Autonomous activity and autophosphorylation of CAMPK-II in rat hippocampal slices: Effects of tissue preparation

I. Lengyel, M. Cammarota, V. A. Brent, J. A P Rostas*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)149-154
Number of pages6
JournalJournal of Neurochemistry
Volume76
Issue number1
DOIs
Publication statusPublished - 2001
Externally publishedYes

Keywords

  • Calcium
  • Calmodulin kinase
  • Freeze-thaw
  • Hippocampal slices
  • Methodology
  • Protein phosphorylation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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