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Aldh1l1 cre ert2 mice

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The Aldh1l1-Cre/ERT2 mice are a transgenic mouse line that expresses a tamoxifen-inducible Cre recombinase under the control of the Aldh1l1 (Aldehyde Dehydrogenase 1 Family Member L1) promoter. The Aldh1l1 gene is expressed in astrocytes, a type of glial cell in the central nervous system. The Cre/ERT2 fusion protein allows for temporal and spatial control of gene recombination in astrocytes upon administration of tamoxifen.

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4 protocols using aldh1l1 cre ert2 mice

1

Genetic Labeling and Manipulation of Astrocytes in Mice

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Adult male and female mice and postnatal pups were used on a C57Bl/6J background (#000664, The Jackson Laboratory). B6.Cg-Tg(Gfap-cre)73.12Mvs/J mice52 (link) (The Jackson Laboratory, #012886) mice were crossed with B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato) Hze/J mice53 (link) (The Jackson Laboratory, #007909) to generate Gfapcre/+TdTomatof/+ (TdTomatoGfap) mice. B6N.129-Rpl22tm1.1Psam/J mice54 (link) (The Jackson Laboratory, #011029) were bred to Gfapcre mice to generate Gfapcre/+Ribotagf/f (RibotagGfap) mice. Aldh1l1creERT2 mice55 (link) (The Jackson Laboratory, #029655) were bred to Csf2rbf/f mice39 (link) to generate Aldh1l1creERT2Csf2rbf/f mice. Conditional deletion of Csf2rb was induced at 5 weeks of age with tamoxifen (225 mg/kg; Sigma-Aldrich, #T5648) diluted in corn oil (Sigma-Aldrich, #C8267); EAE was induced 3 weeks later. Mice were kept in a pathogen-free facility at the Hale Building for Transformative Medicine at Brigham and Women’s Hospital in accordance with the IACUC guidelines. Eight-to-twelve-week-old mice were used for stereotactic injection and EAE induction. Pups were killed between postnatal day 0 (P0) and P3 for collection and culture of astrocytes. All procedures were reviewed and approved under the IACUC guidelines at Brigham and Women’s Hospital.
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2

Characterization of Mouse Neural Populations

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All experiments were approved by the Vanderbilt University Institutional Animal Care and Use Committees and were conducted in accordance with the National Institute of Health guidelines for the Care and Use of Laboratory Animals. 5–16 week-old male and female C57BL/6J mice obtained from Jackson Labs or bred in-house and were used for experiments throughout the manuscript. 10–18 week-old male and female Cnr1flx/flx mice bred in-house were used for experiments in Figure 3. Aldh1l1-Cre/ERT2 mice and Ai95D mice were obtained from Jackson Labs and crossed in-house to obtain Aldh1l1-Cre:Ai95D mice. 10–18 week-old male and female Aldh1l1-Cre:Ai95D mice were used for experiments in Figure 3. SOM-IRES-Cre mice and Ai14 mice were obtained from Jackson labs and crossed in-house to obtain SOM:Ai14 mice. 10–20 week-old male and female SOM:Ai14 mice were used for experiments in Figure 2. SOM:Ai14 mice were used over SOM-IRES-Cre mice due to the phenotype observed in homozygous SOM-IRES-Cre mice39 (link) and the availability of this line within our laboratory, as the Ai14 reporter construct should not interfere with our experimental strategy. Sex differences were not a primary analysis variable in this study and no overt sex differences were observed, thus all data are pooled from both sexes.
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3

Conditional Xbp1 Deletion in Mice

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All experiments were reviewed and approved by the Brigham & Women’s Hospital Institutional Animal Care and Use Committee. Adult mice aged 8–10 weeks and pups aged from postnatal day (P)0–P3 were used on a C57Bl/6J background (000664, The Jackson Laboratory). Aldh1l1-cre/ERT2 mice43 (link) (The Jackson Laboratory, 029655) were bred to Xbp1fl/fl mice42 (link) (a gift from L. Glimcher) to generate Aldh1l1-creERT2;Xbp1fl/fl mice. Conditional deletion of Xbp1 was induced at 6–10 weeks of age by gavage with 225 mg kg−1 tamoxifen (Sigma-Aldrich, T5648), diluted in corn oil (Sigma-Aldrich, C8267); EAE was induced 4 weeks later.
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4

Generation of 5XFAD APOE4 Transgenic Mice

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All animal studies conducted abided by institutional animal care and use committee (IACUC) protocols approved by the Animal Studies Committee of Washington University in St. Louis. 5XFAD mice (line Tg7031), a gift from R. Vassar at Northwestern University [52 (link)], were crossed for multiple generations to APOE4flox/flox mice [53 ] to generate 5XFAD APOE4flox/flox mice. Aldh1l1-Cre/ERT2 mice (Jackson Laboratories, Stock No. 031008) were also crossed to APOE4flox/flox mice for several generations to generate Aldh1l1-Cre/ERT2 APOE4flox/flox mice. To generate 5XFAD Aldh1l1-Cre/ERT2 APOE4flox/flox mice, 5XFAD APOE4flox/flox males were crossed to Aldh1l1-Cre/ERT2 APOE4flox/flox females. We refer to mice carrying the 5XFAD and Aldh1l1-Cre/ERT2 genes as “5X+AL+” and their littermates without the Aldh1l1-Cre/ERT2 gene as “5X+AL-”. Littermates without the 5XFAD transgene are designated as “5X-” mice. All mice were housed on a normal 12-h light/dark cycle with free access to food and water. There were no sex differences between males and females and the number of mice used can be found in the figure legends.
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