Table 1.
Genes that were up- and down-regulated by the deficiency of P2rx7 in the mouse amygdala
| Symbol | Gene ID | Gene name (accession number) | Agilent (FC) | Real-time PCR (RQ) | TaqMan ID |
|---|---|---|---|---|---|
| (a) | |||||
| Adra2a | 11 551 | Adrenergic receptor α2a (NM_007417) | 2.72 | 1.34 ± 0.38 p = 0.0171 | Mm00845383_s1 |
| Adrb1 | 11 554 | Adrenergic receptor β1 (NM_007419) | 4.77 | Mm00431701_s1 | |
| Ak5 | 229 949 | Adenylate kinase 5 (NM_001081277) | 5.71 | Mm00461978_m1 | |
| Aplp2 | 11 804 | Amyloid β (A4) precursor-like protein 2 (NM_009691) | 5.55 | Mm00507819_m1 | |
| Bdnf | 12 064 | Brain derived neurotrophic factor isoform 1 (NM_007540) | 3.89 | Mm01334047_m1 | |
| Casp8 | 12 370 | Caspase 8 (NM_009812) | 9.57 | Mm00802247_m1 | |
| Chkb | 12 651 | Choline kinase β (NM_007692) | 4.160 | Mm00432498_m1 | |
| Cnr1 | 12 801 | Cannabinoid receptor 1 (NM_007726) | 2.081 | 1.31 ± 0.28 p = 0.0087 | Mm01212171_s1 |
| Ddc | 13 195 | Dopa decarboxylase (NM_016672) | 2.051 | 1.58 ± 0.84 p = 0.0249 | Mm00516688_m1 |
| Drd2 | 13 489 | Dopamine receptor 2 (NM_010077) | 2.37 | 2.44 ± 0.47 p = 0.0120 | Mm00438545_m1 |
| Gabarapl1 | 57 436 | GABAA receptor-associated protein-like 1 (NM_020590) | 4.47 | Mm00457880_m1 | |
| Gabra1 | 14 394 | GABAA receptor subunit α1 (NM_010250) | 4.130 | Mm00439046_m1 | |
| Gabra5 | 110 886 | GABAA receptor subunit α5 (NM_176942) | 7.79 | Mm00621092_m1 | |
| Gabrb2 | 14 401 | GABAA receptor subunit β2 (NM_008070) | 2.141 | 2.51 ± 1.12 p = 0.0036 | Mm00549788_s1 |
| Gabrb3 | 14 402 | GABAA receptor subunit β3 (NM_008071) | 2.041 | 1.34 ± 0.55 p < 0.0005 | Mm00433473_m1 |
| Gabrg1 | 14 405 | GABAA receptor subunit γ1 (NM_010252) | 2.55 | 3.38 ± 1.19 p = 0.0005 | Mm00439047_m1 |
| Gabrg2 | 14 406 | GABAA receptor subunit γ2 (NM_008073) | 2.43 | 2.04 ± 0.89 p = 0.0012 | Mm00433489_m1 |
| Gad1 | 14 415 | Glutamic acid decarboxylase 1 (NM_008077) | 2.171 | 1.10 ± 0.06 p = 0.0009 | Mm00725661_s1 |
| Gfra1 | 14 585 | Glial cell line derived neurotrophic factor family receptor α1 (NM_010279) | 2.231 | 1.82 ± 0.54 p = 0.0308 | Mm00833897_m1 |
| Gja1 | 14 609 | Gap junction protein α1 (NM_010288) | 4.097 | Mm00621092_m1 | |
| Gjb6 | 14 623 | Gap junction protein β6 (NM_001010937) | 4.48 | 1.40 ± 0.28 p < 0.0016 | Mm00433661_s1 |
| Gjc1 | 14 615 | Gap junction protein γ1 (NM_008122) | 2.161 | 1.247 ± 0.11 p = 0.0050 | Mm01253027_m1 |
| Gjd2 | 14 617 | Gap junction protein δ2 (NM_010290) | 4.61 | Mm00439121_m1 | |
| Gje1 | 76 743 | Gap junction membrane channel protein ε1 (NM_080450) | 2.00 | 1.49 ± 0.18 p = 0.0022 | Mm00519120_s1 |
| Glra2 | 237 213 | Glycine receptor α2 subunit (NM_183427) | 2.34 | 1.30 ± 0.25 p = 0.0157 | Mm00439140_m1 |
