Both male and female C57BL/6 (wild type; WT), GFP reporter under the insulin promoter on C57BL/6 background (INS-GFP), and NOD.CB17-Prkdcscid (NOD.SCID) mice at 8–12 weeks of age were used as a source for pancreatic islets. NOD/ShiLtj (non-obese diabetic; NOD)29 (link),30 (link) and NOD.CB17-Prkdcscid (NOD.SCID) female mice at 16–20 weeks of age served as recipients of intraperitoneal (i.p) implantations with islet-seeded biomaterial, islets-only or biomaterial-only. NOD diabetic mice received syngeneic pancreatic islets derived from NOD.CB17-Prkdcscid donor mice. For streptozotocin (STZ)-induced diabetes, low dose STZ (Cayman Chemicals; Ann Arbor, MI) (12 g/kg) treatment was i.p. injected each day over the course of five days into both male and female C57BL/6 WT mice at ages 10–16 weeks old31 (link),32 (link). STZ-induced diabetic mice received syngeneic pancreatic islets from healthy (i.e. non-STZ treated) donor WT mice. Littermates were used for age- and sex-matching. Animal procedures were performed in accordance and approved by the Institutional Animal Care and Use Committee. Mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and housed in pathogen-free facilities at Howard University.
Streptozotocin (stz)
Manufactured by Cayman Chemical
Sourced in United States
STZ is a laboratory reagent used for biomedical research. It is a chemical compound that serves as a core functional component in various experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Antibody against β actin, by Abcam (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Am251, by Cayman Chemical (1 mentions)
Antibody against bax, by Santa Cruz Biotechnology (1 mentions)
Win55 212 2, by Cayman Chemical (1 mentions)
Formaldehyde solution, by Avantor (1 mentions)
Poloxamer 188 10 solution, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
H e stain, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Dulbecco modified eagle medium (dmem), by GE Healthcare (1 mentions)
Carbopol 940, by Thermo Fisher Scientific (1 mentions)
Miglyol 812, by Sasol (1 mentions)
Precirol ato 5, by Gattefosse (1 mentions)
Metformin hydrochloride, by Cayman Chemical (1 mentions)
Nicotinamide, by Cayman Chemical (1 mentions)
Arachidonyl 2 chloroethylamide, by Cayman Chemical (1 mentions)
Jzl195, by Cayman Chemical (1 mentions)
22 protocols using streptozotocin (stz)
1
Intraperitoneal Implantation of Pancreatic Islets
2
Genetically Modified Mouse Models for Diabetes
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Mouse models included Clec16aloxP (13 (link)), Ins1-Cre (55 (link)), mt-Keima (ref. 19 (link) and a gift from Toren Finkel, University of Pittsburgh, Pittsburgh, Pennsylvania, USA), and NOS2–/– mice (56 (link)). mt-Keima mice were separately maintained on the FVB/N and C57BL/6N (B6N) backgrounds, while all other models were maintained on the B6N background. Clec16aloxP were mated to Ins1-Cre mice, and B6N mt-Keima were mated to NOS2–/– mice to generate experimental groups. For studies using the conditional Clec16a allele, Ins1-Cre–alone mice and Clec16aloxP/loxP mice were phenotypically indistinguishable from each other and, thus, combined as WT controls. For STZ studies, 5-week-old male mice were injected with 50 mg/kg STZ (Cayman Chemical) for 5 consecutive days (days 1–5). Randomly fed blood glucose concentrations were measured 3–4 times weekly for up to 35 days after STZ. Animals were housed on a standard 12-hour light/12-hour dark cycle with ad libitum access to food and water.
3
Streptozotocin-induced Diabetes in Mice
4
Molecular Mechanisms of Cannabinoid Signaling
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AM251(N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide),WIN55,212-2([(3R)-2,3-dihydro-5-methyl-3-(4 morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenyl-methanone) and streptozotocin (STZ) (2-deoxy-2-[[(methylnitrosoamino)carbonyl] amino]-D-glucose) were purchased from Cayman Chemical. Rimonabant (biaryl pyrazole N-(piperidinyl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide) was synthesized at Ajou University. Antibodies against caspase-3, cyclin D2, and Bcl-2 were purchased from Cell Signaling Technology. An antibody against β-actin was purchased from Abcam. Antibody against Bax was purchased from Santa Cruz. Anti-mouse and anti-rabbit secondary horseradish peroxidase conjugate (HRP) was obtained from Bio-Rad Laboratories.
5
Streptozotocin-Induced Type 2 Diabetes in Rats
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Streptozotocin (STZ; Cayman Chemical, Ann Arbor, MI, USA) was administered intraperitoneally (i.p.) at a dose of 35 mg/kg to rats that reached the target weight for obesity to create a DM-2 model [18 (link)]. Two days after STZ administration, only 20 of 32 rats could be included in the study, since their glucose levels were higher than 200 mg/dL according to the results of blood samples taken from the tail vein. STZ was readministrated to these 12 rats. Two days later, 7 of these STZ-treated rats had glucose levels of 200 mg/dL or above and were also included in the study. A total of 27 obese and diabetic rats were thus included in the study. DM-2 was controlled with glucose measurements at 15-day intervals.
