Volume 55, Issue 4, Pages (April 1999)

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Volume 55, Issue 4, Pages 1293-1302 (April 1999) Mesangial cells from diabetic NOD mice constitutively express increased density of atrial natriuretic peptide C receptors Nicole Ardaillou, Sandrine Placier, Liliane Striker, Gary Striker, Raymond Ardaillou Kidney International Volume 55, Issue 4, Pages 1293-1302 (April 1999) DOI: 10.1046/j.1523-1755.1999.00393.x Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 1 The cGMP response of nondiabetic nonobese diabetic (ND-NOD) and diabetic nonobese diabetic (D-NOD) mice mesangial cells to increasing concentrations of atrial natriuretic peptide (ANP;A) or C-type natriuretic peptide (CNP;B). cGMP was measured after a five-minute incubation period at 37°C. Cells were cultured during at least two weeks before the experiment in medium containing 6mm (•) or 20mm (▾) glucose. Data are means ± se (five experiments with triplicates in each) are given. Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 2 The time course of specific binding of [125I]-ANP to ND-NOD (▾) and D-NOD (•) mesangial cells at 4°C. Specific binding was measured during incubation with 50 pm [125I]-ANP. Data are means ± se (two experiments with triplicates in each). Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 3 Saturation binding study of specific [125I]-ANP binding to ND-NOD (▾) and D-NOD (•) mesangial cells. (A) A representative curve performed with triplicates for each point is shown. (B) Scatchard transformation of the data is shown with the calculated values of KD = 92.5 pm fmol/mg and Bmax = 540 fmol/mg for D-NOD, and KD = 89.2 pm fmol/mg and Bmax = 118 fmol/mg for ND-NOD. Five additional studies were performed. Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 4 Saturation binding study of specific [125I]-ANP binding to ND-NOD and D-NOD mesangial cells. Cultures were maintained during at least two weeks in medium containing 6mm (•) or 20mm (▾) glucose. Means ± se of two experiments with triplicates in each are given. Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 5 Competitive inhibition of [125I]-ANP binding to D-NOD (A) and ND-NOD (B) mesangial cells in the presence of increasing concentrations of unlabeled ANP (•) or 4-23C ANP (▾). Means ± se values (two experiments with triplicates in each) are given. The results are expressed as percentage of control. Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 6 Time course of binding of [125I]-ANP to ND-NOD (▾) and D-NOD (•) mesangial cells at 37°C. (A) Specific binding. (B) Cell-associated radioactivity was determined after the acidic wash of the cells. Means ± se (two experiments with triplicate in each) are given. Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 7 The NPR-C mRNA levels in ND-NOD and D-NOD mesangial cells determined by RNase protection assay. A typical autoradiogram of polyacrylamide gel electrophoresis with samples of 5 to 20 μg of RNA is shown. The ratio of NPR-C over actin mRNA was greater in D-NOD than in ND-NOD cells with a multiplication factor of 3.3 ± 0.7 (mean ± se of three experiments). Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 8 The peptide C receptor (PCR) products obtained after amplification of ND-NOD and D-NOD cell cDNA. Amplification products specific for NPR-A and NPR-C were obtained. A DNA molecular weight ladder is shown. The results of three experiments are shown. Kidney International 1999 55, 1293-1302DOI: (10.1046/j.1523-1755.1999.00393.x) Copyright © 1999 International Society of Nephrology Terms and Conditions