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| Name | Authors | DOI | Group |
Summary |
Associated Publications |
RRIDs used | ||||||
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Podocyte Counting and Density Analysis Resource Report Resource Website |
Frank Brosius | DOI:10.17504/protocols.io.8a6hshe | Diabetic Complications Consortium | This protocol describes the procedures for quantifying the podocyte count and density in the glomerulus.Diabetic Complication: | University of Arizona | https://www.diacomp.org/shared/document.aspx?id=65&docType=Protocol | 2 | 2019 | Frank Brosius 2019. Podocyte Counting and Density Analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.8a6hshe | 2021-04-15 09:14:22 | ||
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Neuropathy Phentoyping Protocols - Apoptag (Peroxidase) Sections on Slides Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3jkgkkw | 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 - Apoptag (Peroxidase) Sections on Slides. protocols.io https://dx.doi.org/10.17504/protocols.io.3jkgkkw | 2021-04-15 09:14:21 | ||
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Biochemical Measures of Neuropathy - Hydrogen Peroxide Assay Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.7rghm3w | 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 | 2 | 2019 | Eva Feldman 2019. Biochemical Measures of Neuropathy - Hydrogen Peroxide Assay. protocols.io https://dx.doi.org/10.17504/protocols.io.7rghm3w | 2021-04-15 09:14:27 | ||
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Quantification of atherosclerosis at the aortic sinus Resource Report Resource Website |
Daniel Teupser, Jan Breslow | DOI:10.17504/protocols.io.3wsgpee | Diabetic Complications Consortium | This protocol describes the procedures for fixing, sectioning and staining the hearts of mice for signs of atherosclerosis. The following protocol covers each aspect of the procedure. Diabetic Complication: | Rockefeller University, Rockefeller University | https://www.diacomp.org/shared/document.aspx?id=16&docType=Protocol | 1 | 2019 | Daniel Teupser, Jan Breslow 2019. Quantification of atherosclerosis at the aortic sinus. protocols.io https://dx.doi.org/10.17504/protocols.io.3wsgpee | 2021-04-15 09:14:30 | ||
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NADH Oxidase Activity Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3jcgkiw | Diabetic Complications Consortium | Describes assay to quantitate NADH Oxidase activity from tissues. Diabetic Complications: | University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=36&docType=Protocol | 1 | 2019 | Eva Feldman 2019. NADH Oxidase Activity. protocols.io https://dx.doi.org/10.17504/protocols.io.3jcgkiw | 2021-04-15 09:14:45 | ||
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Conscious Cystometry Bladder Function Testing Resource Report Resource Website |
Firouz Daneshgari | DOI:10.17504/protocols.io.3fegjje | Diabetic Complications Consortium | Cystometry is a test of bladder function in which pressure and volume of fluid in the bladder is measured during filling, storage, and voiding. A cystometry study is performed to diagnose problems with urination, including incontinence and urinary retention.This test should be done on the mice two days after SPT Implantation.Diabetic Complication:Reference 1. Cannon TW, Damaser MS: Effects of anesthesia on cystometry and leak point pressure of the female rat. Life Sci 69:1193-1202, 2001 | Case Western Reserve University | https://www.diacomp.org/shared/document.aspx?id=52&docType=Protocol | 1 | 2019 | Firouz Daneshgari 2019. Conscious Cystometry Bladder Function Testing. protocols.io https://dx.doi.org/10.17504/protocols.io.3fegjje | 2021-04-15 09:14:46 | ||
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Neuropathy Phentoyping Protocols - In Situ Hybridization Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3m6gk9e | 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.This protocol was responsible by Dr. T. L. WoodDiabetic 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 - In Situ Hybridization. protocols.io https://dx.doi.org/10.17504/protocols.io.3m6gk9e | 2021-04-15 09:14:46 | ||
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Neuropathy Phentoyping Protocols - Streptozotocin Treatment For Rats Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3njgmcn | 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 - Streptozotocin Treatment For Rats. protocols.io https://dx.doi.org/10.17504/protocols.io.3njgmcn | 2021-04-15 09:14:34 | ||
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Echocardiography: Mouse Resource Report Resource Website |
