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| Name | Authors | DOI | Group |
Summary |
Associated Publications |
RRIDs used | ||||||
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WT-1 Staining Protocol for Podocytes Resource Report Resource Website |
Frank Brosius | DOI:10.17504/protocols.io.8e2htge | Diabetic Complications Consortium | This protocol describes a protocol used by some DiaComp members to detect podocyte nuclei in rodent glomeruli.Edited by: Brosius LaboratoryDiabetic Complications: | RRID:AB_2336820 | University of Arizona | https://diacomp.org/shared/document.aspx?id=63&docType=Protocol | 2 | 2019 | Frank Brosius 2019. WT-1 Staining Protocol for Podocytes. protocols.io https://dx.doi.org/10.17504/protocols.io.8e2htge | 2021-04-15 09:15:17 | |
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Neuropathy Phentoyping Protocols - Immuno-Electron Microscopy Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3jtgknn | Diabetic Complications Consortium | Summary:Phenotyping of Rodents for the Presence of Diabetic Neuropathy In man, the development of diabetic neuropathy is dependent on both the degree of glycemic control and the duration of diabetes. Diabetic neuropathy is a progressive disorder, with signs and symptoms that parallel the loss of nerve fibers over time. Consequently, assessments of neuropathy in mice are not performed at one time point, but are characterized at multiple time points during a 6 month period of diabetes. The degree of diabetes is evaluated in 2 ways: tail blood glucose measured following a 6 hour fast and glycated hemoglobin levels. The initial degree of neuropathy is screened using the methods discussed below. Detailed measures of neuropathy are employed when the initial screening instruments indicate a profound or unique phenotypic difference. This document contains protocols used by the DiaComp staff to examine and measure diabetic neuropathy at the whole animal, tissue and cellular levels.Diabetic Complication:Reference: Kumagai et al., Brain Research 1996, 706:313-317 | University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=54&docType=Protocol | 1 | 2019 | Eva Feldman 2019. Neuropathy Phentoyping Protocols - Immuno-Electron Microscopy. protocols.io https://dx.doi.org/10.17504/protocols.io.3jtgknn | 2021-04-15 09:15:24 | ||
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Intraperitoneal Glucose Tolerance Testing (IPGTT) Resource Report Resource Website |
Donald McClain | DOI:10.17504/protocols.io.3g7gjzn | Diabetic Complications Consortium | This is the standard protocol for most routine glucose tolerance testing. It is performed on awake mice, fasted for 6 hr.Diabetic Complications: | University of Utah | https://www.diacomp.org/shared/document.aspx?id=11&docType=Protocol | 1 | 2019 | Donald McClain 2019. Intraperitoneal Glucose Tolerance Testing (IPGTT). protocols.io https://dx.doi.org/10.17504/protocols.io.3g7gjzn | 2021-04-15 09:15:25 | ||
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Confocal imaging of live larval zebrafish for assessing peripheral neuropathy Resource Report Resource Website |
Sandra Rieger | DOI:10.17504/protocols.io.8i2huge | Diabetic Complications Consortium | This protocol describes a method to mount live 8 day-old zebrafish larvae in agarose for capturing fluorescent transgene-expressing peripheral somatosensory axons with confocal imaging. We have previously described a similar method in the Journal of Visualized Experiments (1).Diabetic Complication:References: 1. G. S. O'Brien, S. Rieger, S. M. Martin, A. M. Cavanaugh, C. Portera-Cailliau, A. Sagasti, Twophoton axotomy and time-lapse confocal imaging in live zebrafish embryos. J Vis Exp, (2009). 2. S. Rieger, A. Sagasti, Hydrogen peroxide promotes injury-induced peripheral sensory axon regeneration in the zebrafish skin. PLoS Biol 9, e1000621 (2011); published online EpubMay (10- PLBI-RA-9260R3 [pii] 10.1371/journal.pbio.1000621). 3. A. J. Pittman, M. Y. Law, C. B. Chien, Pathfinding in a large vertebrate axon tract: isotypic interactions guide retinotectal axons at multiple choice points. Development 135, 2865-2871 (2008); published online EpubSep | University of Miami | https://www.diacomp.org/shared/document.aspx?id=223&docType=Protocol | 2 | 2019 | Sandra Rieger 2019. Confocal imaging of live larval zebrafish for assessing peripheral neuropathy. protocols.io https://dx.doi.org/10.17504/protocols.io.8i2huge | 2021-04-15 09:15:36 | ||