| Glrb | 14 658 | Glycine receptor β subunit (NM_010298) | 5.089 | 1.16 ± 0.25 p = 0.0018 | Mm00439140_m1 |
| Gria2 | 14 800 | Glutamate receptor, ionotropic, AMPA2 (α2) | 2.59 | 1.13 ± 0.05 p < 0.0002 | Mm00442822_m1 |
| Gria4 | 14 802 | Glutamate receptor, ionotropic, AMPA4 (α4), transcript variant 1 (NM_019691) | 2.74 | 1.41 ± 0.66 p = 0.0014 | Mm00444754_m1 |
| Grm7 | 108 073 | Glutamate receptor, metabotropic 7 (NM_177328) | 35.81 | 1.07 ± 0.16 p = 0.0317 | Mm01189424_m1 |
| Gstm4 | 14 865 | Glutathione S-transferase μ4 (NM_026764) | 3.34 | Mm00728197_s1 | |
| Klhl12 | 240 756 | Kelch-like 12 (Drosophila) (NM_153128) | 6.98 | 1.19 ± 0.31 p = 0.0074 | Mm00462323_m1 |
| Narg1l | 66 897 | NMDA receptor regulated 1-like (NM_025832) | 6.98 | 1.77 ± 0.21 p = 0.0032 | Mm00462323_m1 |
| Ncam1 | 17 967 | Neural cell adhesion molecule 1 (NM_010875) | 3.310 | Mm00456815_m1 | |
| Npy5r | 18 168 | Neuropeptide Y receptor Y5 (NM_016708) | 6.68 | Mm02620267_s1 | |
| P2ry1 | 18 441 | Purinergic receptor P2Y, G-protein coupled 1 (NM_008772) | 3.89 | 1.47 ± 0.73 p = 0.0467 | Mm00435471_m1 |
| Pcdh10 | 18 526 | Protocadherin 10 (NM_001098171) | 14.93 | Mm00477987_s1 | |
| Pcmt1 | 18 537 | Protein-l-isoaspartate (d-aspartate) O-methyltransferase 1 (NM_008786) | 2.251 | 1.23 ± 0.28 p = 0.0073 | Mm00476600_m1 |
| Pde1a | 18 573 | Phosphodiesterase 1A calmodulin- dependent (NM_016744) | 2.191 | 1.24 ± 0.74 p = 0.0123 | Mm00450244_m1 |
| Plp1 | 18 823 | Proteolipid protein (myelin) 1 (NM_011123) | 9.031 | 1.51 ± 0.18 p = 0.0033 | Mm00456892_m1 |
| Ryr3 | 20 192 | Ryanodine receptor 3 (BC116740) | 2.80 | 1.79 ± 0.22 p < 0.0002 | Mm01335482_m1 |
| (b) | |||||
| Adcy8 | 11 514 | Adenylate cyclase (NM_009623) | 17.56 | Mm00507722_m1 | |
| Adrbk1 | 110 355 | Adrenergic receptor kinase β1 (NM_130863) | 5.55 | Mm00804778_m1 | |
| Adrbk2 | 320 129 | Adrenergic receptor kinase β2 (NM_177080) | 4.38 | Mm00622037_m1 | |
| Aloxe3 | 23 801 | Arachidonate lipoxygenase 3 (NM_011786) | 5.121 | Mm004778628_m1 | |
| Anxa7 | 11 750 | Annexin A7 (NM_009674) | 3.94 | Mm00477549_m1 | |
| Avp | 11 998 | Arginin vasopressine (NM_009732) | 57.00 | Mm00437761_g1 | |
| Bai1 | 107 831 | Brain-specific angiogenesis inhibitor 1 (NM_174991) | 2.52 | Mm00558144_m1 | |
| Bdrkb1 | 12 061 | Bradykinin receptor β1 (NM_007539) | 27.79 | 1.60 ± 0.43 p = 0.0498 | Mm00432059_s1 |
| Crhr2 | 12 922 | Corticotropin releasing hormone receptor 2 (NM_007539) | 5.110 | 5.89 ± 0.43 p = 0.0040 | Mm00438303_m1 |
| Drd4 | 13 491 | Dopamine receptor 4 (NM_007878) | 4.580 | Mm004382893_m1 | |
| Gabrd | 14 403 | GABAA receptor subunit delta (NM_008072) | 2.72 | Mm00433476_m1 | |
