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
- 1Gene Expression Unit, Department of Molecular Cell Biology, Katholieke Universiteit Leuven, Leuven, Belgium
- 2Cell Pharmacology Unit, Department of Molecular Cell Biology, Katholieke Universiteit Leuven, Leuven, Belgium
- 3Unit of Endocrinology and Metabolism, Faculty of Medicine, Université Catholique de Louvain, Brussels, Belgium
- 4Diabetes 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
Abstract
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 β-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.
- AUC, area under the curve
- DMEM, Dulbecco’s modified Eagle’s medium
- FACS, fluorescence-activated cell sorter
Footnotes
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Additional information for this article can be found in an online appendix at http://diabetes.diabetesjournals.org.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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- Accepted September 14, 2006.
- Received June 7, 2006.
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