6
Streptozotocin Acquisition for Research
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The chemical streptozotocin (STZ) was purchased from Cayman Chemical (Ann Arbor, MI, USA).
7
Analgesic Drugs and Enzyme Inhibitor Evaluation
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Drugs were purchased from different companies: deltorphin II was from AnaSpec, SNC‐80 ((+)‐4‐[(αR)‐α‐((2S,5R)‐4‐allyl‐2,5‐dimethyl‐1‐piperazinyl)‐3‐methoxybenzyl]‐N,N‐diethylbenzamide) was from Tocris Cookson, SB235863 ([8R‐(4bS*,8aα,8aβ,12bβ)]7,10‐dimethyl‐1‐methoxy‐11‐(2‐ethylpropyl)oxycarbonyl 5,6,7,8,12,12b‐hexahydro‐(9H)‐4,8‐methanobenzofuro[3,2‐e]pyrrolo[2,3‐g]isoquinoline hydrochloride) was a generous gift from Dr L Gendron17 (University of Sherbrooke, QC, Canada), and TIPP was from Cedarlane. Streptozotocin (STZ) was purchased from Cayman Chemical. The endothelin‐converting enzyme 2 (ECE2) inhibitor, 6634449, was purchased from Vitas‐M laboratory (product code STK521587).
8
Glyceryl Palmitostearate Formulation for Therapeutic Delivery
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Precirol ATO5 (glyceryl palmitostearate) was obtained from Gattefosse (Saint-Priest, France) and Miglyol 812 was received from Sasol (Hamburg, Germany). Carbopol 940 was purchased from Acros (Morris County, NJ, USA). Triethanolamine (TEA) was purchased from Merck (Darmstadt, Germany). RhTM protein and anti-rhTM antibody were provided by Blue Blood Biotech Corporation (Taipei, Taiwan). H&E stain, Poloxamer 188 (10% solution), bovine serum albumin (BSA) and phosphate buffered saline (PBS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Streptozotocin (STZ) was obtained from Cayman Chemical (Ann Arbor, MI, USA). DMEM and fetal bovine serum (FBS) were purchased from GE Healthcare (Pittsburgh, PA, USA). Formaldehyde solution was from Avantor (Center Valley, PA, USA), and penicillin-streptomycin was from Gibco (Waltham, MA, USA).
9
Pharmacological Agents in Biochemical Assays
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Fine chemicals such as streptozotocin, metformin hydrochloride, and nicotinamide were procured from Cayman Chemical, USA. ELISA kits for estimation of insulin, acetylcholine, acetylcholinesterase were procured from Cloud-Clone Corp., USA. Other chemicals and solvents were purchased locally with analytical grades.
10
Streptozotocin-Induced Diabetic Rat Model
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The animals were divided into 6 groups with 5–8 animals each (Figure 1 ), and then fasted for 12 h and subsequently submitted to diabetes mellitus induction by a single intraperitoneal injection of streptozotocin (STZ) of 45 mg/kg (Cayman Chemicals, Ann Arbor, USA) dissolved in pH 4.5 citrate buffer [35 (link)].
In the first 24 h after induction, animals received 10% glucose solution to prevent hypoglycemia. Diabetes was confirmed by the elevated glucose levels in plasma, determined at 72 h. The animals with blood glucose greater than or equal to 250 mg/dL or 15 mM [36 (link)] were considered diabetic.
At the end of the experimental period (4 weeks), the animals were euthanized with an overdose of sodium thiopental (150 mg/kg, IP) preceded by lidocaine 10 mg/kg, IP. Immediately after euthanasia, blood and liver samples were collected for further analysis.
The doses of BSB used were based on previously performed studies on Platonia insignis Mart. biological effects [17 (link)]. The recommended insulin dose was used following the proposed by previous studies with similar DM model [37 (link)].
In the first 24 h after induction, animals received 10% glucose solution to prevent hypoglycemia. Diabetes was confirmed by the elevated glucose levels in plasma, determined at 72 h. The animals with blood glucose greater than or equal to 250 mg/dL or 15 mM [36 (link)] were considered diabetic.
At the end of the experimental period (4 weeks), the animals were euthanized with an overdose of sodium thiopental (150 mg/kg, IP) preceded by lidocaine 10 mg/kg, IP. Immediately after euthanasia, blood and liver samples were collected for further analysis.
The doses of BSB used were based on previously performed studies on Platonia insignis Mart. biological effects [17 (link)]. The recommended insulin dose was used following the proposed by previous studies with similar DM model [37 (link)].
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