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Glucose "Clamp" for measurement of whole body insulin-stimulated glucose disposal Resource Report Resource Website |
Donald McClain | DOI:10.17504/protocols.io.7s9hnh6 | Diabetic Complications Consortium | This is the standard protocol for measuring insulin action in terms of total body glucose disposal. It is the "gold standard" for quantifying insulin resistance/sensitivity.Diabetic Complications: | University of Utah | https://www.diacomp.org/shared/document.aspx?id=10&docType=Protocol | 2 | 2019 | Donald McClain 2019. Glucose "Clamp" for measurement of whole body insulin-stimulated glucose disposal. protocols.io https://dx.doi.org/10.17504/protocols.io.7s9hnh6 | 2021-04-15 09:15:53 | ||
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Induction of Type 1 Diabetes, Myocardial Infarction, Echocardiography, Measurement of Blood Glucose and Tissue Weights Resource Report Resource Website |
Michael Hill | DOI:10.17504/protocols.io.8fmhtk6 | Diabetic Complications Consortium | Summary:This protocol presented an induction of Type 1 Diabetes, and myocardial Infarction, performed an evaluation of in vivo residual LV function by echocardiography, and measurement of blood glucose and tissue weights of rats.Diabetic Complication: | University of Missouri - Columbia | https://www.diacomp.org/shared/document.aspx?id=267&docType=Protocol | 2 | 2019 | Michael Hill 2019. Induction of Type 1 Diabetes, Myocardial Infarction, Echocardiography, Measurement of Blood Glucose and Tissue Weights. protocols.io https://dx.doi.org/10.17504/protocols.io.8fmhtk6 | 2021-04-15 09:15:58 | ||
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Removal and Preservation of the Urinary Bladder Resource Report Resource Website |
Firuoz Danashgeri | DOI:10.17504/protocols.io.8fahtie | Diabetic Complications Consortium | This protocol describes the proper procedure for harvesting bladders from rats or mice for use by the DiaComp Uropathy group.Diabetic Complication: | Case Western Reserve University | https://www.diacomp.org/shared/document.aspx?id=30&docType=Protocol | 3 | 2019 | Firuoz Danashgeri 2019. Removal and Preservation of the Urinary Bladder. protocols.io https://dx.doi.org/10.17504/protocols.io.8fahtie | 2021-04-15 09:15:56 | ||
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Plantar Analgesia Test for Hind Paw and Tail Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.8a3hsgn | Diabetic Complications Consortium | This protocol measures and quantifies the animals sensitivity to pain in the hind paw. These measurements can be used to evaluate the pathology present in the nervous system.Diabetic Complication: | University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=37&docType=Protocol | 2 | 2019 | Eva Feldman 2019. Plantar Analgesia Test for Hind Paw and Tail. protocols.io https://dx.doi.org/10.17504/protocols.io.8a3hsgn | 2021-04-15 09:16:15 | ||
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Measurement of Left Ventricular Performance in Langendorff Perfused Mouse Hearts Resource Report Resource Website |
E. Dale Abel | DOI:10.17504/protocols.io.7tfhnjn | Diabetic Complications Consortium | Summary:This protocol describes the procedure used by the DiaComp for measuring left ventricular performance in isolated retrogradely perfused mouse hearts.Diabetic Complication: | University of Iowa | https://www.diacomp.org/shared/document.aspx?id=44&docType=Protocol | 2 | 2019 | E. Dale Abel 2019. Measurement of Left Ventricular Performance in Langendorff Perfused Mouse Hearts. protocols.io https://dx.doi.org/10.17504/protocols.io.7tfhnjn | 2021-04-15 09:16:13 | ||
