Plasma creatinine determination in mice and rats: An enzymatic method compares favorably with a high-performance liquid chromatography assay

Keppler, Andrea; Gretz, Norbert; Schmidt, Ralph Erich; Hm, Kloetzer; Hj, Groene; Lelongt, Brigitte; Meyer, Mary C.; Sadick, Maliha; Pill, Johannes

doi:10.1038/sj.ki.5001988

Cited by 114 publications

(100 citation statements)

References 10 publications

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“…Another limitation is that serum creatinine is below detection limits of several commercially available assays compared to HPLC determination. However, by using enzymatic reactions investigators were able to determine plasma creatinine in mice 10,22 . GFR in a similar range were found in conscious mice using a comparable single-bolus injection method as well as urinary FITC-inulin clearance 14 .…”

Section: Discussionmentioning

confidence: 99%

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

Rieg

2013

JoVE

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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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Section: Discussionmentioning

confidence: 99%

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

Rieg

2013

JoVE

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“…Plasma and urinary creatinine were determined using an enzymatic method that has been validated in rodents. 40 Creatinine clearance (ml/min) was derived from the following formula: urinary creatinine 3 urine volume 3 1440 min -1 3 plasma creatinine -1…”

Section: Creatinine Clearance Determinationmentioning

confidence: 99%

Cellular Senescence Limits Regenerative Capacity and Allograft Survival

Braun

Schmidt

Raiss

et al. 2012

Journal of the American Society of Nephrology

146

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Long-term graft survival after kidney transplantation remains unsatisfactory and unpredictable. Interstitial fibrosis and tubular atrophy are major contributors to late graft loss; features of tubular cell senescence, such as increased p16INK4a expression, associate with these tubulointerstitial changes, but it is unknown whether the relationship is causal. Here, loss of the INK4a locus in mice, which allows escape from p16 INK4a -dependent senescence, significantly reduced interstitial fibrosis and tubular atrophy and associated with improved renal function, conservation of nephron mass, and transplant survival. Compared with wild-type controls, kidneys from INK4a 2/2 mice developed significantly less interstitial fibrosis and tubular atrophy after ischemia-reperfusion injury. Consistently, mice that received kidney transplants from INK4a/ARF 2/2 donors had significantly better survival 21 days after life-supporting kidney transplantation and developed less tubulointerstitial changes. This correlated with higher proliferative rates of tubular cells and significantly fewer senescent cells. Taken together, these data suggest a pathogenic role of renal cellular senescence in the development of interstitial fibrosis and tubular atrophy and kidney graft deterioration by preventing the recovery from injury. Inhibiting premature senescence could have therapeutic benefit in kidney transplantation but has to be balanced against the risks of suspending antitumor defenses.

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“…Albumin and creatinine measurements were performed on a Cobas Fara centrifugal analyzer (Roche Diagnostics Ltd, Welwyn Garden City, UK) using a commercial immunoturbidimetric assay (Microalbumin Kit, Olympus Diagnostics Ltd, Watford, UK) and a creatininase-based enzymatic method 26 (Alpha Laboratories Ltd, Eastleigh, UK), respectively.…”

Section: Albumin and Creatinine Measurementsmentioning

confidence: 99%

Hyperglycemia and Renin-Dependent Hypertension Synergize to Model Diabetic Nephropathy

Conway

Rennie

Bailey

et al. 2012

Journal of the American Society of Nephrology

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Rodent models exhibit only the earliest features of human diabetic nephropathy, which limits our ability to investigate new therapies. Hypertension is a prerequisite for advanced diabetic nephropathy in humans, so its rarity in typical rodent models may partly explain their resistance to nephropathy. Here, we used the Cyp1a1mRen2 rat, in which the murine renin-2 gene is incorporated under the Cytochrome P4501a1 promoter. In this transgenic strain, administration of low-dose dietary indole-3-carbinol induces moderate hypertension. In the absence of hypertension, streptozotocininduced diabetes resulted in a 14-fold increase in albuminuria but only mild changes in histology and gene expression despite 28 weeks of marked hyperglycemia. In the presence of induced hypertension, hyperglycemia resulted in a 500-fold increase in albuminuria, marked glomerulosclerosis and tubulointerstitial fibrosis, and induction of many of the same pathways that are upregulated in the tubulointerstitium in human diabetic nephropathy. In conclusion, although induction of diabetes alone in rodents has limited utility to model human diabetic nephropathy, renin-dependent hypertension and hyperglycemia synergize to recapitulate many of the clinical, histological, and gene expression changes observed in humans.

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Plasma creatinine determination in mice and rats: An enzymatic method compares favorably with a high-performance liquid chromatography assay

Cited by 114 publications

References 10 publications

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

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

Cellular Senescence Limits Regenerative Capacity and Allograft Survival

Hyperglycemia and Renin-Dependent Hypertension Synergize to Model Diabetic Nephropathy

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