Adult (2- to 3-month-old) male and female C57BL6 mice, Cdh5BAC-GCaMP8 mice, and NG2-DsRed mice (C57/BL6J background; Jackson Laboratories) were group-housed on a 12-hour light:dark cycle with environmental enrichment and free access to food and water. All animal procedures received prior approval from the University of Vermont Institutional Animal Care and Use Committee.
Ng2 dsred mice
Manufactured by Jackson ImmunoResearch
NG2-DsRed mice are a transgenic mouse line that express the red fluorescent protein DsRed under the control of the NG2 chondroitin sulfate proteoglycan promoter. NG2 is a marker for oligodendrocyte precursor cells (OPCs) and pericytes. The DsRed fluorescent protein allows for the visualization and identification of NG2-expressing cells in this mouse model.
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4 protocols using ng2 dsred mice
1
Longitudinal Calcium Imaging in Mice
2
Genetically Engineered Mice Imaging
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NG2-dsRed mice (stock 008241) were purchased from The Jackson Laboratory. LifeAct–EGFP mice were generated previously [31 (link)], and provided by R. Wedlich-Söldner (Max-Planck Institute of Biochemistry, Martinsried, Germany) and L. M. Machesky (Beatson Institute for Cancer Research, Glasgow, UK). H2B-EGFP mice (stock 006069) were purchased from The Jackson Laboratory.
Approximately 30 mice were used to perform the described experiments. Mice were housed under the approval and the institutional guidelines governing the care of laboratory mice of the Italian Ministry of Health, under authorization number 1073/2015-pr and in compliance with the international laws and policies.
Approximately 30 mice were used to perform the described experiments. Mice were housed under the approval and the institutional guidelines governing the care of laboratory mice of the Italian Ministry of Health, under authorization number 1073/2015-pr and in compliance with the international laws and policies.
3
Transgenic Mouse Models for Imaging
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Homozygous transgenic Flk1-myr::mCherry and Csf1r-EGFP mice were generated as described previously [42 (link)–44 (link)]. NG2-DsRed mice were purchased from The Jackson Laboratory. This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All animal research was conducted according to protocols approved by the Institutional Animal Care and Use Committee (IACUC) of Baylor College of Medicine (assurance number AN-4593).
4
Streptozotocin-Induced Diabetic Mouse Model
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Diabetes was induced in three mouse lines: C57BL/6 mice (Jackson Laboratory, Stock#: 000664, RRID:IMSR_JAX:000664), NG2-DsRed mice (Jackson Laboratory, Tg(Cspg4-DsRed.T1)1Akik/J, Stock#: 008241, RRID:IMSR_JAX:008241) and NG2-Cre-GCamp6f, generated by crossing FVB-lfi208Tg(Cspg4-cre)1Akik/J (Jackson Laboratory, Stock#: 008533, RRID:IMSR_JAX 008533) with Ai95(RCL-GCaMP6f)-D loxP (Jackson Laboratory, Stock#: 024195, RRID:IMSR_JAX 024195). Breeding pairs negative for the rd1 mutation were used to produce animals for this study. We used the STZ diabetic mouse model57 (link). Male mice aged 6–8 weeks were fasted for 4 h prior to the injections. The animals were injected intraperitoneally on 5 consecutive days with 50 mg/kg STZ (Sigma-Aldrich, S0130) freshly dissolved in a citrate buffer (pH 4.5). Control animals received a citrate buffer injection without STZ. In our STZ mouse model of diabetes, the levels of blood glucose reached maximum elevation 1 month after STZ injection and remained elevated. The diabetes was defined by non-fasting blood glucose >300 mg/dL verified 1 month after the last STZ injection and confirmed on the day of the experiment.
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