Simultaneous assessment of ecto- and cytosolic-5′-nucleotidase activities in brain micropunches

Miroslaw Mackiewicz, Jonathan D. Geiger, Allan I. Pack

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

We propose a new methodology for simultaneous assessment of ecto- and cytosolic-5′-nucleotidase that can be utilized in brain to measure the activity of these enzymes in micropunches of tissues. It is based on the differential sensitivity of both enzymes to α, β-methyleneadenosine 5′-diphosphate (AMP-CP) and the requirements for magnesium as a co-factor. The design of assay protocol contains an internal validation by allowing comparisons between total level of 5′-nucleotidase activity with that calculated from the sum of individual activities of the ecto- and cytosolic-5′-nucleotidases. We have applied this new approach to assess the activity of ecto- and cytosolic-5′-nucleotidase in the brain regions relevant to sleep regulation. The level of both enzymes was significantly lower in the cerebral cortex than other brain regions tested.

Original languageEnglish (US)
Pages (from-to)9-18
Number of pages10
JournalJournal of Neuroscience Methods
Volume104
Issue number1
DOIs
StatePublished - Dec 15 2000

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Keywords

  • 5′-Nucleotidase
  • Adenosine
  • Enzymatic assay
  • Sleep regulation

ASJC Scopus subject areas

  • Neuroscience(all)

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