| Gabrr1 | 14 408 | GABAC receptor, subunit rho (NM_008075) | 4.123 | Mm00433499_m1 | |
| Gcat | 26 912 | Glycine C-acetyltransferase (2-amino-3 ketobutyrate-coenzyme A ligase) (NM_013847) | 25.45 | Mm00496962_m1 | |
| Glul | 14 645 | glutamate-ammonia ligase (glutamine synthetase) (NM_008131) | 4.56 | Mm00725701_s1 | |
| Grik1 | 14 805 | Glutamate receptor, ionotropic, kainate 2 (NM_010348) | 12.130 | Mm00446882_m1 | |
| Grik5 | 14 809 | Glutamate receptor, ionotropic, kainate 5 (γ2) (NM_008168) | 9.95 | Mm00435701_m1 | |
| Grin2b | 14 812 | Glutamate receptor, ionotropic, NMDA2B (ε2) (NM_008171) | 2.77 | 1.10 ± 0.13 p = 0.0062 | Mm00433820_m1 |
| Grin2d | 14 814 | Glutamate receptor, ionotropic, NMDA2D (ε4) (NM_008171) | 7.168 | Mm00433822_m1 | |
| Grm1 | 14 816 | Glutamate receptor metabotropic 1 (NM_016976) | 33.64 | Mm00810231_s1 | |
| Grm5 | 108 071 | Glutamate receptor metabotropic 5 (NM_001081414) | 6.110 | Mm00690332_m1 | |
| Nrxn2 | 18 190 | Neurexin II (NM_020253) | 7.131 | 1.28 ± 0.18 p < 0.0328 | Mm01236851_m1 |
| P2rx2 | 231 602 | Purinergic receptor P2X, ligand-gated ion channel, 2 (NM_153400) | 11.180 | Mm00462952_m1 | |
GABA, γ-aminobutyric acid; NMDA, N-Methyl-d-aspartate.
The table shows the comparison of gene expression data from microarray and real-time polymerase chain reaction (PCR) studies. The detected changes in the expression level are expressed in fold change (FC) in the microarray study and relative quantity (RQ) in the PCR study. Among the representative list of 60 genes of interest, 39 were down-regulated (a) and 21 were up regulated (b) exclusively by P2rx7 deletion in the mouse amygdala using the selection filters of >2.0-FC in the microarray study. Significant differences were calculated by a two-way analysis of variance (ANOVA) followed by Benjamini–Hochberg's multiple correction, with p < 0.05. Significant down- or up-regulatory effects of P2rx7 deficiency were confirmed for 29 of the genes by TaqMan-based quantitative real-time PCR (TLDA). Samples of RNA from the microarray experiment (n = 4) and from an independent experiment (n = 4) were measured for each group on TLDA and the results were evaluated by RealTime StatMiner software. (a) Twenty-five genes among the total of 39 were changed significantly (two-way ANOVA, p < 0.05) and showed similarity in size and direction by both methods. (b) Four out of 21 genes were changed significantly (two-way ANOVA, p < 0.05) and showed similarity in size and direction by both methods. The TaqMan ID gives the product number of Applied Biosystems Gene Expression Assay used for the validation of the corresponding gene.