In this study we used female heterozygous THY-Tau22 mice maintained on a C57Bl6/J background[33 (link)]. THY-Tau22 mice are a well-established model of Tauopathy that express human 4 repeat tau with two frontotemporal dementia-associated point mutations (G272V and P301S) under control of the neuronal driven promoter Thy1.2[33 (link)]. Mice were anesthetized with 4% isoflurane (Vedco, Inc., St. Joseph, MO) and maintained in 3–3.5% isoflurane during all injections. All animals were housed in a temperature and light-cycle controlled facility, and their care was under the guidelines of the National Institutes of Health and an approved IACUC protocol at University of California, Irvine.
Isoflurane
Manufactured by Vedco
Sourced in United States, Macao
Isoflurane is a colorless, volatile liquid anesthetic agent used in veterinary medicine. It is a halogenated ether compound that is administered through inhalation to induce and maintain general anesthesia in animals.
Automatically generated - may contain errors
Lab products found in correlation
Aluminum hydroxide, by Merck Group (8 mentions)
Ovalbumin (ova), by Vedco (6 mentions)
Ova grade 5, by Merck Group (4 mentions)
Ovalbumin (ova), by Merck Group (2 mentions)
Rectal probe, by Physitemp (2 mentions)
Winged butterfly infusion set, by Terumo (2 mentions)
T pump, by Stryker (2 mentions)
Pelco easislicer, by Ted Pella (1 mentions)
Papain, by Merck Group (1 mentions)
Microsprayer aerosolizer, by Penn-Century (1 mentions)
Ethanol c2h5oh, by Merck Group (1 mentions)
Formaldehyde, by Electron Microscopy Sciences (1 mentions)
Leica cm1950, by Leica Biosystems (1 mentions)
Dna lobind tube, by Eppendorf (1 mentions)
Anti ckit apc, by Thermo Fisher Scientific (1 mentions)
Anti cd16 cd32, by BD (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Igg2a antibody, by BioXCell (1 mentions)
Hdm extract, by Greer Laboratories (1 mentions)
Anti cd3 fitc, by BD (1 mentions)
31 protocols using isoflurane
1
Tauopathy Model in Transgenic Mice
2
Murine Model of Allergic Airway Inflammation
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Sensitization and antigen challenge with OVA were performed as previously described.17 (link)18 (link) Mice were immunized by subcutaneously injecting 25 µg of OVA (Grade V; Sigma-Aldrich, St. Louis, MO, USA) absorbed to 1 mg of aluminum hydroxide (Aldrich, Milwaukee, WI, USA) in 200 µL of phosphate-buffered saline (PBS). Subcutaneous injections were administered on days 0, 7, 14, and 21, followed by intranasal OVA challenge (20/50 µL in PBS) performed on days 33 and 35. Subsequently, intranasal OVA challenges were repeated twice per week for 3 months. Age- and gender-matched control mice were treated equally with PBS. All procedures were performed while mice were anesthetized using isoflurane (Vedco, St. Joseph, MO, USA). Mice were sacrificed 24 hours after the final intranasal OVA challenge, and bronchoalveolar lavage (BAL) fluid and lung tissues were obtained for analysis.
All animal procedures were performed in accordance with Laboratory Animal Welfare Act, the Guide for the Care and Use of Laboratory Animals, and the Guidelines and Policies for Rodent Experiments provided by Institutional Animal Care and Use Committee at the School of Medicine, The Catholic University of Korea (apprOVAl number: CUMC-2015-0194-04).
All animal procedures were performed in accordance with Laboratory Animal Welfare Act, the Guide for the Care and Use of Laboratory Animals, and the Guidelines and Policies for Rodent Experiments provided by Institutional Animal Care and Use Committee at the School of Medicine, The Catholic University of Korea (apprOVAl number: CUMC-2015-0194-04).
