Glucagon production of the rat insulinoma cell line INS-1—A quantitative comparison with primary rat pancreatic islets

doi:10.1016/j.bbrc.2005.01.168

Biochemical and Biophysical Research Communications

Volume 330, Issue 1, 29 April 2005, Pages 327-332

https://doi.org/10.1016/j.bbrc.2005.01.168 Get rights and content

Abstract

The rat insulinoma cell line INS-1 is the most commonly used clonal cell model in pancreatic β-cell research. Considering the multihormonality of many insulinomas we examined as to how INS-1 cells comply with the notion of resembling a pure β-cell line. Glucagon immunoassays revealed that INS-1 cells secrete glucagon in a similar range as islets. By immunohistochemistry we detected a cytoplasmic glucagon signal in INS-1 cells which colocalized with C-peptide. Cellular content of preproglucagon-mRNA and glucagon protein in INS-1 cells was less than two percent of the respective values in islets, which probably reflects differences in the intracellular metabolism and/or secretory pathways. Taken together, it is obvious that INS-1 cells do not represent an exclusively insulin producing β-cell line.

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.

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Glucagon production of the rat insulinoma cell line INS-1—A quantitative comparison with primary rat pancreatic islets☆

Abstract

Section snippets

Materials and methods

INS-1 cells release both insulin and glucagon in relevant amounts

Conclusions

Acknowledgments

Biochem. Pharmacol.

Pathol. Res. Pract.

J. Biol. Chem.

Clin. Endocrinol. Metab

Metabolism

Metabolism

Semin. Cell Dev. Biol.

J. Biol. Chem.

Establishment of 2-mercaptoethanol-dependent differentiated insulin-secreting cell lines

Endocrinology

While tinkering with the β-cell…metabolic regulatory mechanisms and new therapeutic strategies: American Diabetes Association Lilly Lecture, 2001

Diabetes

Isolation of INS-1-derived cell lines with robust ATP-sensitive K+ channel-dependent and -independent glucose-stimulated insulin secretion

Diabetes

Glucose sensitivity and metabolism-secretion coupling studied during two-year continuous culture in INS-1E insulinoma cells

Endocrinology

Isolation of INS-1-derived cell lines with robust ATP-sensitive K⁺ channel-dependent and -independent glucose-stimulated insulin secretion