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Neuropathy Phentoyping Protocols - Transmission Electron Microscopy Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3nzgmf6 | Diabetic Complications Consortium | Summary:Phenotyping of Rodents for the Presence of Diabetic Neuropathy In man, the development of diabetic neuropathy is dependent on both the degree of glycemic control and the duration of diabetes. Diabetic neuropathy is a progressive disorder, with signs and symptoms that parallel the loss of nerve fibers over time. Consequently, assessments of neuropathy in mice are not performed at one time point, but are characterized at multiple time points during a 6 month period of diabetes. The degree of diabetes is evaluated in 2 ways: tail blood glucose measured following a 6 hour fast and glycated hemoglobin levels. The initial degree of neuropathy is screened using the methods discussed below. Detailed measures of neuropathy are employed when the initial screening instruments indicate a profound or unique phenotypic difference. This document contains protocols used by the DiaComp staff to examine and measure diabetic neuropathy at the whole animal, tissue and cellular levels.Diabetic Complication: | University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=54&docType=Protocol | 1 | 2019 | Eva Feldman 2019. Neuropathy Phentoyping Protocols - Transmission Electron Microscopy. protocols.io https://dx.doi.org/10.17504/protocols.io.3nzgmf6 | 2021-04-15 09:16:26 | ||
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Neuropathy Phentoyping Protocols - Dosages of Tranquilizers and Anesthetics for Mice, Rats, Hamsters/Gerbils Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3jqgkmw | Diabetic Complications Consortium | Summary:Phenotyping of Rodents for the Presence of Diabetic Neuropathy In man, the development of diabetic neuropathy is dependent on both the degree of glycemic control and the duration of diabetes. Diabetic neuropathy is a progressive disorder, with signs and symptoms that parallel the loss of nerve fibers over time. Consequently, assessments of neuropathy in mice are not performed at one time point, but are characterized at multiple time points during a 6 month period of diabetes. The degree of diabetes is evaluated in 2 ways: tail blood glucose measured following a 6 hour fast and glycated hemoglobin levels. The initial degree of neuropathy is screened using the methods discussed below. Detailed measures of neuropathy are employed when the initial screening instruments indicate a profound or unique phenotypic difference. This document contains protocols used by the DiaComp staff to examine and measure diabetic neuropathy at the whole animal, tissue and cellular levels.Diabetic Complication: | University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=54&docType=Protocol | 1 | 2019 | Eva Feldman 2019. Neuropathy Phentoyping Protocols - Dosages of Tranquilizers and Anesthetics for Mice, Rats, Hamsters/Gerbils. protocols.io https://dx.doi.org/10.17504/protocols.io.3jqgkmw | 2021-04-15 09:16:25 | ||
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Serum, Blood and Body Weight Measurements in Pigs Resource Report Resource Website |
Tim Nichols, Dave Clemmons | DOI:10.17504/protocols.io.32agqae | Diabetic Complications Consortium | These assays are performed on all the pig models studied by the DiaComp to assess their serum chemistry and blood cell counts.Diabetic Complications: | University of North Carolina at Chapel Hill, University of North Carolina at Chapel Hill | https://www.diacomp.org/shared/document.aspx?id=51&docType=Protocol | 1 | 2019 | Tim Nichols, Dave Clemmons 2019. Serum, Blood and Body Weight Measurements in Pigs. protocols.io https://dx.doi.org/10.17504/protocols.io.32agqae | 2021-04-15 09:16:18 | ||
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Measurement of Left Ventricular Hemodynamic Parameters in Intact Mice Resource Report Resource Website |
E. Dale Abel | DOI:10.17504/protocols.io.7tdhni6 | Diabetic Complications Consortium | This protocol describes the procedure used by the DiaComp for measuring left ventricular hemodynamic parameters in intact mice.Diabetic Complication: | University of Iowa | https://www.diacomp.org/shared/document.aspx?id=43&docType=Protocol | 2 | 2019 | E. Dale Abel 2019. Measurement of Left Ventricular Hemodynamic Parameters in Intact Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.7tdhni6 | 2021-04-15 09:16:22 | ||
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Phenotypic Characterization of the Working Heart Resource Report Resource Website |
