Introduction
Methods
Cell culture and cell lines
Transgenic animals
Human tissue
Isolation of mouse brain
Immunohistochemistry of mouse tissue
Isolation of brain regions from mouse and human brain slices
Paraffin embedding, sectioning, and tissue preparation
Tissue homogenization
Transduction into biosensor cell lines
Inoculation of strains into transgenic mice
Generation of strains in secondary cells
Immunocytochemistry of secondary cell lines
Flow cytometry and analysis of seeding activity
Statistical analysis
Results
Fixed tissue retains tau seeding activity
Quantification of seeding activity in fixed tissue from PS19 mice
Age at Collection |
N (Total) | Male | Female |
---|---|---|---|
P301S Mice | |||
1 month | 6 | 3 | 3 |
2 month | 6 | 3 | 3 |
3 month | 6 | 3 | 3 |
4 month | 6 | 3 | 3 |
5 month | 6 | 3 | 3 |
6 month | 6 | 3 | 3 |
7 month | 6 | 3 | 3 |
8 month | 6 | 3 | 3 |
9 month | 6 | 3 | 3 |
10 month | 6 | 3 | 3 |
11 month | 6 | 3 | 3 |
12 month | 6 | 3 | 3 |
WT Mice | |||
1 month | 4 | 3 | 1 |
2 month | 1 | 0 | 1 |
5 month | 1 | 0 | 1 |
6 month | 1 | 1 | 0 |
9 month | 1 | 1 | 0 |
10 month | 1 | 0 | 1 |
12 month | 1 | 0 | 1 |
Quantification of spreading tau pathology
Time Course |
N (Total) | Male | Female | Average Age at Surgery (Days) | Average Age at Collection (Days) |
---|---|---|---|---|---|
3 weeks | |||||
DS1 | 2 | 0 | 2 | 94 | 115 |
DS9 | 4 | 0 | 4 | 90 | 111 |
DS10 | 4 | 0 | 4 | 91 | 112 |
6 weeks | |||||
DS1 | 3 | 0 | 3 | 77 | 119 |
DS9 | 4 | 1 | 3 | 80 | 122 |
DS10 | 4 | 0 | 4 | 80 | 122 |
12 weeks | |||||
DS1 | 2 | 0 | 2 | 71 | 155 |
DS9 | 4 | 2 | 2 | 70 | 154 |
DS10 | 3 | 1 | 2 | 71 | 155 |
Fixed tissue retains strain-specific conformations
Quantification of seeding activity in fixed tissue from the human brain
Patient Case | NFT/Aβ/α-synuclein Stage | M/F | Age | Year of formalin fixation | Notes |
---|---|---|---|---|---|
1 | a/0/0 | M | 31 | 1989 | Control |
2 | 1b/0/0 | M | 42 | 1989 | Control |
3 | III/0/0 | F | 90 | 1992 | No clinical AD diagnosis |
4 | III/2/0 | M | 65 | 1989 | No clinical AD diagnosis |
5 | V/3/0 | M | 88 | 1990 | Clinical AD |