Sep 29, 2025
Mouse studies
- Maria Jose Perez J.1
- 1Institut Imagine
- Team Deleidi
Protocol Citation: Maria Jose Perez J. 2025. Mouse studies . protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlkyn21g5r/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: September 29, 2025
Last Modified: September 29, 2025
Protocol Integer ID: 228449
Keywords: mouse study
Funders Acknowledgements:
ASAP
Grant ID: ASAP-000420
Abstract
Mouse studies
Troubleshooting
All mouse experiments were performed in accordance with the European Community Council Directive 2010/63/EU.
In compliance with Italian law (D. Lgs. n. 26/2014), the protocol was approved by the General Direction of Animal Health and Veterinary Drugs of the Italian Ministry of Health (authorization number 483/2023-PR).
C57BL/6n-Atm1Brd Pitrm1+/− mice were obtained from the Sanger Institute (http://www.informatics.jax.org/allele/MGI:5085349).
Animals were housed in groups of three per cage at 22 ± 2 °C, with a 12-hour light–dark cycle and 60% relative humidity.
Food (rodent standard diet; catalog number 4RF25, Mucedola, Settimo Milanese, Milan, Italy) and water were provided ad libitum.
The experimental design included two groups of female mice, aged 6 months and 15 months, consisting of 6 Pitrm1+/+ mice and 6 Pitrm1+/− mice.
For histological and immunohistochemical analyses, brains were fixed in formalin.
One hemisphere of each brain was sectioned and fresh-frozen for Western blot analysis.
For immunohistochemistry, the other hemisphere was frozen and embedded in optimal cutting temperature (OCT) compound (Tissue-Tek, Sakura Finetek, Torrance, CA, USA).
Brain slices of 20 µm thickness were obtained by cryosectioning.
Free-floating sections were stored in antifreeze solution.