E. Dale Abel | DOI:10.17504/protocols.io.8a2hsge | Diabetic Complications Consortium | Summary:This protocol describes the procedure used by the DiaComp for cardiac performance, oxygen consumption and substrate metabolic rates in isolated working mouse hearts.Diabetic Complication:References:1. Belke, D.D., Larsen, T.S., Gibbs, E.M., Severson, D.L. 2001.Glucose metabolism in perfused mouse hearts overexpressing human GLUT-4 glucose transporter. Am J Physiol Endocrinol Metab.280:E420-7.2. Belke, D.D., Larsen, T.S., Gibbs, E.M., Severson, D.L. 2000.Altered metabolism causes cardiac dysfunction in perfused hearts from diabetic (db/db) mice. Am J Physiol Endocrinol Metab.279:E1104-13.3. Belke, D.D., Larsen, T.S., Lopaschuk, G.D., Severson, D.L. 1999.Glucose and fatty acid metabolism in the isolated working mouse heart. Am J Physiol.277:R1210-7.4. Larsen, T.S., Belke, D.D., Sas, R., Giles, W.R., Severson, D.L., Lopaschuk, G.D., Tyberg, J.V. 1999.The isolated working mouse heart: methodological considerations. Pflugers Arch.437:979-85.5. Belke D.D., Betuing S, Tuttle M.J., Graveleau C, Young M.E., Pham M., Zhang D., Cooksey R.C., McClain D.A., Litwin S.E., Taegtmeyer H., Severson D., Kahn C.R., Abel E.D. 2002. Insulin signaling coordinately regulates cardiac size, metabolism, and contractile protein isoform expression. J Clin Invest 109:629-639.6. Mazumder P.K., O’Neill B.T., Roberts M.W., Buchanan J., Yun U.J., Cooksey R.C., Boudina S., Abel E.D. 2004. Impaired Cardiac Efficiency and Increased Fatty Acid Oxidation in Insulin Resistant ob/ob mouse hearts. Diabetes. 53: 2366-2374.7. Suga H: Ventricular energetics. Physiol Rev 70:247-277, 1990 | University of Iowa | https://www.diacomp.org/shared/document.aspx?id=41&docType=Protocol | 2 | 2019 | E. Dale Abel 2019. Phenotypic Characterization of the Working Heart. protocols.io https://dx.doi.org/10.17504/protocols.io.8a2hsge | 2021-04-15 09:16:21 | ||
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Histological Evaluation of Renal Fibrosis in Mice Resource Report Resource Website |
Daisuke Katagiri, Takamune Takahashi | DOI:10.17504/protocols.io.3gygjxw | Diabetic Complications Consortium | Summary This protocol describes a protocol to evaluate histological fibrosis in mouse kidney. Diabetic Complication:References 1. Katagiri D, Hamasaki Y, Doi K, et al. Interstitial renal fibrosis due to multiple cisplatin treatments is ameliorated by semicarbazide-sensitive amine oxidase inhibition. Kidney Int 2015; 89: 374 - 385. 2. Hara S, Umeyama K, Yokoo T, et al. Diffuse glomerular nodular lesions in diabetic pigs carrying a dominant-negative mutant hepatocyte nuclear factor 1-alpha, an inheritant diabetic gene in humans. PLoS One 2014; 9: e92219. | RRID:AB_305584 | Vanderbilt University, Vanderbilt University | https://www.diacomp.org/shared/document.aspx?id=302&docType=Protocol | 1 | 2019 | Daisuke Katagiri, Takamune Takahashi 2019. Histological Evaluation of Renal Fibrosis in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.3gygjxw | 2021-04-15 09:16:37 | |
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Low-Dose Streptozotocin Induction Protocol (mouse) Resource Report Resource Website |
Frank Brosius | DOI:10.17504/protocols.io.8izhuf6 | Diabetic Complications Consortium | This protocol is used by DiaComp members to induce diabetes in a number of the animal models developed by the consortium. STZ is toxic to the insulinproducing beta cells of the pancreas and used to induce a diabetes similar to a type I diabetic (Reference). Some reports suggest cellular toxicity outside of the pancreas. STZ also exhibits broad spectrum antibacterial properties and alters the gut microbiota. Please ensure that appropriate controls are included in all studies and complementary models considered (e.g. the Ins2Akita mouse).Diabetic Complications: | University of Arizona | https://www.diacomp.org/shared/document.aspx?id=19&docType=Protocol | 2 | 2019 | Frank Brosius 2019. Low-Dose Streptozotocin Induction Protocol (mouse). protocols.io https://dx.doi.org/10.17504/protocols.io.8izhuf6 | 2021-04-15 09:16:48 | ||
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Determination of Podocyte Number and Density in Rodent Glomeruli Resource Report Resource Website |