E. Dale Abel | DOI:10.17504/protocols.io.7s7hnhn | Diabetic Complications Consortium | Summary:This protocol describes the procedure used by the DiaComp for transthoracic echocardiography in awake mice. Diabetic Complication: | University of Iowa | https://www.diacomp.org/shared/document.aspx?id=39&docType=Protocol | 2 | 2019 | E. Dale Abel 2019. Echocardiography: Mouse. protocols.io https://dx.doi.org/10.17504/protocols.io.7s7hnhn | 2021-04-15 09:14:49 | ||
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Creatinine Companion Protocol Assay (Exocell) Resource Report Resource Website |
Kathi Burke (Frank Brosius lab), Peter Reifsnyder (Ed Leiter Lab) | DOI:10.17504/protocols.io.3fmgjk6 | Diabetic Complications Consortium | This assay is designed for companion use with Exocell’s immunospecific ELISAs for urine albumin excretion, allowing expression of results as µg albumin per mg creatinine in the urine. Adaptation of creatinine measurement to microtiter wells enables convenient, simultaneous determination of albumin and creatinine concentrations in the same specimen, using analogous microtiter plate formats. Normalization of albumin excretion in relation to creatinine facilitates classification according to defined ranges of microalbuminuria, the established marker of the early diabetic renal dysfunction. The urine albumin-to-creatinine ration in a random urine specimen is an accepted alternative to cumbersome 24 urine collections in the detection and monitoring of microalbuminuria.The procedure is an adaptation of the alkaline picrate method and entails determination of the differential absorbance in a sample before and after the additions of acid to correct for color generation due to interfering substances.Diabetic Complication: | University of Arizona, Jackson Laboratory | https://www.diacomp.org/shared/document.aspx?id=23&docType=Protocol | 1 | 2019 | Kathi Burke (Frank Brosius lab), Peter Reifsnyder (Ed Leiter Lab) 2019. Creatinine Companion Protocol Assay (Exocell). protocols.io https://dx.doi.org/10.17504/protocols.io.3fmgjk6 | 2021-04-15 09:14:38 | ||
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Neuropathy Phentoyping Protocols - Intra-Epidermal Fiber Density Determination of Rodent Foot Pad Biopsies Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.7rehm3e | 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 | 2 | 2019 | Eva Feldman 2019. Neuropathy Phentoyping Protocols - Intra-Epidermal Fiber Density Determination of Rodent Foot Pad Biopsies. protocols.io https://dx.doi.org/10.17504/protocols.io.7rehm3e | 2021-04-15 09:15:00 | ||
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Dechorionation of zebrafish embryos with Pronase for metronidazole-mediated ß-cell ablation Resource Report Resource Website |
Sandra Rieger | DOI:10.17504/protocols.io.8exhtfn | Diabetic Complications Consortium | Zebrafish embryos develop in their eggshell (the chorion) until hatching (~48-72 hours post fertilization). Because the chorion is impermeable to water, it needs to be removed prior to metronidazole treatment and β-cell ablation in Tg(ins:NTR-mCherry) transgenic zebrafish embryos. This can be achieved either with forceps or by enzymatic removal using Pronase. The earliest time of removal should be after completion of gastrulation to avoid bursting of the yolk and potential infections.Diabetic Complication: | University of Miami | https://www.diacomp.org/shared/document.aspx?id=220&docType=Protocol | 2 | 2019 | Sandra Rieger 2019. Dechorionation of zebrafish embryos with Pronase for metronidazole-mediated ß-cell ablation. protocols.io https://dx.doi.org/10.17504/protocols.io.8exhtfn | 2021-04-15 09:15:07 | ||
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Ultracentrifugal separation of VLDL, LDL and HDL Resource Report Resource Website |
Daniel Teupser, Jan Breslow | DOI:10.17504/protocols.io.33bgqin | Diabetic Complications Consortium | This protocol is used to isolate the various lipid fractions from blood plasma using ultracentrifugation. The actual measured concentrations are performed separately once the isolations are complete.NOTE: This protocol is not applicable for ApoE knockout mice.Diabetic Complications: | Rockefeller University, Rockefeller University | https://www.diacomp.org/shared/document.aspx?id=18&docType=Protocol | 1 | 2019 | Daniel Teupser, Jan Breslow 2019. Ultracentrifugal separation of VLDL, LDL and HDL. protocols.io https://dx.doi.org/10.17504/protocols.io.33bgqin | 2021-04-15 09:14:57 | ||
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Ultracentrifugal separation of HDL alone and calculation of non-HDL Resource Report Resource Website |
