High-resolution 2-deoxyglucose autoradiography in quick-frozen slabs of neonatal rat olfactory bulb

Thane E. Benson, Gail D. Burd, Charles A. Greer, Dennis M.D. Landis, Gordon M. Shepherd

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

Abstract

We have used rapid freezing and freeze-substitution fixation to permit electron microscopic study of [3H]2-deoxyglycucose autoradiographs. The techniques minimize diffusion of label into processing fluids and, by inference, migration of label within tissue. Slabs of olfactory bulbs from 12-day-old rats were quick-frozen after one hour of exposure to physiological olfactory stimuli. In light microscopic autoradiographs at low magnification, the neuropil of individual olfactory glomeruli appeares uniformyly labeled with different levels by label clusters, suggesting greater deoxyglucose uptake by olfactory nerve terminals as compared with their postsynaptic dendrites. Periglomerular neurons were labeled differentially. Some microglia and glia precursor cells were heavily labeld in all bulbar laminae. The ultrastructure of cells and neuropil in all bulbar laminae was well-preserved. Cell processes and organelles could be identified in both stained sections and unstained electron microscopic autoradiographs. These experiments demonstrate the feasibility of combining quick-freezing with freeze substitution, in order to extend the resolution of studies using diffusable tracers such as 2-deoxyglucose. The results suggest that this is a promising method for assessing several controversies concerning deoxyglucose incorporation and neuronal and glial metabolism.

Original languageEnglish (US)
Pages (from-to)67-78
Number of pages12
JournalBrain Research
Volume339
Issue number1
DOIs
StatePublished - Jul 22 1985
Externally publishedYes

Keywords

  • 2-deoxyglucose
  • autoradiography
  • electron microscopy
  • glia
  • neurons
  • neuropil
  • olfactory bulb

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

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