Frank Brosius | DOI:10.17504/protocols.io.8eyhtfw | Diabetic Complications Consortium | This protocol describes a validated protocol used by some DiaComp members to determine podocyte numbers in rodent glomeruli.Edited by: Brosius LaboratoryDiabetic Complication:Reference: Sanden SK, Wiggins JE, Goyal M, Riggs LK, and Wiggins RC. Evaluation of a thick and thin section method for estimation of podocyte number, glomerular volume, and glomerular volume per podocyte in rat kidney with Wilms' tumor-1 protein used as a podocyte nuclear marker. J Am Soc Nephrol 14: 2484-2493, 2003. | University of Arizona | https://diacomp.org/shared/document.aspx?id=62&docType=Protocol | 2 | 2019 | Frank Brosius 2019. Determination of Podocyte Number and Density in Rodent Glomeruli. protocols.io https://dx.doi.org/10.17504/protocols.io.8eyhtfw | 2021-04-15 09:16:56 | ||
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Biochemical Measures of Neuropathy - Lowry Protein Assay Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3sggnbw | Diabetic Complications Consortium | Summary:Oxidative stress is highly correlated with the metabolic changes caused by hyperglycemia. Increased levels of glucose overload mitochondria and result in the production of reactive oxygen species (ROS). In addition, the flow of excess glucose through cellular pathways decreases the cell’s normal ability to detoxify ROS. As a result, the neurons and axons of the peripheral nervous system contain increased levels of ROS and decreased antioxidant capacity. The following assays are used to measure these changes in rodent models of diabetic neuropathy. Diabetic Complication: | University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=54&docType=Protocol | 1 | 2019 | Eva Feldman 2019. Biochemical Measures of Neuropathy - Lowry Protein Assay. protocols.io https://dx.doi.org/10.17504/protocols.io.3sggnbw | 2021-04-15 09:16:31 | ||
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Determination of Glomerular Filtration Rate in Conscious Mice using FITC-inulin Resource Report Resource Website |
Zhonghua Qi, Matthew D. Breyer | DOI:10.17504/protocols.io.3fngjme | Diabetic Complications Consortium | This protocol describes the procedures for estimating glomerular filtration rate in conscious mice based on Fluorescein Isothiocyanate-inulin (FITC-inulin) clearance following a single bolus intravenous injection. Diabetic Complication:Reference:1. Hem, A., Smith, A.J., and Solberg, P. 1998. Saphenous vein puncture for blood sampling of the mouse, rat, hamster, gerbil, guinea pig, ferret and mink. Lab Anim 32:364-368. 2. Sturgeon, C., Sam, A.D., 2nd, and Law, W.R. 1998. Rapid determination of glomerular filtration rate by single-bolus inulin: a comparison of estimation analyses. J Appl Physiol 84:2154-2162. 3. Qi, Z., Whitt, I., Mehta, A., Jin, J., Zhao, M., Harris, R.C., Fogo, A.B., and Breyer, M.D. 2003. Serial Determination of Glomerular Filtration Rate in Conscious Mice Using FITC-Inulin Clearance. Am J Physiol Renal Physiol. | Vanderbilt University, Eli Lilly Company | https://www.diacomp.org/shared/document.aspx?id=28&docType=Protocol | 1 | 2019 | Zhonghua Qi, Matthew D. Breyer 2019. Determination of Glomerular Filtration Rate in Conscious Mice using FITC-inulin. protocols.io https://dx.doi.org/10.17504/protocols.io.3fngjme | 2021-04-15 09:16:32 | ||
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Plasma Insulin (LINCO ELISA) Resource Report Resource Website |
Willa Hsueh, Alan Collins | DOI:10.17504/protocols.io.8a4hsgw | Diabetic Complications Consortium | This Rat / Mouse Insulin ELISA kit is used for the non radioactive quantification of insulin in mouse and rat sera. Plasma samples may also be used but application to samples of other biological fluids may need validation by the user.Diabetic Complications: | The Ohio State University College of Medicine, The Ohio State University College of Medicine | https://www.diacomp.org/shared/document.aspx?id=48&docType=Protocol | 2 | 2019 | Willa Hsueh, Alan Collins 2019. Plasma Insulin (LINCO ELISA). protocols.io https://dx.doi.org/10.17504/protocols.io.8a4hsgw | 2021-04-15 09:16:40 |
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