Bravo-Sánchez E, Peña-Montes D, Sánchez-Duarte S, Saavedra-Molina A, Sánchez-Duarte E, Montoya-Pérez R. Antioxidants (Basel). J Gastroenterol Hepatol. Ignore the data from this mouse from that point on. The glucose tolerance test is used in clinic and research to identify individuals with im …. Prevention and treatment information (HHS). This procedure is performed to assess pituitary function, adrenal function, insulin sensitivity, and sometimes for other purposes. Ryu Y, Kim YJ, Kim YY, Kim J, Kim SW, Kim H, Ku SY. glucose load (1.5 mg/g) (C and D) was given to 10-week-old fetuin KO and WT mice. Type 2 diabetes is characterized by glucose intolerance, caused by insulin resistance in peripheral metabolic tissues and by impaired glucose-stimulated insulin secretion, the hallmark of beta-cell dysfunction. Blood samples were collected with heparinized capillary tubes from the tail vein before and 30, 60, 90 and 120 min after insulin injection and immediately placed on ice. Before performing the experiment, we will have to determine if the mouse strain is insulin resistant or glucose tolerant, so as to avoid giving the wrong dose of insulin. The intraperitoneal Pyruvate Tolerance Test (iPTT, 1-2g/kg body weight) in 15h-fasted, awake mice is a variant of the intraqperitoneal glucose tolerance test (GTT) in which pyruvate is injected instead of glucose. Dietary advanced glycosylation end products (AGEs) have been linked to insulin resistance in db/db (++) mice. Prepare insulin syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL). Mice were injected with 1.5 mg glucose/g body weight (intraperitoneal). Koren-Iton A, Salomon-Zimri S, Smolar A, Shavit-Stein E, Dori A, Chapman J, Michaelson DM. At approximately 1 min intervals, inject appropriate amount of insulin into interperitoneal cavity of the mouse. Prepare insulin syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL). Normal insulin tolerance and reversal of blood glucose by insulin therapy in Kuma mutant mice. Intraperitoneal Insulin Tolerance Test (Case Western Reserve University) Summary: This is the standard protocol for routine insulin tolerance testing. 2021 Feb 24;10(3):335. doi: 10.3390/antiox10030335. Central and Peripheral Mechanisms in ApoE4-Driven Diabetic Pathology. In vivo metabolic tests are highly valuable to determine whether atherosclerosis progression in mouse models is accompanied by carbohydrate metabolism alterations such as glucose intolerance and insulin resistance. Effects of ambient temperature on glucose tolerance and insulin sensitivity test outcomes in normal and obese C57 male mice 19 May 2015 | Physiological Reports, Vol. 2010 Jul;33(7):e93. For example, BALB/c mice display preserved glucose tolerance and insulin action under HFD, and this seems to be directly related to the lack of fat accumulation, despite lipid accumulation in the muscle. Diabetes Care. This site needs JavaScript to work properly. At the end of the 7th week, an intraperitoneal glucose tolerance test … Glucose tolerance and insulin tolerance tests Animals were fasted overnight and a GTT was performed using glucose (2 g/kg body weight), which was administered by intraperitoneal injection. This protocol details the insulin tolerance test in mice. injection of 0.75 units/kg (E) or 0.15 units/kg (F) regular human insulin. In general for insulin resistant mice, such as those >40g on a high fat diet or such, increase the dose to 2 or 2.5U/kg. At approximately 1 min intervals, inject appropriate amount of insulin into interperitoneal cavity of the mouse. 3, No. In these cases the preferred practice is to measure fat free mass by echoMRI, and then dose animals at approximately 1 U/kg of lean mass for a lean animal or 2.5 U/kg of lean mass for an obese animal. An oral glucose tolerance test (OGTT), in which the mice are challenged with a bolus of glucose and blood glucose and insulin levels are measured across a two hour time course is performed one week prior to the initiation of the HFD challenge. Immobilize mouse and restrain tail with one hand, Aim needle between the midline and the hip bone, Insert syringe (do not inject) into cavity. Fiirst I did glucose tolerance test (2g/kg), fasting overnight (6:pm-9:00 am), after 48 hours, the mice were fasted for 4 hours (8:50 am-1:00 pm), and insulin tolerance test was conducted. 75 units kg −1 body weight under nonfasting conditions. 