Abstract
Swollen islet cells have been repeatedly described at onset of type 1 diabetes, but the underlying mechanism of this observation, termed hydropic degeneration, awaits characterization. In this study, laser capture microdissection was applied to extract the islets from an organ donor that died at onset of type 1 diabetes and from an organ donor without pancreatic disease. Morphologic analysis revealed extensive hydropic degeneration in 73 % of the islets from the donor with type 1 diabetes. Expression levels of genes involved in apoptosis, ER stress, beta cell function, and inflammation were analyzed in isolated and laser-captured islets by qPCR. The chemokine MCP-1 was expressed in islets from the donor with type 1 diabetes while undetectable in the control donor. No other signs of inflammation were detected. There were no signs of apoptosis on the gene expression level, which was also confirmed by negative immunostaining for cleaved caspase-8. There was an increased expression of the transcription factor ATF4, involved in transcription of ER stress genes, in the diabetic islets, but no further signs of ER stress were identified. In summary, on the transcription level, islets at onset of type 1 diabetes in which many beta cells display hydropic degeneration show no obvious signs of apoptosis, ER stress, or inflammation, supporting the notion that these cells are responding normally to high glucose and eventually succumbing to beta cell exhaustion. Also, this study validates the feasibility of performing qPCR analysis of RNA extracted from islets from subjects with recent onset of T1D and healthy controls by laser capture microdissection.
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Acknowledgments
This study was supported by grants from the Swedish Medical Research Council (65X-12219-15-6) and EU-FP7-Health 2010 PEVNET 261441. OK’s position is in part supported by the National Institutes of Health (2U01AI065192-06). Human pancreatic islets were obtained from The Nordic network for Clinical islet Transplantation, supported by the Swedish national strategic research initiative EXODIAB (Excellence Of Diabetes Research in Sweden) and the Juvenile Diabetes Research Foundation.
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Communicated by Antonio Secchi.
Johan Hopfgarten and Per-Anton Stenwall contributed equally and are listed in alphabetical order.
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Hopfgarten, J., Stenwall, PA., Wiberg, A. et al. Gene expression analysis of human islets in a subject at onset of type 1 diabetes. Acta Diabetol 51, 199–204 (2014). https://doi.org/10.1007/s00592-013-0479-5
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DOI: https://doi.org/10.1007/s00592-013-0479-5