Female Sprague-Dawley rats (6–8 weeks of age, weighing 200–250 g) were obtained from the Experimental Animal Center of Shanghai Jiaotong University (Shanghai, China). The ethics committees from Qingdao University Affiliated Hospital have approved this animal study, and the approval was obtained prior to the commencement of the study. All animal studies were carried out following the guidelines and regulations of the Animal Care and Use Committees of Qingdao University Affiliated Hospital. All rats were housed in microisolator cages under specific pathogen-free conditions on a 12 h light–dark cycle. Rat BCa models were constructed by intravesical instillation of N-methyl-N-nitrosourea (MNU, Sigma-Aldrich) at 2 mg/rat every other week for a total of four doses. At the end of the 8th week, all rats received abdominal computed tomography examination to observe the tumorigenic situation. Three rats were sacrificed and their bladders were harvested for histological study.
N methyl n nitrosourea mnu
Manufactured by Merck Group
Sourced in United States
N-methyl-N-nitrosourea (MNU) is a chemical compound used in laboratory research. It serves as a DNA alkylating agent, which can induce genetic mutations. As a synthetic compound, MNU has applications in various experimental settings, but its specific intended uses are not provided here.
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10 protocols using n methyl n nitrosourea mnu
1
Rat Bladder Cancer Induction Model
2
Photoreceptor Degeneration Induction
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Photoreceptor degeneration was induced by a single intraperitoneal injection of N-methyl-N-nitrosourea (MNU, Sigma-Aldrich, St. Louis, MO, USA) at the dose of 60 mg/kg body weight for mice (3 weeks old) and 70 mg/kg body weight for rats (5 weeks old), as reported previously5 (link).
3
MNU-Induced Tumor Model in Transgenic Mice
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Male C57BL/6N mice were purchased from Charles River (Hino, Japan). Starting at 3 weeks of age, Rev1 Tg and normal C57BL/6N male mice were treated with 25 mM ZnSO4 in drinking water to induce transgenic Rev1 expression. Beginning at 6 weeks of age, the mice were injected intraperitoneally with N-methyl-N-nitrosourea (MNU) (SIGMA, Japan), 50 mg/kg of body weight, once per week for 2 weeks. Mice were observed daily until becoming moribund, and then sacrificed under anesthesia and necropsied. Concentration of hemoglobin (Hgb) in peripheral blood was measured using a PCE-310 hygrometer (ERMA). Tissue was collected from MNU-treated animals for weight measurements and histology.
4
Cytotoxicity Assay Using MNU, Cisplatin, and 5-FU
5
Modulating Retinal Degeneration in Zebrafish
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The fish were randomly assigned to either the uninhibited or the inhibited group. In the latter group, the TGFβ/activin pathway was blocked using the small molecule inhibitor SB431542 (Tocris, Bristol, UK). The inhibitor was dissolved in dimethyl sulfoxide (DMSO) and added to the water of the fish tank, beginning one day prior to the induction of retinal degeneration, and was refreshed every third day. The final concentration in the water of the fish tank was 20 μM SB431542 and 0.1% DMSO. The uninhibited fish were held in water with 0.1% DMSO. Retinal degeneration was induced in both groups by placing the zebrafish in water containing 150 mg/l N-methyl-N-nitrosourea (MNU, Sigma, St. Louis, MO, USA) for one hour as previously described by our group [14 (link)].
6
Gastric Cancer Mouse Model Development
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All protocols involving animal models or animal specimens were approved by the Ethical Committee of Xi'an Jiaotong University Medical College and the Experimental Animal Center of Xi'an Jiaotong University. The experimental schematic is shown in Fig. 1 D. Forty C57BL/6 male mice were divided into two groups. N-Methyl-N-nitrosourea (MNU, Sigma, St. Louis, MO, USA) was dissolved in dH2O at a concentration of 30 p.p.m., and this solution was freshly prepared as drinking water twice per week. The drinking water was changed to normal dH2O on alternating weeks for 14 weeks, giving 7 total weeks of MNU exposure. Normal dH2O was used as the drinking water for the control group. All mice were euthanized by phenobarbitone (0.3 mg/10g body weight) at 50 weeks from the beginning of MNU administration. Gastric tissues were collected as previously described.
7
Photoreceptor Degeneration Induction
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To induce photoreceptor degeneration, N-methyl-N–nitrosourea (MNU, Sigma-Aldrich, St. Louis, MO, USA) was intraperitoneally injected into rats at the dose of 70 mg/kg body weight as described previously5 (link).
8
Inducing Photoreceptor Degeneration with MNU
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Photoreceptor degeneration was induced by a single intraperitoneal injection of N-methyl-N-nitrosourea (MNU, Sigma-Aldrich, St. Louis, MO, USA, 70 mg/kg body weight) as reported previously16 (link).
9
Tadalafil Characterization and MNU Procurement
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Tadalafil (API) was procured from the Sanofi Aventis as a gift sample followed by FTIR analysis to establish its purity. N-methyl-n-nitrosourea (MNU) (Sigma-Aldrich, N1517) was procured from Sigma Life Science Aldrich Co. 3050 Spruce Street, St. Louis, USA. Other chemicals were purchased from Himedia Pvt. Ltd., Sigma Aldrich and Amresco.
10
Synthesis and Procurement of Carcinogenic Compounds
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PhIP was synthesized in the NARD Institute (Osaka, Japan), with a purity of 99.9%. N-nitrosobis(2oxopropyl)amine (BOP) was obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). 7,12dimethylbenz[a]anthracene (DMBA) and N-methyl-Nnitrosourea (MNU) were obtained from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). N-methyl-N'-nitro-Nnitrosoguanidine (MNNG), dimethylnitrosamine (DMN) and 1,2-dimethylhydrazine (DMH) were obtained from Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan). 3,2'dimethyl-4-aminobiphenyl (DMAB) was obtained from Matsugaki Pharmaceutical Co. (Osaka, Japan).
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