Ultrastructural localization of flotillin-1 to cholesterol-rich membrane microdomains, rafts, in rat brain tissue

There is much interest in research on cholesterol-rich membrane microdomains, rafts, in the field of neurobiology. However, no one has shown the ultrastructure of rafts in tissues. We examined the ultrastructure of rafts in rat brain tissue by pre-embedding immunoelectron microscopy using flotillin-...

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Bibliographic Details
Published in:Brain research 2003-03, Vol.965 (1), p.83-90
Main Authors: Kokubo, Hideko, Helms, J.Bernd, Ohno-Iwashita, Yoshiko, Shimada, Yukiko, Horikoshi, Yuko, Yamaguchi, Haruyasu
Format: Article
Language:English
Subjects:
Animals
BCθ
Brain - ultrastructure
Brain Chemistry
Brain tissue
Cell Membrane - chemistry
Cell Membrane - ultrastructure
Cholesterol - analysis
Cholesterol-rich membrane microdomain
Female
Flotillin-1
Intracellular Membranes - chemistry
Intracellular Membranes - ultrastructure
Membrane Microdomains - chemistry
Membrane Microdomains - ultrastructure
Membrane Proteins - analysis
Membrane Proteins - ultrastructure
Organelles - chemistry
Organelles - ultrastructure
Raft
Rats
Rats, Wistar
Ultrastructure
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Summary:There is much interest in research on cholesterol-rich membrane microdomains, rafts, in the field of neurobiology. However, no one has shown the ultrastructure of rafts in tissues. We examined the ultrastructure of rafts in rat brain tissue by pre-embedding immunoelectron microscopy using flotillin-1 antibody, which is a biochemical marker of lipid rafts, and BCθ, which is nicked and biotinylated θ-toxin, and binds to membrane cholesterol of rafts. Flotillin-1- and BCθ-labeled areas were patchy and prominent on the plasma membranes of small processes and synapses in the neuropil. The size of flotillin-1 labeling was 40–200 nm. In addition, the membrane of lysosome and Golgi apparatus were frequently labeled for flotillin-1 with a patchy pattern. Flotillin-1 and BCθ were mostly colocalized in double immunolabeling on a part of the plasma membranes of small processes and secondary lysosome membranes. We first indicate that flotillin-1 localizes to BCθ-positive cholesterol-rich membrane microdomains in vivo, and that flotillin-1 and BCθ could be ultrastructural raft markers in neural tissue.
ISSN:0006-8993
1872-6240
DOI:10.1016/S0006-8993(02)04140-9