Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells
Abstract Background Rodent and human β-cells are differentially susceptible to the “lipotoxic” effects of long-chain saturated fatty acids (LC-SFA) but the factors accounting for this are unclear. Here, we have studied the intracellular disposition of the LC-SFA palmitate in human vs rodent β–cells...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Publishing Group
2022-04-01
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| Series: | Nutrition & Diabetes |
| Online Access: | https://doi.org/10.1038/s41387-022-00199-y |
| _version_ | 1828347192358731776 |
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| author | Patricia Thomas Catherine Arden Jenna Corcoran Christian Hacker Hannah J. Welters Noel G. Morgan |
| author_facet | Patricia Thomas Catherine Arden Jenna Corcoran Christian Hacker Hannah J. Welters Noel G. Morgan |
| author_sort | Patricia Thomas |
| collection | DOAJ |
| description | Abstract Background Rodent and human β-cells are differentially susceptible to the “lipotoxic” effects of long-chain saturated fatty acids (LC-SFA) but the factors accounting for this are unclear. Here, we have studied the intracellular disposition of the LC-SFA palmitate in human vs rodent β–cells and present data that reveal new insights into the factors regulating β-cell lipotoxicity. Methods The subcellular distribution of the LC-SFA palmitate was studied in rodent (INS-1E and INS-1 823/13 cells) and human (EndoC-βH1) β-cells using confocal fluorescence and electron microscopy (EM). Protein expression was assessed by Western blotting and cell viability, by vital dye staining. Results Exposure of INS-1 cells to palmitate for 24 h led to loss of viability, whereas EndoC-βH1 cells remained viable even after 72 h of treatment with a high concentration (1 mM) of palmitate. Use of the fluorescent palmitate analogue BODIPY FL C16 revealed an early localisation of the LC-SFA to the Golgi apparatus in INS-1 cells and this correlated with distention of intracellular membranes, visualised under the EM. Despite this, the PERK-dependent ER stress pathway was not activated under these conditions. By contrast, BODIPY FL C16 did not accumulate in the Golgi apparatus in EndoC-βH1 cells but, rather, co-localised with the lipid droplet-associated protein, PLIN2, suggesting preferential routing into lipid droplets. When INS-1 cells were treated with a combination of palmitate plus oleate, the toxic effects of palmitate were attenuated and BODIPY FL C16 localised primarily with PLIN2 but not with a Golgi marker. Conclusion In rodent β-cells, palmitate accumulates in the Golgi apparatus at early time points whereas, in EndoC- βH1 cells, it is routed preferentially into lipid droplets. This may account for the differential sensitivity of rodent vs human β-cells to “lipotoxicity” since manoeuvres leading to the incorporation of palmitate into lipid droplets is associated with the maintenance of cell viability in both cell types. |
| first_indexed | 2024-04-14T00:36:28Z |
| format | Article |
| id | doaj.art-07ad9c44b3774e06ab8db3c7f113528e |
| institution | Directory Open Access Journal |
| issn | 2044-4052 |
| language | English |
| last_indexed | 2024-04-14T00:36:28Z |
| publishDate | 2022-04-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Nutrition & Diabetes |
| spelling | doaj.art-07ad9c44b3774e06ab8db3c7f113528e2022-12-22T02:22:21ZengNature Publishing GroupNutrition & Diabetes2044-40522022-04-011211810.1038/s41387-022-00199-yDifferential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cellsPatricia Thomas0Catherine Arden1Jenna Corcoran2Christian Hacker3Hannah J. Welters4Noel G. Morgan5Institute of Metabolism and Systems Research, Birmingham Medical School, University of BirminghamBiosciences Institute, The Medical School, Newcastle UniversityDepartment of Biosciences, University of Exeter, Geoffrey Pope BuildingBioimaging Centre, College of Life and Environmental Sciences, University of ExeterInstitute of Biomedical and Clinical Research, College of Medicine and Health, University of ExeterInstitute of Biomedical and Clinical Research, College of Medicine and Health, University of ExeterAbstract Background Rodent and human β-cells are differentially susceptible to the “lipotoxic” effects of long-chain saturated fatty acids (LC-SFA) but the factors accounting for this are unclear. Here, we have studied the intracellular disposition of the LC-SFA palmitate in human vs rodent β–cells and present data that reveal new insights into the factors regulating β-cell lipotoxicity. Methods The subcellular distribution of the LC-SFA palmitate was studied in rodent (INS-1E and INS-1 823/13 cells) and human (EndoC-βH1) β-cells using confocal fluorescence and electron microscopy (EM). Protein expression was assessed by Western blotting and cell viability, by vital dye staining. Results Exposure of INS-1 cells to palmitate for 24 h led to loss of viability, whereas EndoC-βH1 cells remained viable even after 72 h of treatment with a high concentration (1 mM) of palmitate. Use of the fluorescent palmitate analogue BODIPY FL C16 revealed an early localisation of the LC-SFA to the Golgi apparatus in INS-1 cells and this correlated with distention of intracellular membranes, visualised under the EM. Despite this, the PERK-dependent ER stress pathway was not activated under these conditions. By contrast, BODIPY FL C16 did not accumulate in the Golgi apparatus in EndoC-βH1 cells but, rather, co-localised with the lipid droplet-associated protein, PLIN2, suggesting preferential routing into lipid droplets. When INS-1 cells were treated with a combination of palmitate plus oleate, the toxic effects of palmitate were attenuated and BODIPY FL C16 localised primarily with PLIN2 but not with a Golgi marker. Conclusion In rodent β-cells, palmitate accumulates in the Golgi apparatus at early time points whereas, in EndoC- βH1 cells, it is routed preferentially into lipid droplets. This may account for the differential sensitivity of rodent vs human β-cells to “lipotoxicity” since manoeuvres leading to the incorporation of palmitate into lipid droplets is associated with the maintenance of cell viability in both cell types.https://doi.org/10.1038/s41387-022-00199-y |
| spellingShingle | Patricia Thomas Catherine Arden Jenna Corcoran Christian Hacker Hannah J. Welters Noel G. Morgan Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells Nutrition & Diabetes |
| title | Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells |
| title_full | Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells |
| title_fullStr | Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells |
| title_full_unstemmed | Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells |
| title_short | Differential routing and disposition of the long-chain saturated fatty acid palmitate in rodent vs human beta-cells |
| title_sort | differential routing and disposition of the long chain saturated fatty acid palmitate in rodent vs human beta cells |
| url | https://doi.org/10.1038/s41387-022-00199-y |
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