Transcutaneous measurement of glomerular filtration rate using FITC-sinistrin in rats

Schock-Kusch, Daniel; Sadick, Maliha; Henninger, Nadja; Kraenzlin, Bettina; Claus, Guenter; Kloetzer, Hans-Martin; Weiß, Christel; Pill, Johannes; Gretz, Norbert

doi:10.1093/ndt/gfp225

Cited by 122 publications

(133 citation statements)

References 21 publications

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“…One of them is the independence from blood and urine sampling which leads to great savings in terms of consumables, time and, of course, animal stress. Table 2, the exogenous renal marker FITC-sinistrin and the measurement of its elimination kinetics through the skin allow the impact of renal pathologies on kidney function to be studied 9,10,[14][15][16][17][18] . The serial collection of blood samples required to assess plasma clearance or biomarkers of renal disease are stressful for the animal, cumbersome and time-consuming for the researchers.…”

Section: Discussionmentioning

confidence: 99%

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Transcutaneous Assessment of Renal Function in Conscious Rodents

Pérez

Weinfurter

Gretz

2016

JoVE

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Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and time-consuming. In addition, serial blood or urine samples are required, with the associated stress for the experimental animals. A recent technique significantly reduces the investment in time and resources, minimizing the invasiveness and the animal stress, but being equally valid as the traditional approaches. The method measures transcutaneously renal function. Using an optical device and the exogenous renal marker fluorescein isothiocyanate (FITC)-sinistrin, this technique is capable of measuring the elimination kinetics of the marker through the skin. With neither blood nor urine samples nor the associated laboratory assays needed, the results of the transcutaneous measurement are almost instantaneously available. The method has been already validated in different species and successfully applied in several models of renal pathology. Moreover, due to its minimally invasive characteristics, it is suitable for sequential measurements within the same animal. Here is provided a detailed protocol to carry out the transcutaneous assessment of renal function in rodents.

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Section: Discussionmentioning

confidence: 99%

“…Note: The software will automatically display the FITC-sinistrin half-life (t 1/2 ) along with an R 2 value, determined by a 1-compartment model. t 1/2 can be utilized to calculate GFR by using a conversion factor 9,10 .…”

Section: Read Out Of Datamentioning

confidence: 99%

Transcutaneous Assessment of Renal Function in Conscious Rodents

Pérez

Weinfurter

Gretz

2016

JoVE

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“…Note: if using FITC-sinistrin [16][17][18] , which does not require dialysis, all steps required for dialysis can be skipped.…”

Section: Preparation Of Fitc-inulin Injection Solutionmentioning

confidence: 99%

A High-throughput Method for Measurement of Glomerular Filtration Rate in Conscious Mice

Rieg

2013

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The measurement of glomerular filtration rate (GFR) is the gold standard in kidney function assessment. Currently, investigators determine GFR by measuring the level of the endogenous biomarker creatinine or exogenously applied radioactive labeled inulin ( 3 H or 14 C). Creatinine has the substantial drawback that proximal tubular secretion accounts for ~50% of total renal creatinine excretion and therefore creatinine is not a reliable GFR marker. Depending on the experiment performed, inulin clearance can be determined by an intravenous single bolus injection or continuous infusion (intravenous or osmotic minipump). Both approaches require the collection of plasma or plasma and urine, respectively. Other drawbacks of radioactive labeled inulin include usage of isotopes, time consuming surgical preparation of the animals, and the requirement of a terminal experiment. Here we describe a method which uses a single bolus injection of fluorescein isothiocyanate-(FITC) labeled inulin and the measurement of its fluorescence in 1-2 μl of diluted plasma. By applying a two-compartment model, with 8 blood collections per mouse, it is possible to measure GFR in up to 24 mice per day using a special work-flow protocol. This method only requires brief isoflurane anesthesia with all the blood samples being collected in a non-restrained and awake mouse. Another advantage is that it is possible to follow mice over a period of several months and treatments (i.e. doing paired experiments with dietary changes or drug applications). We hope that this technique of measuring GFR is useful to other investigators studying mouse kidney function and will replace less accurate methods of estimating kidney function, such as plasma creatinine and blood urea nitrogen.

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“…previously described (12,13). In short, pups were briefly sedated by inhalation of 5% isoflurane, followed by a single bolus injection of FITC-sinistrin (7 mg/100 g B.W.)…”

Section: Tunel Stainingmentioning

confidence: 99%

Low birth weight is associated with impaired murine kidney development and function

et al. 2017

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BACKGROUND: Low birth weight (LBW) neonates have impaired kidney development that leaves them susceptible to kidney disease and hypertension during adulthood. The study here identifies events that blunt nephrogenesis and kidney development in the murine LBW neonate. METHODS: We examined survival, kidney development, GFR, gene expression, and cyto-/chemokines in the LBW offspring of malnourished (caloric and protein-restricted) pregnant mice. RESULTS: Malnourished pregnant mothers gave birth to LBW neonates that had 40% reduced body weight and 54% decreased survival. Renal blood perfusion was reduced by 37%, whereas kidney volume and GFR were diminished in the LBW neonate. During gestation, the LBW neonatal kidney had 2.2-fold increased apoptosis, 76% decreased SIX2+ progenitor cells, downregulation of mesenchymal-to-epithelial signaling factors Wnt9b and Fgf8, 64% less renal vesicle formation, and 32% fewer nephrons than controls. At birth, increased plasma levels of IL-1β, IL-6, IL-12(p70), and granulocyte-macrophage colony-stimulating factor in the LBW neonate reduced SIX2+ progenitor cells. CONCLUSION: Increased pro-inflammatory cytokines in the LBW neonate decrease SIX2+ stem cells in the developing kidney. Reduced renal stem cells (along with the decreased mesenchymal-to-epithelial signaling) blunt renal vesicle generation, nephron formation, and kidney development. Subsequently, the mouse LBW neonate has reduced glomeruli volume, renal perfusion, and GFR.

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Transcutaneous measurement of glomerular filtration rate using FITC-sinistrin in rats

Cited by 122 publications

References 21 publications

Transcutaneous Assessment of Renal Function in Conscious Rodents

Transcutaneous Assessment of Renal Function in Conscious Rodents

A High-throughput Method for Measurement of Glomerular Filtration Rate in Conscious Mice

Low birth weight is associated with impaired murine kidney development and function

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