3
Murine Model of Allergic Asthma
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The mice were immunized by subcutaneously injecting with 25 µg of OVA (grade V; Sigma-Aldrich, St. Louis, MO, USA) adsorbed to 1 mg of aluminum hydroxide (Aldrich, Milwaukee, WI, USA) in 200 µL of phosphate-buffered saline (PBS). The injections were administered on days 0, 7, 14, and 21. Intranasal challenge with 20 µg OVA/50 µL PBS was administered on days 27, 29, and 31 to mice under isoflurane (Vedco, St. Joseph, MO, USA) anesthesia. The intranasal OVA challenges were then repeated twice a week for 3 months. Age- and sex-matched control mice were treated identically but with PBS alone. All of the mice were euthanized 24 hours after the final OVA challenge, at which time bronchoalveolar lavage (BAL) fluid and lung tissues were obtained. All of the animal research procedures were conducted in accordance with the Laboratory Animals Welfare Act, the Guide for the Care and Use of Laboratory Animals, and the Guidelines and Policies for Rodent Experiments mandated by the Institutional Animal Care and Use Committee of the School of Medicine, The Catholic University of Korea.
4
Investigating NAM Supplementation on Immune Response
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Male and female C57BL/6J mice (6–9 weeks old) were fed with a control diet or diet supplemented with NAM (125 mg/day/Kg) (Research Diet Inc, New Brunswick, NJ) for 14 days before immunization. On day 14 mice were anesthetized with isoflurane (Vedco Inc., Saint Joseph, MO) and given 1 μg of S. aureus enterotoxin A diluted in 50 μL of PBS, or PBS alone by intranasal (i.n.) route (Kumar et al., 2013 (link)). Serum was collected 6 h post immunization. Mice continued on the control and NAM supplemented diet for two more days, and then sacrificed. Axillary and brachial lymph nodes (LN) were collected and the isolated LN cells were analyzed by flow cytometry. To measure intracellular IFNγ, cells were restimulated in vitro with 0.1 μg S. aureus enterotoxin A for 4 h and analyzed by flow cytometry.
5
Murine Model of Allergic Airway Inflammation
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The mice were immunized with 25 μg of OVA (chicken egg albumin, grade V, from Sigma-Aldrich, St. Louis, MO, USA) with 1 mg of aluminium hydroxide (alum) in 200 μL of phosphate-buffered saline (PBS). The mice were immunized by subcutaneous injection on days 0, 7, 14, and 21, and then challenged with intranasal OVA (20 μg/50 μL PBS) on day 33, 35, and 37 and then again twice a week for 3 months while anaesthetized with isoflurane (Vedco, St. Joseph, MO, USA). CON mice were treated with PBS in the same manner. Mice were killed 24 hours after the final OVA challenge, and bronchoalveolar lavage fluid (BALF) and lung tissues were obtained.
6
Liver Vasculature Visualization
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Two-month-old mice were euthanized with isoflurane (Vedco Saint Joseph MO). Ink (Hyatt’s) was injected into the common bile duct using a 30-gauge needle. The entire liver was removed, formalin-fixed, and clarified in a 1:2 solution of benzyl alcohol and benzyl benzoate.
7
Immunohistochemistry of CRFOE in Mice
8
Murine Models of Asthma, COPD, and ACO
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Female C57BL/6N mice (Orient, Gyeongi-do, Korea) aged 6 weeks were used in this study. The mice were divided randomly into control, asthma, COPD, ACO-a, and ACO-b groups.
