Male C57Bl/6 mice (Taconic Labs) were injected intraperitoneally (i/p) with 55 mg/kg of STZ (Sigma Aldrich, St. Louis, MO) diluted in sterile citrate buffer (0.05 M, pH 4.5) were administered daily for four consecutive days. Control mice received i/p injection of citrate buffer (0.05 M, pH 4.5) alone. At 26 weeks after injection of STZ or citrate buffer all animals were tested for blood glucose level through tail vein puncture and using AlphaTRAK2 (Abbott Diabetes Care Inc., Alameda, CA). Animals with blood glucose levels more than 350 mg/dL were considered diabetic. All experimental protocols were approved by the Animal Care and Use Committee of Xavier University of Louisiana and were conducted in accordance with the ARVO statement for the use of Animals in Ophthalmic and Vision Research.
Alphatrak2
Manufactured by Abbott
Sourced in United States, Israel
The AlphaTrak2 is a blood glucose monitoring system designed for veterinary use. It provides a simple and convenient way to measure blood glucose levels in pets. The device utilizes advanced technology to deliver accurate and reliable results, enabling veterinary professionals to effectively monitor and manage their patients' blood glucose levels.
Automatically generated - may contain errors
Lab products found in correlation
Alphatrak2 test 2 strips, by Abbott (2 mentions)
Humulin r, by Eli Lilly (2 mentions)
Streptozotocin (stz), by Abbott (1 mentions)
Streptozotocin (stz), by Merck Group (1 mentions)
Tyloxapol, by Merck Group (1 mentions)
Free glycerol reagent, by Merck Group (1 mentions)
Novolin, by Novo Nordisk (1 mentions)
Ab10983, by Abcam (1 mentions)
Edta coated microvette tubes, by Sarstedt (1 mentions)
Vetsulin, by Merck Group (1 mentions)
Meso scale discovery two plex mouse metabolic immunoassay kit, by Mesoscale (1 mentions)
23 protocols using alphatrak2
1
Streptozotocin-Induced Diabetic Murine Model
2
Daily Whole Blood Glucose Monitoring
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Whole blood glucose levels were measured daily using an AlphaTRAK2 glucose metre (Abbott Diabetes Care Inc., Alameda, CA, USA) from a sterile saphenous vein bleed. Blood glucose values reported here were recorded just prior to skeletal muscle harvest.
3
Canine Blood Glucose Screening
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Owners of eligible dogs were asked to attend with their dog for a screening appointment, at which signed consent to participate was obtained. Jugular vein blood samples were collected and submitted for biochemistry and haematology, and whole blood glucose measurement was performed using a handheld device (Alphatrak 2; Abbott Diabetes Care Inc., CA, USA) validated for measurement of glucose concentrations in canine blood.
4
Assessing Metabolic Physiology in Mice
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TG clearance was assessed in mice fasted overnight (1700–0900 h) after an oral gavage of 200 μL of food-grade olive oil (Oleoestepa, Estepa, Spain). The rate of hepatic VLDL-TG secretion was assessed after tyloxapol injection (500 mg/kg i.p.; Sigma-Aldrich). Glucose tolerance tests were performed in mice fasted overnight that were injected with 2 mg glucose/g BW i.p. Insulin tolerance tests were performed in mice 4 h after food removal, starting at 0700 h by injecting 1.5 mU insulin/g BW i.p. (Novolin; Novo Nordisk, Bagsvaerd, Denmark). Blood samples were taken from lateral tail vein, and blood glucose measured (AlphaTRAK 2; Abbott Laboratories, Abbott Park, IL). To assess ex vivo WAT lipolysis, 50–60 mg of tissue from urogenital fat pads was washed in cold PBS and minced into small pieces, then incubated in 500 μL Krebs-Ringer HEPES buffer without or with 1 µmol/L isoproterenol for 2 h at 37°C 5% CO2. Media were collected to measure glycerol production (Free Glycerol Reagent; Sigma-Aldrich). Hepatic insulin sensitivity was assessed in mice fasted overnight that were injected with saline or 2 mU/g BW insulin i.p. Twenty minutes later, the liver was collected for Western blot analysis.
5
Metformin Effects on Insulin Sensitivity in Mice
6
Glyburide Dosage and Blood Glucose Measurement
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For glyburide dosing we obtained commercially available generic glyburide tablets of 1.5 mg (Teva) and 1.25 mg (Heritage) each. Blood glucose was measured using a commercially-available glucometer and test strips designed for diabetic monitoring in dogs (AlphaTrak2; Abbott Laboratories, Abbott Park, IL, USA); the performance of this testing method against reference standards has been previously published (Cohen et al., 2009 (link)).
