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Electron microscopic cytochemical correlates of histofluorescence
Brain Research Bulletin
|July 1, 1982
Summary
This study introduces a new electron microscopic technique for precisely localizing biogenic amines like norepinephrine and dopamine in squirrel monkey tissues. The method reveals previously undescribed large deposits, enhancing our understanding of amine distribution in nervous systems.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Biogenic amines, including norepinephrine and dopamine, are crucial neurotransmitters.
- Accurate localization of these amines is essential for understanding neural function.
- Previous methods had limitations in fine structural localization and morphology preservation.
Purpose of the Study:
- To develop and validate a novel electron microscopic cytochemical technique for localizing biogenic amines.
- To identify and characterize the ultrastructural localization of norepinephrine and dopamine in specific squirrel monkey tissues.
- To compare electron microscopic findings with established histofluorescence methods.
Main Methods:
- Utilized a specific cytochemical technique involving a chromium (Cr) complex with an isoquinoline derivative.
- Formed amine-glutaraldehyde-Cr reaction product from unsubstituted biogenic amines and glutaraldehyde.
- Identified the reaction product using energy dispersive X-ray analysis and correlated Cr-positive sites with glyoxylic acid histofluorescence.
- Employed F-12 buffer perfusion prior to fixation for optimal tissue preservation.
Main Results:
- Successfully localized norepinephrine and dopamine electron microscopically in the substantia nigra, pineal body, adrenal medulla, and stellate ganglion of squirrel monkeys.
- Observed significantly large structures containing Cr-positive amine material, not previously described.
- Demonstrated a strong correlation between the size and location of these deposits and histofluorescence findings.
- Achieved excellent tissue morphology and fine structural localization.
Conclusions:
- The developed electron microscopic cytochemical technique provides precise localization of biogenic amines.
- The identification of novel large amine-containing structures advances the understanding of amine storage and distribution.
- This technique is valuable for studying biogenic amines in nervous tissue where fine structural detail and morphology are critical.