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Increased Cytokine-Induced Cytotoxicity of Pancreatic Islet Cells from Transgenic Mice Expressing the Src-like Tyrosine Kinase GTK

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Abstract

Background

The loss of β cells in type 1 diabetes may involve protein kinases because they control cell growth, differentiation, and survival. Previous studies have revealed that GTK, a Src-like protein tyrosine kinase expressed in β cells (also named Bsk/Iyk), regulates multiple responses including growth and survival of rat insulinoma cells (RINm5F) and differentiation of neuronal PC12 cells. In the present study, we have generated a transgenic mouse expressing a kinase active GTK mutant (GTK-Y504F) under the control of the rat insulin I promoter to establish a role of GTK in β cells.

Materials and Methods

Control and GTK-transgenic CBA mice were used for determination of in vivo glucose tolerance and the relative insulin-positive area. Isolated islets from both groups were cultured in the absence and presence of cytokines and insulin secretion, viability and protein expression were assessed.

Results

The β-cell mass of GTK-transgenic mice was increased as a consequence of a larger pancreas and an increased relative β-cell area. Islets isolated from the transgenic animals exhibited an enhanced glucose-induced insulin release and reduced viability in response to cytokines that could not be explained by higher levels of nitric oxide (NO) compared with control islets. Extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen-activated protein kinase (MAPK), c-Jun NH2-terminal kinase (JNK), and Akt were all activated by cytokines, but GTK-transgenic islets contained higher basal levels of phosphorylated ERK1/2 and lower basal levels of phosphorylated p38 compared with the control islets. The total amount of activated MAPKs was, however, higher in the cytokine-stimulated transgenic islets compared with the control islets due to increased levels of phospho-ERK1/2. Moreover, the proline-rich tyrosine kinase (PYK) 2 (also named RAFTK/CAK β/CADTK) levels were elevated in response to a 24-hr exposure to cytokines in control islets but not in the GTK-transgenic islets.

Conclusions

These results suggest that although GTK increases the β-cell mass, it also enhances islet cell death in response to cytokines and may thus be involved in the β-cell damage in type 1 diabetes.

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Acknowledgments

We are grateful to Drs Helena Edlund and Ulf Ahlgren for help with generating the transgenic mice and to Ing-Britt Hallgren and Ing-Marie Mörsare for technical assistance. This work was supported by grants from the Juvenile Diabetes Foundation International, the Swedish Medical Research Council (31X-10822), the Swedish Diabetes Association, the Novo-Nordisk Foundation, and the Family Ernfors Fund.

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  1. Department of Medical Cell Biology, Uppsala University, Box 571, Biomedicum, S-751 23, Uppsala, Sweden

    Cecilia Annerén & Michael Welsh

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  1. Cecilia Annerén
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  2. Michael Welsh
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Correspondence to Cecilia Annerén.

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Contributed by D.F. Steiner.

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Annerén, C., Welsh, M. Increased Cytokine-Induced Cytotoxicity of Pancreatic Islet Cells from Transgenic Mice Expressing the Src-like Tyrosine Kinase GTK. Mol Med 7, 301–310 (2001). https://doi.org/10.1007/BF03402213

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