Jun 14, 2023
Experimental design
- michela.deleidi 1,2,
- Bianca Marchetti3,4,
- Federico Bertoli1,2,
- Carmela Giachino3
- 1Mitochondria and Inflammation in Neurodegenerative Diseases, DZNE, Tübingen-Germany;
- 2Hertie Institute for Clinical Brain Research, University of Tübingen;
- 3Neuropharmacology Laboratory, Oasi Research Institute-IRCCS, Troina, Italy;
- 4Biomedical and Biotechnological Sciences, Pharmacology Section, University of Catania-Italy
Protocol Citation: michela.deleidi , Bianca Marchetti, Federico Bertoli, Carmela Giachino 2023. Experimental design. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwj7m2lmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 31, 2023
Last Modified: June 14, 2023
Protocol Integer ID: 79814
Keywords: Experimental design, experimental design the protocol detail, experimental design, protocol detail, protocol
Funders Acknowledgements:
ASAP-Aligning Science Across Parkinson’s
Grant ID: ASAP-000420
Abstract
The protocol details the experimental design.
Attachments
k4dqb48if.docx
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Materials
Materials
- lipopolysaccharide
- Escherichia coli serotype O111:B4) (Sigma-Aldrich)
- 0.9% sterile NaCl
Troubleshooting
Experimental design
WT and G2019S mice were exposed to intraperitoneal (ip) injections of a low dose of lipopolysaccharide LPS (0,1 mg kg-1 Escherichia coli serotype O111:B4) (Sigma-Aldrich), administrated twice a week for 12 weeks (Supplementary Figure 1). Treatments were performed in two different age groups, 3M (young adult) and 7M (at the start of middle-age). WT and G2019S mice exposed to ip injections of 0.9% sterile NaCl were used as controls.
Young mice receiving the treatment for 12 weeks were sacrificed at 6 M; middle aged mice were subdivided in 2 groups; one group received the treatment for 12 weeks was killed at 10 M, the second group received the treatment for 12 weeks and from 10 M to 16 M received saline only (Supplementary Figure 1A-B).
Mice (n = 20/experimental group) were randomly assigned to one of seven experimental conditions for each genotype:
1. 3 M Basal (no injections);
2. 6 M NaCl (NaCl injections started at 3 M);
3. 6 M LPS (LPS injections started at 3 M);
4.10 M NaCl (NaCl injections started at 7 M);
5.10 M LPS (injections started at 7M);
6. 16 M NaCl (injections started at 7 M);
7.16 M LPS (injections started at 7 M).
Clinical evaluation (body weight, mantel status, lethargy, reluctance to move, grooming behavior) was carried out weekly until sacrifice.
A second series of experiments were carried out (n=8 mice/experimental group) for immunohistochemical and protein analyses.