A morphometric analysis of bipolar and multipolar TH-IR neurons treated with the neurotoxin MPP+ in co-cultures from mesencephalon and striatum of embryonic C57BL/6 mice.
Mesencephalic cultures contain two morphologically different tyrosine hydroxylase (TH)-immunoreactive (IR) neurons, fusiform bipolar, and pyramidal multipolar, which project to different anatomical structures (ventral striatum and neostriatum). The possibility of functional difference of these cells in Parkinson's disease led us study the effect of 1-methyl-4-phenylpyridinium (MPP+) on them. Survival and morphology of the two groups was studied in dissociated co-cultures from mesencephalon and striatum of embryonic C57BL/6 mice. Cells were grown at first in serum containing medium and then in serum free medium supplemented with hormones. Cultures were exposed to different concentrations of MPP+ on day 9 and 13 for 24 hr. They were fixed and stained with an anti-TH antibody. 0.1-1.0 microM MPP+ caused a dramatic reduction of the total area of TH-IR neurons. At 0.1 microM MPP+ some area was reduced, at 0.5 microM it appeared similar to controls, and decreased further at 1.0 microM. The relation of soma to total area showed that the decrease of the neuronal size was mainly due to the degeneration of the neuronal processes. The length of neuronal tree as well as the number of terminal segments were reduced dose dependently when cells were treated with the toxin. Similar results were obtained for bipolar and multipolar neurons. A significant difference in the decrease in total area was observed between the two age groups when cells were treated with MPP+, as older cells appeared to be more sensitive. When other parameters were checked no apparent difference was present.
Koutsilieri
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E
Institute of Medical Chemistry, Veterinary Medical University of Vienna, Austria.
Chen
T S
TS
Kruzik
P
P
Rausch
W D
WD
D015655:1-Methyl-4-phenylpyridinium / Q000494:pharmacology*; D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D003342:Corpus Striatum / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D008636:Mesencephalon / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D017966:Pyramidal Cells / Q000502:physiology; D014446:Tyrosine 3-Monooxygenase / Q000276:immunology*
D014446:Tyrosine 3-Monooxygenase; D015655:1-Methyl-4-phenylpyridinium