Table 3.
G-coupled protein receptors present in corporal smooth muscle cells (detected by microarray analysis) and change in expression when treated with Vcsa1-NEP.
| Symbol | Gene | B-statistic | Fold Change |
|---|---|---|---|
| Gabra4 | gamma-aminobutyric acid (GABA-A) receptor, subunit alpha 4 | 12.41037488 | 20.68691607 |
| Ccr2 | chemokine (C-C motif) receptor 2 | 14.20093557 | 17.19107659 |
| Hcrtr2 | hypocretin (orexin) receptor 2 | 12.51012791 | 15.86347807 |
| Mtnr1a | melatonin receptor 1A | 11.43199341 | 11.21879566 |
| Celsr3 | cadherin EGF LAG seven-pass G-type receptor 3 | 9.0069665 | 10.29157574 |
| Ccr5 | chemokine (C-C) receptor 5 | 10.67853803 | 9.966672355 |
| Agtr2 | angiotensin II receptor, type 2 | 7.787873351 | 9.154239457 |
| Adcyap1r1 | adenylate cyclase activating polypeptide 1 receptor 1 | 11.69137911 | 8.768922888 |
| Oprm1 | opioid receptor, mu 1 | 9.947061929 | 7.321373484 |
| Grm2 | glutamate receptor, metabotropic 2 | 9.432481922 | 6.876953784 |
| Calcrl | calcitonin receptor-like | 10.07497323 | 6.42708125 |
| Fshr | follicle stimulating hormone receptor | 8.764853382 | 6.34367168 |
| Nxph1 | neurexophilin 1 | 9.195471437 | 5.415030946 |
| Arl3 | ADP-ribosylation factor-like 3 | 8.571346447 | 4.960725826 |
| Gpr24 | G protein-coupled receptor 24 | 5.88419255 | 4.737365123 |
| Ecel1 | endothelin converting enzyme-like 1 | 6.804210975 | 4.468540643 |
| C3ar1 | complement component 3a receptor 1 | 8.560846324 | 4.408652995 |
| Gabrr2 | gamma-aminobutyric acid A receptor, rho 2 | 8.652629815 | 4.22493427 |
| Chrm5 | cholinergic receptor, muscarinic 5 | 3.675843876 | 3.866131237 |
| Gabrg2 | gamma-aminobutyric acid A receptor, gamma 2 | 7.981849786 | 3.491850763 |
| Mas1 | MAS1 oncogene | 6.703196086 | 2.915382323 |
| Grb14 | growth factor receptor bound protein 14 | 0.279054051 | 2.846778548 |
| Sstr1 | somatostatin receptor 1 | 5.813340243 | 2.811720785 |
| Vipr1 | vasoactive intestinal peptide receptor 1 | 5.854036807 | 2.573352521 |
| Ghrh | growth hormone releasing hormone | 3.464521913 | 2.516117866 |
| Grik2 | glutamate receptor, ionotropic, kainate 2 | 4.558269023 | 2.430048197 |
| Drd2 | dopamine receptor 2 | 6.328035066 | 2.415420604 |
| Adrb2 | adrenergic receptor, beta 2 | 4.83859972 | 2.291419935 |
| F2r | coagulation factor II (thrombin) receptor | 4.981961453 | 1.888623241 |
| Htr5a | 5-hydroxytryptamine (serotonin) receptor 5A | 5.022532023 | 1.801031313 |
| Chrm2 | cholinergic receptor, muscarinic 2 | 1.017576296 | 1.446614704 |
| Gria3 | glutamate receptor, ionotropic, AMPA3 (alpha 3) | 3.123862632 | 1.44637896 |
| Gpr20 | G protein-coupled receptor 20 | 1.776954221 | 1.410922562 |
| Chrm3 | cholinergic receptor, muscarinic 3 | 3.348453846 | 1.339036933 |
| Htr7 | 5-hydroxytryptamine (serotonin) receptor 7 | 0.47710741 | 1.18283939 |
| Ptgdr | prostaglandin D receptor | 0.239210351 | 0.998857706 |
| Adrb3 | adrenergic receptor, beta 3 | 1.805930858 | 0.984280774 |
| Drd4 | dopamine receptor 4 | 2.043147208 | 0.949623286 |
| Tacr2 | tachykinin receptor 2 | 0.981167624 | 0.931291478 |
| P2ry2 | purinergic receptor P2Y, G-protein coupled 2 | 0.604120257 | 0.882479154 |
| Gng8 | guanine nucleotide binding protein (G protein), gamma 8 subunit | 1.150735789 | 0.813719316 |
| Nos2 | nitric oxide synthase 2, inducible | 0.134838156 | 0.704032284 |
| Gnas | GNAS complex locus | 0.461991262 | 0.632688694 |
A complete list of the G-coupled protein receptors with changed expression in following knock-down of expression of Vcsa1 using siRNA. RNA was prepared from treated and untreated cells, reverse transcribed to cDNA, labeled with biotin and fragmented according to protocols developed by Affymetrix (www.affymetrix.com). Subsequent hybridization to microarray chips was performed at AECOM microarray core facilities using an Affymetrix Fluidics Station. Analysis of the data was performed AffylmGUI, available from www.bioconductor.org. Briefly the steps were to visually check for artifacts on the Chip (and exclude outlying chips from further analysis), normalize raw data using robust multi-array averaging (RMA) and calculate the fold change in level of gene expression by comparing experiment and control groups. Significant change was determined by the B statistic (which is the log odds of differential expression, which takes into account the variability between replicate arrays).