2-For insulin resistant mice, a higher dose of insulin could be used as they do not respond In general for insulin resistant mice, such as those >40g on a high fat diet or such, increase the dose to 2 or 2.5U/kg. The overall mean at each temperature is also included (pooled). Privacy, Help This test is used to assess the insulin-sensitizing efficacy of test compounds and pharmacological agents that can modify … Insulin Tolerance Test Protocol The electronic version of this document appears in QuaSAR Page 1 of 7 This Document is Uncontrolled in Printed Form Document Code: Insulin_Tolerance_Test_Protocol.doc Controlled Copy ID: Printed on: 22/05/2017 Insulin Tolerance Test Protocol - RNS Endocrinology Test name Insulin tolerance test. For insulin tolerance test, insulin (0.75 IU/kg body weight; Humulin regular, Eli Lilly) was injected intraperitoneally into awake, fed control and transgenic mice. Bethesda, MD 20894, Copyright Glucose and insulin tolerance tests in KO and WT mice. Insulin tolerance test (ITT) Bring mice to lab around 5:30 PM Disinfect work area with 10% Clorox, then 70% EtOH Prepare work area with balance, clean bench paper, needles, blades, glucometer, and insulin syringes. For insulin tolerance test, mice are fasted for 5 hrs, and insulin (0.25 or 0.5 U/kg body weight) will be administered (ip or iv) into awake mice. Blood samples are taken at 10, 20, 30, 60, 90 and 120 min following injection for measurement of plasma glucose and insulin concentrations. Weigh mice, mark tails if necessary with different colors for rapid identification and take fasting glucose measurement via a tail clip. Please enable it to take advantage of the complete set of features! Keywords: Since most glucose is taken up into non-fat tissues, it can be confounding if you dose based on total body weight, and the two groups differ greatly with respect to the amount of fat mass. Accessibility 1-The mouse is given an intraperitoneal injection with a 27G needle of insulin. Insulin tolerance tests were done on fed (random) mice using an i.p. At the end of the glucose tolerance test, mice were returned to cages and allowed free access to food. Heliyon. In the present study, female C57BL/6J mice were maintained for four weeks under the condition of 8-hour phase advance and delay every 3–4 days to mimic shift work. AUC glucose (E) and AUC insulin (F) were calculated using the trapezoidal rule. 2020 Jan 20;11(1):5. doi: 10.1186/s13293-020-0281-3. Glucose; Insulin; Intraperitoneal injection; Metabolic test; Metabolism. A fat mouse and a lean mouse are likely to have very similar fat-free masses, and by giving more insulin to the fatter animal (because body weight is higher) you might make the fatter mouse seem more insulin sensitive, but really its just responding to a larger amount of insulin. Intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test (IPITT) were performed repeatedly at Zeitgeber time (ZT) 0, ZT6, ZT12, and ZT18. Diabetes Res Clin Pract. Sousa GM, Cazarin CBB, Maróstica Junior MR, Lamas CA, Quitete VHAC, Pastore GM, Bicas JL. If blood glucose drops below 10 mg/dL, or the animal appears catatonic, inject 300 uL of the glucose solution prepared above to rescue the mice and prevent hypoglycmic shock. To measure insulin sensitivity in the HFD fed mice, an insulin tolerance test was performed one week after the oral glucose tolerance test. This is a standard test to determine the insulin resistance status in humans and experimental animals. Reduced time points to calculate the composite index. After an overnight fast, an oral glucose load (1 mg/g) (A and B) or i.p. Weigh mice, mark tails if necessary with different colors for rapid identification and take fasting glucose measurement via a tail clip. Glucose concentration was determined in blood samples obtained from the tail vein at the indicated time points after the insulin … Typically starve the mice at 8AM and aim to start injections at 2PM. Abstract. Consecutive Low Doses of Streptozotocin Induce Polycystic Ovary Syndrome Features in Mice. 2011 Mar;26(3):419-20. doi: 10.1111/j.1440-1746.2010.06594.x. If you are using a higher or lower dose of insulin, add more or less to the 10 mL of PBS, so that injections are 10 uL/g of mass. Taking into Account Differences in Fat Mass, http://bridgeslab.sph.umich.edu/protocols/index.php?title=Insulin_Tolerance_Test&oldid=1619, Glucose