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Authors: Firuoz Danashgeri
Group: Diabetic Complications Consortium
Summary: This protocol describes the proper procedure for harvesting bladders from rats or mice for use by the DiaComp Uropathy group.Diabetic Complication:
Proper citation: Firuoz Danashgeri 2019. Removal and Preservation of the Urinary Bladder. protocols.io dx.doi.org/10.17504/protocols.io.8fahtie Copy
Authors: Firouz Daneshgari
Group: Diabetic Complications Consortium
Summary: 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
Proper citation: Firouz Daneshgari 2019. Conscious Cystometry Bladder Function Testing. protocols.io dx.doi.org/10.17504/protocols.io.3fegjje Copy
Authors: Kathi Burke, Peter Reifsnyder
Group: Diabetic Complications Consortium
Summary: Albuwell M is an indirect competitive ELISA designed to monitor kidney function in the mouse by measurement of urinary albumin. To complete the assay, sample and rabbit anti-murine albumin antibody are added to albumin coated wells. The antibody interacts and binds with the albumin immobilized to the stationary phase or with albumin in the fluid phase, hence the notion of competitive binding. A subsequent reaction with anti-rabbit -HRP conjugate labels the probe with enzyme. After washing, only the antibody-conjugate bound to the stationary phase remains in the well, and this is detected using a chromogenic reaction. Color intensity is inversely proportional to the logarithm of albumin in the fluid phase. The assay may be completed in less than 2.5 hoursDiabetic Complication:
Proper citation: Kathi Burke, Peter Reifsnyder 2019. Protocol for Albuwell M kit: Murine Microalbuminuria ELISA By Exocell Inc. protocols.io dx.doi.org/10.17504/protocols.io.34mgqu6 Copy
Authors: Nobuyo Maeda, Oliver Smithies, Nobuyuki Takahashi
Group: Diabetic Complications Consortium
Summary: SummaryIt is extremely important to measure quantitative parameters (e.g. plasma glucose, urinary albumin, etc.) in a way that minimizes the effects of any adventitious variables, such as seasonal changes, dietary differences, any differences in the degrees of backcrossing, or unplanned differences in genotypes. This means that, whenever possible, mice of the critical genotypes should be obtained from genotypically identical matings carried out at or close to the same time. Archival data are not appropriate. The breeding scheme to generate the experimental animals can vary in complexity, depending on the fertility of the two genders, whether the Testgene is to be studied in homozygotes or in heterozygotes, and whether the diabetes is induced genetically or by STZ. However, it is important to remember that, when looking for the effects of the Testgene genotype on diabetic complications, the aim is to compare the Testgene mutant/mutant (or Testgene mutant/wt) diabetic animals with their diabetic litter mates that are wildtype at the Testgene locus (Testgene wt/wt). ReferenceWang, C-H, Li, F, Hiller, S, Kim, H-S, Maeda, N, Smithies, O, and Takahashi, N. A modest decrease in endothelial NOS in mice comparable to that associated with human NOS3 variants exacerbates diabetic nephropathy. Proc. Natl. Acad. Sci. USA 108(5): 2070-2075 (2011) PMID: 21245338 PMCID: PMC3033253.Diabetic Complications:
Proper citation: Nobuyo Maeda, Oliver Smithies, Nobuyuki Takahashi 2019. Breeding Scheme and Selection of Animals for DiaComp Experiments. protocols.io dx.doi.org/10.17504/protocols.io.8fphtmn Copy
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