Comparative gene expression profiling of in vitro differentiated megakaryocytes and erythroblasts identifies novel activatory and inhibitory platelet membrane proteins.
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-up-regulated genes identified 151 transcripts encoding transmembrane domain-containing proteins. Although 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, because physiologically relevant concentrations of succinate were shown to potentiate the effect of low doses of a variety of platelet agonists.
Macaulay
Iain C
IC
Department of Haematology, University of Cambridge, Cambridge, UK.
Tijssen
Marloes R
MR
Thijssen-Timmer
Daphne C
DC
Gusnanto
Arief
A
Steward
Michael
M
Burns
Philippa
P
Langford
Cordelia F
CF
Ellis
Peter D
PD
Dudbridge
Frank
F
Zwaginga
Jaap-Jan
JJ
Watkins
Nicholas A
NA
van der Schoot
C Ellen
CE
Ouwehand
Willem H
WH
D002454:Cell Differentiation / Q000502:physiology*; D004900:Erythroblasts / Q000166:cytology / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010980:Platelet Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics
D010980:Platelet Membrane Glycoproteins