Test Strips - AccuChek Comfort Curve or equivalent, echoMRI if the mice differ in body fat levels (see below). An insulin tolerance test (ITT) is a medical diagnostic procedure during … Use the R. This page was last modified on 8 June 2020, at 15:25. When measuring glucose just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse, which can be more stressful. The insulin tolerance test can be very dangerous because, if the pituitary gland or adrenal glands are not functioning properly, it can induce insulin shock and a coma from low blood glucose values. At desired intervals (normally 15, 30, 45, 60, 75, 90, 105 and 120 min), take blood glucose measurements from tail vein. doi: 10.1016/j.heliyon.2020.e03752. An ITT is usually ordered and interpreted by endocrinologists. Mice were fed their respective diets for 8 wk. doi: 10.1016/j.diabres.2011.08.025. 5 Glucose Tolerance Tests for Systematic Screening of Glucose Homeostasis in Mice At the end of the 6th and 7th week, an oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were respectively conducted in experiment A. Plasma glucose and insulin concentrations during the intraperitoneal glucose tolerance test (IPGTT; A and C) (2 g/kg) or oral glucose tolerance test (OGTT; B and D) (2 g/kg) following 6 h of fasting in C57BL/6J mice fed a standard chow ( ) or high-fat ( ) diet for 8 wk. Peng C, Xu X, Li Y, Li X, Yang X, Chen H, Zhu Y, Lu N, He C. Biol Sex Differ. Provide food and water as normal. The insulin tolerance test is commonly used in metabolic studies to assess whole body insulin sensitivity in rodents. The pyruvate bolus elicits a glycemic excursion that reflects hepatic gluconeogenesis. 25, 26 There are, nonetheless, important differences in the metabolic phenotypes of the 6J and 6N substrains of the C57BL/6 mice. In this chapter, we describe protocols to perform in the mouse glucose and insulin tolerance tests, two metabolic assays which evaluate the glucose tolerance and the insulin sensitivity, respectively. Animals were fasted overnight (12 h) and an oral glucose tolerance test (E) was performed after gavage with glucose (1 g/kg body weight). (D) Area under the curve for insulin tolerance tests. What is the recommended dose of insulin to perform insulin tolerance test in C57BL/6JUnib mice (fed on chow or high-fat diet)? It is performed on awake mice, fasted for 18 hr (overnight) or for 6 hours. An insulin tolerance test (ITT) is a medical diagnostic procedure during which insulin is injected into a patient's vein, after which blood glucose is measured at regular intervals. In vivo metabolic tests are highly valuable to determine whether atherosclerosis progression in mouse models is accompanied by carbohydrate metabolism alterations such as glucose intolerance and insulin resistance. 25,26 There are, nonetheless, important differences in the metabolic phenotypes of the 6J and 6N substrains of the C57BL/6 mice. Reproducibility of multiple repeated oral glucose tolerance tests. COVID-19 is an emerging, rapidly evolving situation. This is performed on C57Bl6, transgenic SENP1 KO and ZMIZ1 KO mouse models. FOIA At 0, 15, 30, 60, 90 and 120 min, blood was sampled (via the tail vein) for glucose determinations. Reagents and Materials: Protocol: 1. 2021 Jan 28;22(3):1299. doi: 10.3390/ijms22031299. Unable to load your collection due to an error, Unable to load your delegates due to an error. This will correspond to 1 U/kg injections. National Library of Medicine To test whether dietary AGEs play a role in the progression of insulin resistance in normal mice fed high-fat diets, normal C57/BL6 mice were randomly assigned to high-fat diets (35% g fat), either high (HAGE-HF group; 995.4 units/mg AGE) or low (by 2.4-fold LAGE-HF group; 329.6 … If needed re-snip the tail vein. 2020 Apr 30;6(4):e03752. 2020 Feb 14;21(4):1289. doi: 10.3390/ijms21041289. Add do not feed tag to cages, or ideally move cage to procedure room. Clipboard, Search History, and several other advanced features are temporarily unavailable. 