Daniel Teupser, Jan Breslow | DOI:10.17504/protocols.io.32mgqc6 | Diabetic Complications Consortium | This protocol is used to isolate the various lipid fractions from blood plasma using ultracentrifugation. The actual measured concentrations are performed separately once the isolations are complete.NOTE: This protocol IS applicable for ApoE knockout mice.Diabetic Complication: | Rockefeller University, Rockefeller University | https://www.diacomp.org/shared/document.aspx?id=17&docType=Protocol | 1 | 2019 | Daniel Teupser, Jan Breslow 2019. Ultracentrifugal separation of HDL alone and calculation of non-HDL. protocols.io https://dx.doi.org/10.17504/protocols.io.32mgqc6 | 2021-04-15 09:15:30 | ||
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Neuropathy Phentoyping Protocols - Immunofluorescence Method for 8DG Localizaiton Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3jwgkpe | 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:Yarborough et al, Cancer Res. 56:683-688, 1996 | University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=54&docType=Protocol | 1 | 2019 | Eva Feldman 2019. Neuropathy Phentoyping Protocols - Immunofluorescence Method for 8DG Localizaiton. protocols.io https://dx.doi.org/10.17504/protocols.io.3jwgkpe | 2021-04-15 09:15:01 | ||
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Pancreatic Insulin Content by Acid-Ethanol Extraction Resource Report Resource Website |
Ed Leiter | DOI:10.17504/protocols.io.3jjgkkn | Diabetic Complications Consortium | Used to calculate the insulin content in the pancreas.Diabetic Complications: | Jackson Laboratories | https://www.diacomp.org/shared/document.aspx?id=73&docType=Protocol | 1 | 2019 | Ed Leiter 2019. Pancreatic Insulin Content by Acid-Ethanol Extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.3jjgkkn | 2021-04-15 09:15:10 | ||
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Biochemical Measures of Neuropathy - Genotyping Resource Report Resource Website |
Eva Feldman | DOI:10.17504/protocols.io.3qdgms6 | 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 - Genotyping. protocols.io https://dx.doi.org/10.17504/protocols.io.3qdgms6 | 2021-04-15 09:15:13 | ||
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Atherosclerotic Morphometry in Pigs Resource Report Resource Website |
Tim Nichols, Dave Clemmons | DOI:10.17504/protocols.io.7s6hnhe | Diabetic Complications Consortium | These assays are performed on all the pig models studied by the DiaComp to assess the extent of atherosclerotic lesions in diabetic cardiovascular disease models. Each measurement/assay is listed below with the details of how the data is collected. Details of the reagents and materials are in the assay.Diabetic Complication: | University of North Carolina at Chapel Hill, University of North Carolina at Chapel Hill | https://www.diacomp.org/shared/document.aspx?id=49&docType=Protocol | 2 | 2019 | Tim Nichols, Dave Clemmons 2019. Atherosclerotic Morphometry in Pigs. protocols.io https://dx.doi.org/10.17504/protocols.io.7s6hnhe | 2021-04-15 09:15:13 | ||
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Breeding NONcNZO10/LtJ (NON/NZ) and NON/ShiLtJ (NON/Shi) mice Resource Report Resource Website |
Ian Simpson | DOI:10.17504/protocols.io.556g89e | Diabetic Complications Consortium | ♦ NONcNZO10/LtJ (NON/NZ) mice are the mice used for a Type 2 diabetes model. The mice are given 5K20 diet at weaning and the mice are usually exhibiting hyperglycemia by 10-11 weeks of age (see Figure 1) 90- 95 % of the mice will become diabetic at 20 weeks. ♦ NON/ShiLtJ (NON/Shi) mice are the mice used for a control to the diabetic mice. The mice are given 5K20 diet at weaning and should not become diabetic. But 5-10% of the mice will become diabetic at 20 weeks. ♦ Breeding, housing, and experimentation are performed in a pathogen-free environmentDiabetic Complications: | Pennsylvania State University | https://www.diacomp.org/shared/document.aspx?id=250&docType=Protocol | 2 | 2019 | Ian Simpson 2019. Breeding NONcNZO10/LtJ (NON/NZ) and NON/ShiLtJ (NON/Shi) mice. protocols.io https://dx.doi.org/10.17504/protocols.io.556g89e | 2021-04-15 09:15:40 | ||
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Neuropathy Phentoyping Protocols - Insulin Implantation Protocol Resource Report Resource Website |
Eva Feldman, Omorodola Abatan | DOI:10.17504/protocols.io.7rdhm26 | 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, University of Michigan - Ann Arbor | https://www.diacomp.org/shared/document.aspx?id=54&docType=Protocol | 2 | 2019 | Eva Feldman, Omorodola Abatan 2019. Neuropathy Phentoyping Protocols - Insulin Implantation Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7rdhm26 | 2021-04-15 09:15:16 |
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