Biochemical and Biophysical Research Communications
Glucagon production of the rat insulinoma cell line INS-1—A quantitative comparison with primary rat pancreatic islets☆
Section snippets
Materials and methods
Reagents. Radioimmunoassays for rat pancreatic glucagon (crossreactivity to oxyntomodulin <0.1%) and for immunoreactive insulin (IRI) were purchased from Linco Research (St. Louis, USA). For immunofluorescence, the antiserum (derived from guinea-pig) against rat c-peptide was from Linco Research (St. Louis, USA), the specific monoclonal antibody (derived from mouse) against rat glucagon was from Sigma–Aldrich (Taufkirchen, Germany), the secondary guinea-pig immunoglobulin antibody conjugated to
INS-1 cells release both insulin and glucagon in relevant amounts
The insulin and glucagon release of INS-1 cells (n = 5 experiments, each containing four independent samples in parallel) and rat pancreatic islets (n = 3 experiments, each containing four independent samples in parallel) was monitored over 150 min with glucose concentrations increasing stepwise from 1.4 to 22.4 mM. The secretion values for immunoreactive insulin and glucagon were normalized by the cellular protein content.
Before exploring a putative α-cellular behavior of the INS-1 cells, we first
Conclusions
We found a substantial glucagon secretion from the polyclonal rat insulinoma cell line INS-1. While the intracellular presence of glucagon could be qualitatively confirmed by immunofluorescence experiments, the intracellular glucagon and the PPG-mRNA contents were quantitatively low in comparison to primary rat pancreatic islets. Glucose did not regulate glucagon secretion from INS-1 cells, whereas palmitate induced a threefold increase of glucagon release. From these data, one must conclude
Acknowledgments
The authors acknowledge the excellent technical assistance of Joachim Schweimer. The work was supported by a grant from the Deutsche Forschungsgemeinschaft (DFG) (Bo 1379/2-2).
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Abbreviations: IRI, immunoreactive insulin; PPI, preproinsulin; PPG, preproglucagon.