2011 Dec;94(3):e78-82. Insulin tolerance test (ITT) The mice were administered with insulin (0.4 unit/kg, novolin, Novo Nordisk, Danmark) 4 h after fasting via intraperitoneal injection. For example, BALB/c mice display preserved glucose tolerance and insulin action under HFD, and this seems to be directly related to the lack of fat accumulation, despite lipid accumulation in the muscle. ( A , B ) Male WT BRJ and Kuma BRJ mice at 4-weeks-old (4w) ( A ) … Allow mice to rest in the lab O/N for ~ 15 hr. Try to make sure that the mice are in a quiet, undisturbed temperature controlled room with the lights on. The blood glucose levels were calculated using blood samples collected at 0, 40 and 90 min after insulin injection. The insulin is diluted from Humulin R-100 and is purchased through the veterinary staff. For this reason, it is important that the test be done directly by a doctor or other skilled healthcare professional who knows how to manage insulin shock. Int J Mol Sci. Should all patients with nonalcoholic fatty liver disease undergo oral glucose tolerance test? Remove mouse from cage and put into a clean In general you want the insulin to decrease blood glucose by about 60-70% in the most responsive of your too group so if your response is <20% of >70% change in blood glucose you will probably have to change your dose and retry. In this chapter, we describe protocols to perform in the mouse glucose and insulin tolerance tests, two metabolic assays which evaluate the glucose tolerance and the insulin sensitivity, … Sex-specific association between the gut microbiome and high-fat diet-induced metabolic disorders in mice. eCollection 2020 Apr. The mice are subjected to a seven-week HFD challenge using a Western diet. This may need to be adjusted depending on the insulin sensitivity of the mice, and this is based on a normal C57BL/6J mouse on chow. Prepare a 1 g/10mL solution of glucose in case some animals become hypoglyemic. Analyze data by both % change from fasting glucose and absolute values. Blood glucose levels were monitored from the tip of the tail vein with a glucometer (Accu-Chek; Roche, … 8600 Rockville Pike Would you like email updates of new search results? Remove food from mice for about 6h by putting them in a fresh cage. The Glucose Tolerance Test in Mice. Insulin Tolerance Test. It is a relatively simple test that involves measurement of blood glucose levels over time following a single intraperitoneal injection of insulin. Our preferred outcome is to report fasting glucose levels and report percent change as a figure, The preferred statistical model is a mixed linear model using the time points as ordinate values and testing for a main effect or an interaction of the treatment/genotype. 0.1 U/mL humulin in PBS (make as 10uL of 100 U/mL in 10 mL, sterile filtered). The effect of α-terpineol enantiomers on biomarkers of rats fed a high-fat diet. Insulin tolerance test The mice were injected intraperitoneally with a bolus of 0.5 U/kg insulin (NovoRapid 100 IU/ml) as previously described [ 11 ]. Gordon BA, Fraser SF, Bird SR, Benson AC. Fast mice for overnight or for 6 hours. The Insulin Tolerance Test (ITT) is designed to determine the whole body sensitivity of insulin receptors by measuring blood glucose levels changes before and after insulin administration. [Significance of insulin measurement in subjects with impaired glucose tolerance or type 2 diabetes]. Effects of Apocynin on Heart Muscle Oxidative Stress of Rats with Experimental Diabetes: Implications for Mitochondria. Careers. Fasting blood glucose (mmol/L) during the glucose tolerance test (GTT), the insulin tolerance test (ITT) and the respirometry at the four different ambient temperatures in male C57/6JBom mice after 6 weeks on control or high fat diet (HFD). Epub 2011 Sep 25. In general you want the insulin to decrease blood glucose by about 60-70% in the most responsive of your too group so if your response is <20% of >70% change in blood glucose you will probably have to change your dose and retry. Insulin Tolerance Test in Mouse. Int J Mol Sci. doi: 10.2337/dc10-0646. I have found 0.1 to 10 UI/kg in the literature. Compared to the LFD fed control group, the HFD fed mice showed an impaired reduction of blood glucose levels after insulin administration suggesting insulin resistance. A blood sample was drawn from the tail veins of conscious mice for measurements of serum glucose using a FreeStyle FLASH glucometer and strips (Abbott, Alameda, CA). Mice were 6 h fasted and an insulin tolerance test (C) was performed after an intraperitoneal injection of insulin (0.75 UI/kg body weight).

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