Aldh1l1 creer

Foxo3^fl/fl (Paik et al., 2007 (link)), Nestin‐Cre (Tronche et al., 1999 (link)), Aldh1l1‐CreER (Srinivasan et al., 2016 (link)), and 5xFAD (Oakley et al., 2006 (link)) mouse lines were obtained from the Jackson Laboratories. The aged C57BL/6J mice were obtained from the aging rodent colony of the National Institute on Aging. The sample size was determined based on previous studies (Lian et al., 2016 (link)). Both male and female mice were used, and these are specified in the results and figure legends. Investigators were blinded to the group identities during data collection and analysis. For AAV injections, postnatal day 3 pups were anesthetized via hypothermia and injected i.c.v. free‐hand with 2.5 × 10¹⁰ viral particles per side using a 28‐gauge needle attached to a Hamilton syringe as described previously (Chen et al., 2021 (link); Martini‐Stoica et al., 2018 (link)). RNA sequencing of total RNA was performed using the Illumina platform. Untargeted lipidomics was carried using a Vanquish UPLC and a Lumos orbitrap mass spectrometer (Thermo Fisher Scientific). The detailed methods for these experiments and other standard procedures including qPCR, Western blotting, immunofluorescence staining and associated image acquisition and analysis are provided in Supplementary Information.

All experimental protocols were carried out in compliance with the Guide for the Care and Use of Laboratory Animals (NIH, Bethesda, MD, USA), the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines, and the Thomas Jefferson University Institutional Animal Care and Use Committee (IACUC). Male and female adult mice between 3 and 9 months of age were used for this study. The following transgenic mouse lines were used: Aldh1l1^CreER/+ (The Jackson Laboratory, Stock #029655) and Rosa26^tdTom/tdTom (Ai14) [20 (link),21 (link)]. To identify subpopulations of astrocytes, 12–16 week old Aldh1l1^CreER/+;Rosa26^tdTom/tdTom mice were given 250 mg/kg tamoxifen via oral gavage for 3 consecutive days 2 weeks prior to C2 hemisection (C2Hx) or laminectomy-only surgery.