Mice in the asthma group were immunized with 50 μg OVA (chicken egg albumin, grade V; Sigma-Aldrich, St. Louis, MO, USA) in 1 mg aluminum hydroxide (Sigma-Aldrich) in 200 μL phosphate-buffered saline (PBS). Immunization was performed by intraperitoneal injection on days 0, 7, and 14, and intranasal OVA challenges (100 μg/50 μL PBS) were administered on days 21, 22, 23, and 24 under anesthesia with isoflurane (Vedco, St. Joseph, MO, USA). Mice in the COPD group received intratracheally administer PPE (80 U/kg; Elastin Products Company, Owensville, MI, USA) in 100 μL PBS on day 0. Mice in the ACO-a group received the treatments administered to the asthma (OVA) and COPD (PPE) groups. Mice in the ACO-b group were treated intratracheally with 50 μg papain (Sigma-Aldrich) in 100 μL PBS on days 0, 7, 14, and 21. PPE or papain aerosol was created using a Micro Sprayer Aerosolizer (Penn Century Inc., Wyndmoor, PA, USA). The animals were euthanized on day 25 (Fig.1 ).![]()
Mice in the asthma group were immunized with 50 μg OVA (chicken egg albumin, grade V; Sigma-Aldrich, St. Louis, MO, USA) in 1 mg aluminum hydroxide (Sigma-Aldrich) in 200 μL phosphate-buffered saline (PBS). Immunization was performed by intraperitoneal injection on days 0, 7, and 14, and intranasal OVA challenges (100 μg/50 μL PBS) were administered on days 21, 22, 23, and 24 under anesthesia with isoflurane (Vedco, St. Joseph, MO, USA). Mice in the COPD group received intratracheally administer PPE (80 U/kg; Elastin Products Company, Owensville, MI, USA) in 100 μL PBS on day 0. Mice in the ACO-a group received the treatments administered to the asthma (OVA) and COPD (PPE) groups. Mice in the ACO-b group were treated intratracheally with 50 μg papain (Sigma-Aldrich) in 100 μL PBS on days 0, 7, 14, and 21. PPE or papain aerosol was created using a Micro Sprayer Aerosolizer (Penn Century Inc., Wyndmoor, PA, USA). The animals were euthanized on day 25 (Fig.
Pulmonary disease models. ACO, asthma–chronic obstructive pulmonary disease overlap; alum, aluminum hydroxide; i.p., intraperitoneal; i.t., intratracheal; OVA, OVAlbumin; PPE, porcine pancreatic elastase
9
Induction of Transient Colonic Inflammation in Rats
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Transient colonic inflammation was induced by a single intracolonic administration of TNBS solution. The TNBS solution was prepared fresh before the instillation procedure. For 1 ml of the final solution 0.25 ml of TNBS (5% w/v, Sigma) 0.25 ml of water and 0.5 ml of ethanol (C2H5OH, Sigma) were mixed. The final concentration of TNBS was 12.5 mg/ml. Rats were fasted for 24 hours before instillation procedure to provide better access to the colonic lumen. Animals were briefly anesthetized with isoflurane (VEDCO Inc., St.Joseph, MO), a 7–8 cm long catheter made of polyethylene tubing and attached to a 1 cc syringe was inserted into the rat colon for enema administration. After instillation procedure, an animal was held by the tail to avoid any spill of instilled liquid. To assess the severity of developed inflammation, the daily Disease Activity Index (DAI) was calculated followed by MPO assay as previously described [32] (link).
10
Muscle Regeneration and 8-oxoG Role
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For muscle injury, Six-to-eight week male C57BL/6 (WT) mice and Ogg1 KO mice (weight 20–25 g) were anesthetized with inhalation of 1–2% isoflurane (Vedco), and treated with 25 μL of 10 μM (in pyrogen-free saline) cardiotoxin VII4 (CTX, Sigma-Aldrich) via intramuscular injection into the anterior compartments of both hindlimbs. Mice (n = 5 mice/time point) were sacrificed on day 1, 3, 7, 9 and 11 after CTX-induced injury. TA muscles were collected and mRNA levels of MyoD and Myogenin of samples were measured. To detect the role of 8-oxoG in muscle regeneration, after CTX-induced injury, 20 μL of vehicle (1 mM NaOH in saline) and 8-oxoG (100 μM) were injected intramuscularly into the left and right TA of these mice, respectively, on day 1, 2, 3, 5 and 7. TA muscles were collected on day 3 and 7, mRNA levels of MyoD and Myogenin of samples were measured; or in other experiments, TA muscles were collected on day 7, and placed in 10% neutral buffered formalin (NBF) for histological examination to confirm the extent of muscle regeneration, or embedded within Tissue-Tek O.C.T for immunolfluorescence staining of eMyHC-positive regenerated myofibers.
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