For measurement of glyburide concentration in blood samples, control normal canine plasma was obtained from Equitech-Bio, Inc. (#SCAPE35-0100; Kerrville, TX, USA). Glyburide (USP Reference, #1295505) and Glipizide (G117) reference materials were obtained from Sigma (St. Louis, MO, USA). PBS Buffer, pH 7.0, was made using 8 g NaCl, 0.2 g KCl, 1.44 g Na2HPO4, and 0.24 g KH2PO4 dissolved in 1 L reverse-osmosis deionized (RO-DI) water, pH adjusted to 7.0 with a 6N HCl solution, sourced in-house. All chemicals and reagents were ACS grade and obtained from VWR Scientific (Randor, PA, USA).
For measurement of glyburide concentration in blood samples, control normal canine plasma was obtained from Equitech-Bio, Inc. (#SCAPE35-0100; Kerrville, TX, USA). Glyburide (USP Reference, #1295505) and Glipizide (G117) reference materials were obtained from Sigma (St. Louis, MO, USA). PBS Buffer, pH 7.0, was made using 8 g NaCl, 0.2 g KCl, 1.44 g Na2HPO4, and 0.24 g KH2PO4 dissolved in 1 L reverse-osmosis deionized (RO-DI) water, pH adjusted to 7.0 with a 6N HCl solution, sourced in-house. All chemicals and reagents were ACS grade and obtained from VWR Scientific (Randor, PA, USA).
7
Glucose Tolerance Test in Mice
8
Streptozotocin-Induced Diabetic Model
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All animals were made diabetic using a single intraperitoneal injection of streptozotocin (STZ) dissolved in saline (60 mg/kg; Sigma‐Aldrich, St. Louis, MO, USA). A 10% sucrose solution was provided in place of the drinking water for the first 24 hours after STZ administration to prevent hypoglycaemia. After that, the animals received regular drinking water. Diabetes was maintained for a period of 2 weeks, and during this time, blood glucose was monitored twice daily with the AlphaTRAK2 glucometer (Abbott Laboratories; Chicago, IL, USA), and a variable subcutaneous dose of protamine zinc insulin (Boehringer Ingelheim; Duluth, GA, USA) was administered once per day to keep morning glucose levels below 500 mg/dL.
9
Recurrent Hypoglycemia Induction Protocol
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Recurrent hypoglycaemia was initiated 3 days before the terminal clamp procedure or isoproterenol test. Hypoglycaemia was induced using a single intraperitoneal injection of regular insulin (30–40 IU/kg, Humulin R, Eli Lilly, Indianapolis, IN, USA). Plasma glucose concentrations were monitored through a tail nick every 30 minutes using an AlphaTRAK 2 glucometer (Abbott Laboratories, Chicago, IL, USA), and glucose levels were kept between 30 and 40 mg/dL for at least 1.5 hours (Supplemental Table 1). During recurrent hypoglycaemia, animals had free access to water, but not food. Following each episode of hypoglycaemia, plasma glucose levels were recovered to euglycemic levels by providing free access to food and water. In some cases, 0.5–1 mL of 20% dextrose was injected intraperitoneally to aid recovery if the animals did not eat on their own. Once recovered, the animals were returned to their home cages. This procedure was repeated once daily for 3 consecutive days. Control animals received a saline injection and underwent the same monitoring procedures.
10
Canine Glucose, Cytokine, and Adipose Analysis
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Blood glucose concentrations were measured with a hand-held point-of-care glucose monitor that was previously validated for use in dogs (AlphaTRAK 2; Abbott Animal Health, Abbott Park, IL, USA). Blood samples for cytokine and hormone analysis were collected into chilled EDTA tubes and plasma was separated within 2 h.
Plasma was stored in −20° C and was analyzed in one batch at the end of the study. The analysis was performed in blind fashion. Plasma levels of canine IL6, MCP1, GMCSF, TNFα, insulin, and leptin were measured with MILLIPLEX MAP Canine Cytokine/Chemokine and Canine Endocrine Magnetic Bead Panels, Millipore/Merck, Darmstadt, Germany, Cat No CCYTOMAG-98K). All samples were tested in duplicates.
Paraffin embedded sections of right and left side of injected falciform fat were used for hematoxylin and eosin staining (H&E), as well as immunohistochemistry analysis using rabbit polyclonal antibodies against mouse UCP1 (Abcam, ab10983, react with canine and mouse UCP1) was performed as previously descriebed15 (link). Antibody was used at 1:1000 dilution.
Plasma was stored in −20° C and was analyzed in one batch at the end of the study. The analysis was performed in blind fashion. Plasma levels of canine IL6, MCP1, GMCSF, TNFα, insulin, and leptin were measured with MILLIPLEX MAP Canine Cytokine/Chemokine and Canine Endocrine Magnetic Bead Panels, Millipore/Merck, Darmstadt, Germany, Cat No CCYTOMAG-98K). All samples were tested in duplicates.
Paraffin embedded sections of right and left side of injected falciform fat were used for hematoxylin and eosin staining (H&E), as well as immunohistochemistry analysis using rabbit polyclonal antibodies against mouse UCP1 (Abcam, ab10983, react with canine and mouse UCP1) was performed as previously descriebed15 (link). Antibody was used at 1:1000 dilution.
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