Extracellular somatostatin measured by microdialysis in the hippocampus of freely moving rats: evidence for neuronal release

J Neurochem. 1993 Feb;60(2):671-7. doi: 10.1111/j.1471-4159.1993.tb03200.x.

Abstract

Intracerebral microdialysis combined with a sensitive and specific radioimmunoassay was used to monitor the neuronal release of somatostatin (somatostatin-like immunoreactivity, SLI) in the dorsal hippocampus of freely moving rats. The sensitivity of the radioimmunoassay was optimized to detect < 1 fmol/ml. The basal concentration of SLI in 20-min dialysate fractions (5 microliters/min) collected 24 h after probe implantation was stable over at least 200 min. The spontaneous efflux dropped by 54 +/- 6.4% (p < 0.05) when Ca2+ was omitted and 1 mM EGTA added to the Krebs-Ringer solution and by 65.5 +/- 3.2% (p < 0.05) in the presence of 1 microM tetrodotoxin. Depolarizing concentrations of the Na+ channel opener veratridine (6.25, 25, 100 microM) induced 11 +/- 2 (p < 0.05), 17 +/- 2 (p < 0.05), and 21 +/- 5 (p < 0.01) fold increase in SLI concentration, respectively, during the first 20 min of perfusion. The effect of 100 microM veratridine was blocked by coperfusion with 5 microM tetrodotoxin (p < 0.01) and reduced by 79% (p < 0.01) in the virtual absence of Ca2+. Neuronal depolarization by 20 min of perfusion with Krebs-Ringer solution containing 25 and 50 mM KCl and proportionally lowered Na+ increased the dialysate SLI 4.4 +/- 1 (p < 0.05) and 17 +/- 3 (p < 0.01) fold baseline, respectively. Ten micromolar ouabain, a blocker of Na+,K(+)-ATPase, increased the dialysate SLI 15-fold baseline, on average (p < 0.05), during 80 min of perfusion. The results demonstrate the suitability of brain microdialysis for monitoring the neuronal release of SLI and for studying its role in synaptic transmission.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / pharmacology
  • Dialysis / methods
  • Egtazic Acid / pharmacology
  • Hippocampus / drug effects
  • Hippocampus / metabolism*
  • Male
  • Microchemistry
  • Neurons / drug effects
  • Neurons / metabolism*
  • Ouabain / pharmacology
  • Potassium Chloride / pharmacology
  • Radioimmunoassay
  • Rats
  • Rats, Sprague-Dawley
  • Somatostatin / metabolism*
  • Tetrodotoxin / pharmacology
  • Veratridine / pharmacology

Substances

  • Tetrodotoxin
  • Somatostatin
  • Egtazic Acid
  • Ouabain
  • Potassium Chloride
  • Veratridine
  • Calcium