Sensitive and specific method for detecting G protein–coupled receptor mRNAs

Arne Hansen^1, 3, Yidong Chen², Jason M Inman⁵, Quang N Phan^4, 6, Zhi-Qing Qi⁴, Charlie C Xiang^4, 6, Miklos Palkovits⁴, Natasha Cherman¹, Sergei A Kuznetsov¹, Pamela G Robey¹, Eva Mezey^1, 3 & Michael J Brownstein^4, 6

¹  Craniofacial and Skeletal Diseases Branch, National Institute of Dental and Craniofacial Research, Department of Health and Human Services, Bethesda, Maryland 20892, USA.

²  Cancer Genetics Branch, National Human Genome Research Institute, Department of Health and Human Services, Bethesda, Maryland 20892, USA.

³  Laboratory of Neurochemistry, National Institute of Neurological Disorder and Stroke, Department of Health and Human Services, Bethesda, Maryland 20892, USA.

⁴  National Institute of Mental Health, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland 20892, USA.

⁵  The Institute for Genomic Research, Rockville, Maryland 20850, USA.

⁶  J. Craig Venter Institute, Rockville, Maryland 20850, USA.

G protein–coupled receptors (GPCRs) mediate effects of extracellular signaling molecules in all the body's cells. These receptors are encoded by scarce mRNAs; therefore, detecting their transcripts with conventional microarrays is difficult. We present a method based on multiplex PCR and array detection of amplicons to assay GPCR gene expression with as little as 1 g of total RNA, and using it, we profiled three human bone marrow stromal cell (BMSC) lines.

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