Journal
MICROSCOPY RESEARCH AND TECHNIQUE
Volume 67, Issue 3-4, Pages 175-195Publisher
WILEY-LISS
DOI: 10.1002/jemt.20197
Keywords
cylindrical filaments with periodic thin regions (CF-PT); cylindrical filaments(CF); 2.0 tau polymer-like filaments
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Funding
- NIA NIH HHS [AG11054, AG05892] Funding Source: Medline
- NINDS NIH HHS [NS 18105] Funding Source: Medline
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Isolated paired helical filaments (PHFs) were visualized on glutaraldehyde vaportreated thin similar to 10-nm thick indirect carbon films using high-resolution transmission electron microscopy (TEM) and the negative stain, phosphotungstate acid (PTA) at near neutral pH of 6.8. PHF preparations were prepared with and without 1 minute of sonication. These same PHF were also deglycosylated with endoglycosidase F/N-glycosidase F for 1 hour or the PHF were dephosphorylated with PP-2A for 1 hour. The negatively stained PHF filaments were quantitatively studied by measuring their wide regions (W) their thin regions (T) and their helical turn period (L) and these separate parameters were averaged for each filament. In the unsonicated PHF preparation there were PHF, cylindrical filaments with periodic thin regions (CF-PT), cylindrical filaments (CF), as well as 2.0-nm tau polymer-like filaments. The CF-PT were characterized by W, T, and L measurements and the CF were characterized by diameter measurements. The paired helical filament model proposed by Kidd (1963, Nature 197:192-193) of two similar to 10 nm filaments twisting around each other every similar to 80 nm with a thin region of 10 nm and a wide region of 25 nm does not correspond to the PHF structures found. None of the PHF we observed were composed of a pair of filaments and all of the PHF appear to be a single filament. The wide regions ranged from 12.5-27 nm and the thin regions ranged from 4.5-12.3 nm. The helical turn periods ranged from 76-85 nm and were generally about 80 rim. Only the helical turn period of similar to 80 nm was a common property of the whole family of PHF structures. The CF-PT appear to be a PHF precursor filament. Deglycosylation of the PHF and CF-PT reduced their sizes by 0.5-0.6 nm and 0.7-1.0 nm, respectively, and the right-hand helicity of the PHF was lost after deglycosylation. Dephosphorylation with PP-2A reduced the PHF wide regions by 6.0 nm and the thin regions by 2.6 nm.
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