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Diabetes 55:3214-3220, 2006
DOI: 10.2337/db06-0774
© 2006 by the American Diabetes Association
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Probe-Independent and Direct Quantification of Insulin mRNA and Growth Hormone mRNA in Enriched Cell Preparations

Leentje Van Lommel1, Kristel Janssens2, Roel Quintens1, Katsura Tsukamoto1, Dirk Vander Mierde1, Katleen Lemaire1, Carl Denef2, Jean-Christophe Jonas3, Geert Martens4, Daniel Pipeleers4, and Frans C. Schuit1

1 Gene Expression Unit, Department of Molecular Cell Biology, Katholieke Universiteit Leuven, Leuven, Belgium
2 Cell Pharmacology Unit, Department of Molecular Cell Biology, Katholieke Universiteit Leuven, Leuven, Belgium
3 Unit of Endocrinology and Metabolism, Faculty of Medicine, Université Catholique de Louvain, Brussels, Belgium
4 Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium

Address correspondence and reprint requests to Frans C. Schuit, Herestraat 49, P.O. Box 901, B-3000 Leuven, Belgium. E-mail: frans.schuit{at}med.kuleuven.be

Abbreviations: AUC, area under the curve; DMEM, Dulbecco’s modified Eagle’s medium; FACS, fluorescence-activated cell sorter

Task division in multicellular organisms ensures that differentiated cell types produce cell-specific proteins that fulfill tasks for the whole organism. In some cases, the encoded mRNA species is so abundant that it represents a sizeable fraction of total mRNA in the cell. In this study, we have used a probe- and primer-free technique to quantify such abundant mRNA species in order to assess regulatory effects of in vitro and in vivo conditions. As a first example, we were able to quantify the regulation of proinsulin mRNA abundance in ß-cells by food intake or by the glucose concentration in tissue culture. The second example of application of this technique is the effect of corticosteroids on growth hormone mRNA in enriched somatrotrophs. It is anticipated that other examples exist in which measurement of very abundant mRNAs in dedicated cells will help to understand biological processes, monitor disease states, or assist biotechnological manufacturing procedures.


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Copyright © 2006 by the American Diabetes Association.