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Blood, 15 April 2007, Vol. 109, No. 8, pp. 3260-3269.
Prepublished online as a Blood First Edition Paper on December 27, 2006; DOI 10.1182/blood-2006-07-036269.


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Submitted July 19, 2006
Accepted December 9, 2006

Comparative gene expression profiling of in vitro differentiated megakaryocytes and erythroblasts identifies novel activatory and inhibitory platelet membrane proteins

I C Macaulay, M R Tijssen, D C Thijssen-Timmer, A Gusnanto, M Steward, P Burns, C F Langford, P Ellis, Frank Dudbridge, J J Zwaginga, N A Watkins*, C. E van der Schoot, and W H Ouwehand

Department of Haematology, University of Cambridge and National Blood Service, Cambridge, United Kingdom
Department of Experimental Immunohaematology, Sanquin Research at CLB, and Landsteiner Laboratory, AMC, University of Amsterdam, Amsterdam, Netherlands
MRC Biostatistics Unit, Institute of Public Health, Cambridge, United Kingdom
Domantis Limited, Cambridge, United Kingdom
Microarray Facility, Wellcome Trust Sanger Institute, Cambridge, United Kingdom
Department of Immunohematology-Blood transfusion, Leiden University Medical Centre, Leiden, Netherlands
Department of Hematology, Amsterdam Medical Centre, University of Amsterdam, Amsterdam, Netherlands

* Corresponding author; email: naw23{at}cam.ac.uk.

To identify previously unknown platelet receptors we compared the transcriptomes of in vitro differentiated megakaryocytes (MKs) and erythroblasts (EBs). RNA was obtained from purified, biologically paired MK and EB cultures and compared using cDNA microarrays. Bioinformatical analysis of MK upregulated genes identified 151 transcripts encoding transmembrane domain containing proteins. Whilst many of these were known platelet genes, a number of previously unidentified or poorly characterized, transcripts were also detected. Many of these transcripts, including G6b, G6f, LRRC32, LAT2 and the G-protein coupled receptor SUCNR1, encode proteins with structural features or functions that suggest they may be involved in the modulation of platelet function. Immunoblotting on platelets confirmed the presence of the encoded proteins, and flow cytometric analysis confirmed the expression of G6b, G6f and LRRC32 on the surface of platelets. Through comparative analysis of expression in platelets and other blood cells we demonstrated that G6b, G6f and LRRC32 are restricted to the platelet lineage, whereas LAT2 and SUCNR1 were also detected in other blood cells. The identification of the succinate receptor SUCNR1 in platelets is of particular interest, as physiologically relevant concentrations of succinate were shown to potentiate the effect of low doses of a variety of platelet agonists.


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