We used 10-week-old adult female C57BL/6 mice and 10-week-old adult female Thy1-GFP-M mice (Jackson Laboratory, USA). All the animals were divided randomly into the following three groups (n=12 animals per group): 1) animals that received injections of a vehicle solution (4% PEG-400, 2% DMSO and 94% saline) beginning 1 h after injury and continuing on a daily basis for 7 d (Vehicle control group), 2) animals that received T10 laminectomy without SCI and injections of a vehicle solution (4% PEG-400, 2% DMSO and 94% saline) beginning 1 h after injury and continuing on a daily basis for 7 d (Sham-operated control group), and 3) animals that received injections of FFA dissolved into the vehicle solution as previously described (Treatment group). All animal experiments were performed in strict accordance with the Guide for the Care and Use of Laboratory Animals (National Research Council, 1996), and all efforts were made to minimize suffering. All animal procedures were approved by the Hubei Provincial Animal Care and Use Committee and complied with the experimental guidelines of the Animal Experimentation Ethics Committee of Huazhong University of Science and Technology in China.
Thy1 gfp m mice
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Thy1-GFP-M mice are a transgenic mouse line that expresses green fluorescent protein (GFP) under the control of the Thy1 promoter. This results in the labeling of a subset of neurons in the mouse brain with GFP, allowing for visualization and study of these cells.
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5 protocols using thy1 gfp m mice
1
Evaluating Therapeutic Efficacy of FFA in SCI Mice
2
Thy1-GFP-M Mouse Protocol
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In this study, adult (two- to three-month-old) Thy1-GFP-M mice (Jackson Laboratory) were used. All experimental procedures were performed in strict accordance with the Experimental Animal Management Ordinance of Hubei Province, China, and were approved by the Institutional Animal Ethics Committee of Huazhong University of Science and Technology.
3
Genetic Manipulation of PSD-Zip70 in Mice
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PSD-Zip70KO mice (RRID: MGI:5706982 ) (Mayanagi et al., 2015 (link)) and the parental wild-type (WT) strain C57BL/6J (Japan SLC) were used as subject mice. For morphological analyses, PSD-Zip70KO mice were crossed with Thy1-GFP-M mice (JAX Mice, The Jackson Laboratory) (RRID:IMSR_JAX:007788 ) (Feng et al., 2000 (link)). Thy1-GFP-M mice were the same genetic background (C57BL/6J) as PSD-Zip70KO mice. After receipt of mating pairs from The Jackson Laboratory, Thy1-GFP-M mice were additionally backcrossed to C57BL/6J for 3 generations before use. We used The GFP-positive PSD-Zip70KO (-/-) homozygous and WT [PSD-Zip70 (+ / +)] offspring were used. Animals were housed in groups with a 12-h light/dark cycle (lights on 7:00 to 19:00) in the Center for in vivo Sciences of Iwate Medical University. All of the procedures involving animals and their care were approved by the Animal Care Committees of the authors’ institutes, and were carried out according to their guidelines for animal experiments.
4
In Vivo Imaging of Neuronal Activity
5
Experimental Mice for Neuroscience Studies
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Male mice, aged 2–4 months, were used for all experiments. For experiments in wild-type animals, C57BL/6J mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). For in vivo brain microdialysis experiments, C57BL/6NCrl mice (Martinsried, Germany) as well as global Fkbp5−/−58 (link) mice and respective wild type controls (Martinsried, Germany) were used. Neonatal (P7-9) Thy1-GFP M mice (sex not determined) with a sparse expression of green fluorescent protein (GFP) in principal neurons in cortex and hippocampus, Jackson Laboratory Stock #007788)59 (link) were used in organotypic hippocampal slice cultures (OHSC) experiments. All animals were kept under standard laboratory conditions and were maintained on a 12 h light–dark cycle (lights on from 0700 h to 1900 h), temperature 22 ± 1 °C, humidity 50%, in clear Plexiglas cages (19 × 29 × 13 cm3) with unrestricted access to food (Purina Laboratory Rodent Diet 5001) and water.
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