁸⁶ immunoreactivity of CaMPK-II. These increases were comparable to changes induced by experimental treatment. Therefore, our results suggest that considerable care needs to be taken over the way in which hippocampal slices are handled.
- Subject
- calcium; calmodulin kinase; freeze-thaw; hippocampal slices; methodology; protein phosphorylation
- Identifier
- http://hdl.handle.net/1959.13/26737
- Identifier
- uon:1080
- Identifier
- ISSN:0022-3042
- Language
- eng
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