Neurons containing retrogradely transported Fluoro-Gold exhibit a variety of lysosomal profiles: a combined brightfield, fluorescence, and electron microscopic study.
The advantages of axonally transported Fluoro-Gold as a retrograde fluorescent marker are numerous. The objective of the present study was to determine whether transported Fluoro-Gold is visible in either semi-thin sections for light microscopy or thin sections for electron microscopy. Rats received injections of Fluoro-Gold into either the striatum or thoracic spinal cord. After appropriate survival times, labelled neurons were observed with the fluorescence microscope in brain regions that are known to project to the injected areas. Sections that contained labelled cells were embedded in plastic and examined with a fluorescence microscope. Semi-thin sections of unosmicated tissue displayed high-resolution fluorescent labelling of somata and dendrites. In contrast, osmicated tissue did not fluoresce, but numerous dark granules were observed in the dendritic and perikaryal cytoplasm of labelled neurons in toluidine blue stained sections that were examined with brightfield optics. The unosmicated tissue did not display these granules, and this finding suggested that the granules are composed of membranes. Neurons in other brain regions that are known not to project to the injection sites did not contain these dark granules. Adjacent thin sections examined with the electron microscope displayed numerous electron-dense, lysosome-like organelles in the cytoplasm of labelled neurons. The electron density of these organelles was greater than that of lysosomes in unlabelled neurons. Three types of distinctive organelles were observed in these preparations: (1) relatively dense concentric lamellar bodies of various sizes; (2) heterogeneous or lipofuscin-like lysosomes; and (3) coarse grained lysosomes. Control sections and unlabelled neurons did not display these organelles. Therefore, these organelles appear to correlate with Fluoro-Gold localized within the somata and dendrites of retrogradely labelled neurons. It is not known if they are the Fluoro-Gold itself, or represent a physiological effect on membranes. The results of this study indicate that Fluoro-Gold may be useful for tract tracing at the electron microscopic level.
Schmued
L C
LC
Department of Anatomy and Neurobiology, University of California, Irvine 92717.
Kyriakidis
K
K
Fallon
J H
JH
Ribak
C E
CE
D001692:Biological Transport; D005456:Fluorescent Dyes / Q000008:administration & dosage; D007031:Hypothalamus / Q000166:cytology / Q000648:ultrastructure; D007278:Injections, Spinal; D008247:Lysosomes / Q000648:ultrastructure*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology* / Q000648:ultrastructure; D012012:Red Nucleus / Q000166:cytology / Q000648:ultrastructure; D012154:Reticular Formation / Q000166:cytology; D013003:Somatosensory Cortex / Q000166:cytology / Q000648:ultrastructure; D013266:Stilbamidines; D013378:Substantia Nigra / Q000166:cytology / Q000648:ultrastructure
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D005456:Fluorescent Dyes; D013266:Stilbamidines