Manipulating the intracellular environment of hippocampal slices: pH and high-energy phosphates

Tim S. Whittingham, Eduardo Warman, Hussein Assaf, Thomas J. Sick, Joseph C. LaManna

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

The intracellular energetic environment of rat hippocampal slices was manipulated by bolstering ATP levels following the addition of adenosine to the incubation medium, or by manipulating intracellular pH. Addition of 8 mM adenosine to the incubation medium increased total tissue adenylate and ATP content, but did not prolong electrical function during anoxia. Further, it resulted in long-lasting alterations in normoxic evoked responses. Intracellular pH (pHi) was changed by manipulating the bicarbonate/CO2 ratio of the incubation medium, or by adding amiloride, a hydrogen/sodium antiport blocker. Estimates of intracellular pH using the creatine kinase equilibrium agree with those obtained by Neutral red scanning spectrophotometry in control conditions. However, only Neutral red indicated an acidification with amiloride treatment, while the creatine kinase equilibrium was preferentially affected by hypercapnia, suggesting the presence of at least two pH compartments in hippocampal brain slices. These manipulations cannot be carried out easily in vivo, and provide a means of determining the importance of metabolic changes on neural function during anoxia.

Original languageEnglish (US)
Pages (from-to)83-91
Number of pages9
JournalJournal of Neuroscience Methods
Volume28
Issue number1-2
DOIs
StatePublished - May 1989

Keywords

  • Adenosine
  • Adenosine triphosphate
  • Amiloride
  • Neutral red
  • pH
  • Phosphocreatine

ASJC Scopus subject areas

  • Neuroscience(all)

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