Dendrite loss is a characteristic early indicator of toxin-induced neurodegeneration in rat midbrain slices

Exp Neurol. 2000 Jan;161(1):306-16. doi: 10.1006/exnr.1999.7259.

Abstract

In rat brain substantia nigra catecholamine neurons in vitro, a sensitive indicator of excitatory amino-acid-induced damage is dendritic degeneration that precedes the loss of the cell body. The present study has shown that dendritic loss is not specific for excitatory amino acids and is an early indicator of neurodegeneration produced by numerous agents that initiate damage by different primary cellular actions. Rats were anesthetised by fluothane inhalation and killed, and the brain was rapidly removed. Three-hundred-micrometer-thick slices containing substantia nigra were incubated for 2 h at 35 degrees C in the presence or absence of kainic acid (50 microM), 1-methyl-4-phenylpyridinium ion (10 or 50 microM), ouabain (10 or 30 microM), 6-hydroxydopamine (10 or 100 microM), potassium cyanide (100 microM or 1 mM), or elevated extracellular potassium chloride (25, 50, or 100 mM). The slices were fixed and recut into thin sections (30 micrometer) and substantia nigra dopamine neurons were immunolabeled for tyrosine hydroxylase coupled to diaminobenzidine. Both the cell body and the extensive dendritic projections were immunolabeled. Each agent caused a similar pattern of toxicity including loss of tyrosine-hydroxylase-immunolabeled dendrites at lower concentrations and damage to, or disintegration of, the cell bodies at higher concentrations. For example, 100 microM potassium cyanide reduced the proportion of substantia nigra neurons which exhibited dendrites from 66 +/- 4% (SEM) in controls to 54 +/- 7%, without obvious changes in cell bodies. After 1 mM potassium cyanide, only 13 +/- 2% of substantia nigra neurons retained dendrites and cell bodies were shrunken or disintegrated. Loss of dendrites was also evident in substantia nigra neurons stained with cresyl violet or immunolabeled for microtubule-associated protein 2. The findings suggest that disruption of the dendritic arbor is an early indicator of neurodegeneration, irrespective of how this is initiated. The approach that we have developed may therefore prove valuable in investigating the mechanisms of degeneration of catecholamine neurons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzoxazines
  • Biomarkers
  • Cell Death
  • Coloring Agents
  • Dendrites / chemistry
  • Dendrites / enzymology
  • Dendrites / pathology*
  • Immunohistochemistry
  • Male
  • Microtubule-Associated Proteins / analysis
  • Nerve Degeneration / chemically induced
  • Nerve Degeneration / pathology*
  • Neurotoxins
  • Organ Culture Techniques
  • Oxazines
  • Oxidopamine
  • Poisons
  • Potassium Cyanide
  • Rats
  • Rats, Sprague-Dawley
  • Substantia Nigra / pathology*
  • Sympatholytics
  • Tyrosine 3-Monooxygenase / analysis

Substances

  • Benzoxazines
  • Biomarkers
  • Coloring Agents
  • Microtubule-Associated Proteins
  • Neurotoxins
  • Oxazines
  • Poisons
  • Sympatholytics
  • microtubule-associated protein 1B
  • cresyl violet
  • Oxidopamine
  • Tyrosine 3-Monooxygenase
  • Potassium Cyanide