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Islet Studies

Distinct Effects of Saturated and Monounsaturated Fatty Acids on β-Cell Turnover and Function

  1. K. Maedler,
  2. G.A. Spinas,
  3. D. Dyntar,
  4. W. Moritz,
  5. N. Kaiser and
  6. Marc Y. Donath
  1. From the Division of Endocrinology and Diabetes (K.M., G.A.S., D.D., W.M., M.Y.D.), University Hospital, Zurich, Switzerland; and the Department of Endocrinology and Metabolism (N.K.), Hebrew University—Hadassah Medical Center, Jerusalem, Israel.
  1. Address correspondence and reprint requests to Marc Donath, MD, Division of Endocrinology and Diabetes, Department of Medicine, University Hospital, CH-8091 Zurich, Switzerland. E-mail: marc.donath{at}dim.usz.ch .
Diabetes 2001 Jan; 50(1): 69-76. https://doi.org/10.2337/diabetes.50.1.69
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  • FIG. 1.
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    FIG. 1.

    Characterization of the effect of palmitic acid on β-cell death by double staining with the TUNEL assay (alkaline phosphatase) and anti-insulin antibody (peroxidase) (A and B) and by double fluorescent staining with Annexin-V—FLUOS (green) (C and D) and propidium iodide (red) (E and F). Islets were plated on extracellular matrix-coated dishes and exposed for 4 days to media containing 11.1 mmol/l glucose alone (A, C, and E) and with 0.5 mmol/l palmitic acid (B, D, and F). The arrows indicate nuclei stained positive for the TUNEL reaction (light and fluorescence microscopy × 400).

  • FIG. 3.
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    FIG. 3.

    Fatty acids and glucose-induced β-cell DNA fragmentation and proliferative activity. Islets were cultured for 4 days in the absence or presence of different fatty acids (0.5 mmol/l) or a mixture of palmitoleic and palmitic acid (0.25 mmol/l [mixture] or 0.5 mmol/l [mixture 0.5] each) in 5.5, 11.1, and 33.3 mmol/l glucose. Results are means ± SE of the relative number of TUNEL+ (A) and Ki-67+ β-cells (B) per islet, normalized to control incubations at 5.5 mmol/l glucose alone (100%) (in absolute value: 0.82 TUNEL+ β-cells per islet and 4.06 Ki-67+ β-cells per islet at 5.5 mmol/l glucose alone). The mean number of islets scored for DNA fragmentation was 40, 68, and 49 and the mean number for proliferative activity by anti-Ki-67 staining was 78, 69, and 63 in media containing 5.5, 11.1, and 33.3 mmol/l glucose, respectively. Islets were isolated from 40 rats. *P < 0.01 between control and palmitic or palmitoleic acid at the same glucose concentration; §P < 0.05 between palmitic acid and fatty acid mixture at the same glucose concentration; **P < 0.01 relative to islets at 5.5 mmol/l glucose.

  • FIG. 2.
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    FIG. 2.

    Double immunostaining for the Ki-67 nuclear antigen (peroxidase) (A and B) and insulin (fluorescein) (C and D). Islets plated on extracellular matrix-coated dishes were exposed for 4 days to media containing 11.1 mmol/l glucose alone (A and C) or including 0.5 mmol/l palmitic acid (B and D). The arrows indicate nuclei stained positive for Ki-67 (light and fluorescence microscopy ×400).

  • FIG. 4.
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    FIG. 4.

    Effects of blockade of ceramide synthesis by fumonisin B1 and of exogenous ceramide on β-cell DNA fragmentation and proliferative activity. Islets were cultured for 4 days in 11.1 mmol/l glucose alone (control) or in the presence of 0.5 mmol/l palmitic acids with or without 15 μmol/l fumonisin B1 (Fumo) or in the presence of 15 μmol/l C2-ceramide. Results are means ± SE of the relative number of TUNEL+ (A) and Ki-67+ β-cells (B) per islet normalized to the solvent-treated control (100%) (in absolute value: 0.61 TUNEL+ β-cells per islet and 7.32 Ki-67+ β-cells per islet at 11.1 mmol/l glucose alone). The mean number of islets scored for DNA fragmentation and for proliferative activity by anti-Ki-67 staining was 85 and 71, respectively. Islets were isolated from 16 rats. *P < 0.01 relative to solvent-treated controls.

  • FIG. 5.
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    FIG. 5.

    Subcellular localization of ANT and cytochrome c in fatty acid-treated islets. Immunoblotting of ANT (molecular weight 30 kDa) and cytochrome c (molecular weight 15 kDa) was performed on mitochondrial (M) and cytosolic (C) fractions of islets cultured at 11.1 mmol/l glucose for 6 or 20 h with 0.5 mmol/l palmitic acid or 0.5 mmol/l palmitoleic acid. Control islets were analyzed after 20 h of incubation. Both antibodies were blotted on the same membrane after stripping. One representative of three experiments is shown.

  • FIG. 6.
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    FIG. 6.

    Confocal images showing cytochrome c diffusely localized to the cytoplasm but not significantly to the mitochondria in palmitic acid-treated islets. Islets were plated on extracellular matrix-coated dishes and were exposed for 4 days to media containing 11.1 mmol/l glucose alone (A) and with 0.5 mmol/l palmitic acid (B) and stained with anti-cytochrome c antibody.

  • FIG. 7.
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    FIG. 7.

    Effect of prolonged exposure of cultured islets to fatty acids on islet insulin content (A), on chronic insulin release into the culture medium (B), and on basal and glucose-stimulated insulin secretion (C). Islets were incubated in 11.1 mmol/l glucose in the absence or presence of 0.5 mmol/l palmitic or palmitoleic acid or a mixture of both (0.5 mmol/l each) for 4 days. Chronic insulin secretion represents the amount secreted into the culture medium during the experiment. Basal and stimulated insulin secretion denotes the amount secreted over 1 h incubation at 3.3 and 16.7 mmol/l glucose, respectively. Each bar represents the mean of 4 separate experiments ± SE. In each experiment, the data were collected from 4 plates per treatment. *P < 0.05 relative to solvent-treated controls; §P < 0.05 for the difference between palmitic acid and palmitoleic acid or fatty acid mixture.

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Distinct Effects of Saturated and Monounsaturated Fatty Acids on β-Cell Turnover and Function
K. Maedler, G.A. Spinas, D. Dyntar, W. Moritz, N. Kaiser, Marc Y. Donath
Diabetes Jan 2001, 50 (1) 69-76; DOI: 10.2337/diabetes.50.1.69

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Distinct Effects of Saturated and Monounsaturated Fatty Acids on β-Cell Turnover and Function
K. Maedler, G.A. Spinas, D. Dyntar, W. Moritz, N. Kaiser, Marc Y. Donath
Diabetes Jan 2001, 50 (1) 69-76; DOI: 10.2337/diabetes.50.1.69
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