Marker for evaluating or assisting in evaluating aging of hippocampus tissues
A hippocampal, tissue technology, applied in the field of biomedicine, which can solve problems such as heavy burden on individuals and society
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Embodiment 1
[0085] Example 1. Detection of β-galactosidase activity in aging hippocampus
[0086] 1. Hippocampus collection
[0087] Cynomolgus monkeys were anesthetized and perfused with saline, and after perfusion was complete and brain tissue blanched, the right hemisphere of each brain was fixed in 4% paraformaldehyde for histological analysis. Separate the left hemisphere of each brain according to brain region and freeze in liquid nitrogen. The hippocampus is located in the medial temporal lobe. Hippocampal tissues were carefully picked out on ice, sequentially divided into several sections, placed in cryovials, and stored in liquid nitrogen (limited technique when isolating cryopreserved tissue for the beginning of the experiment, one Only female young individuals were excluded).
[0088] The hippocampus tissue of 8 young cynomolgus monkeys and the hippocampus tissue of 8 old cynomolgus monkeys were selected as the experimental objects.
[0089] 2. Preparation of frozen section...
Embodiment 2
[0110] Example 2, Detection of Aging Hippocampal Aggresome
[0111] 1. Preparation of paraffin sections of hippocampal tissue
[0112] 1. The hippocampal tissue fixed in 4% paraformaldehyde solution (g:ml, mass volume ratio, solute is disodium hydrogen phosphate and sodium dihydrogen phosphate, solvent is ultrapure water) in a series of gradient alcohols (70%- 100%) in dehydration;
[0113] 2. Embed the tissue in paraffin;
[0114] 3. Use a rotary microtome to cut into a thickness of 5 μm;
[0115] 4. Place the slices on a microscope slide, dry at 56°C for 2 hours, and store at room temperature (RT).
[0116] The aforementioned hippocampus tissues were obtained from 8 young cynomolgus monkey individuals and 8 old cynomolgus monkey individuals respectively, and the specific collection method was the same as that in Example 1.
[0117] 2. Aggresome staining of hippocampal tissue
[0118] 1, with 100%, 100% and 100% (volume percentage composition) xylene and 100%, 100%, 90%,...
Embodiment 3
[0128] Example 3. Detection of molecular markers γH2A.X, HP1γ, β-Amyloid (4G8), dsDNA, H3K9me3 or LINE-1ORF2-p in aging hippocampal tissue
[0129] 1. Immunohistochemical staining of hippocampal tissue
[0130] 1. use 100%, 100% and 100% (volume percentage composition) xylene and 100%, 100%, 90%, 90%, 80%, 70% (volume percentage composition) gradient ethanol successively to hippocampus Tissue paraffin sections were dewaxed and hydrated;
[0131] 2. Place the slices in 1x citrate buffer, pH 6.0 (purchased from Zhongshan Jinqiao, Cat. No. ZLI-9065) and heat 5 times, 3 minutes each time, for antigen retrieval;
[0132] 3. After cooling to room temperature, rinse the slices 3 times in 1xPBS solution (pH value 7.3, PBS powder purchased from Zhongshan Jinqiao, Cat. No. ZLI-9063);
[0133] 4. Permeabilize with 0.4% (volume percentage, diluted in 1xPBS solution (pH value is 7.3, PBS powder purchased from Zhongshan Jinqiao, product number ZLI-9063)) Triton X-100 solution for 2 hours;...
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