Table 1.
Forward (F) and reverse (R) primers used for RT-PCR analysis of GABAergic marker expressiona
| Marker | Accession number | Primer sequences | Annealing position | Product size (bp) | Reference |
|---|---|---|---|---|---|
| Receptor subunits | |||||
| α1 | NM_183326 | F 5′-AGCTATACCCCTAACTTAGCCAGG-3′ | 1341–1364 | 304 | Hayasaki et al. 2006 |
| R 5′-AGAAAGCGATTCTCAGTGCAGAGG-3′ | 1645–1622 | ||||
| α2 | L08491 | F 5′-ACAACGGGAAAAAGTCAGTG-3′ | 329–348 | 282 | Stewart et al. 2002 |
| R 5′-TGGCCCAGCAAATCATAC-3′ | 661–594 | ||||
| β2 | X15467 | F 5′-GAGAAGATGCGCCTGGATGTCA-3′ | 1127–1148 | 389 | Stewart et al. 2002 |
| R 5′-ATGGGAGGCTGGAGTTTAGTTCAC-3′ | 1516–1453 | ||||
| γ2S | L08497 | F 5′-AAGAAAAACCCTGCCCCTACCATT-3′ | 1158–1181 | 336 | Stewart et al. 2002 |
| R 5′-TTCGTGAGATTCAGCGAATAAGAC-3′ | 1494–1471 | ||||
| γ2L | BC092617 | F 5′-CTTCTTCGGATGTTTTCCTTCAAG-3′ | 389–412 | 390 | Stewart et al. 2002 |
| R 5′-CATAGGGTATTAGATCGTTGGACT-3′ | 779–756 | ||||
| γ3 | NM_024370 | F 5′-CATCCAGATTCAACAAGATG-3′ | 1189–1208 | 253 | Hayasaki et al. 2006 |
| R 5′-AGCTCAGAGACGTCAATG-3′ | 1442–1425 | ||||
| δ | NM_017289 | F 5′-TGAGGAACGCCATTGTCCTCTTCT-3′ | 1116–1139 | 333 | Hayasaki et al. 2006 |
| R 5′-ACCACCGCACGTGGTACATGTAAA-3′ | 1449–1426 | ||||
| Transporter isoforms | |||||
| GAT 1 | NM_024371 | F 5′-ACGCTTCGACTTCCTCATGTCCTGT-3′ | 296–320 | 698 | Fujimori et al. 2006 |
| R 5′-GAATCAGACAGCTTTCGGAAGTTGG-3′ | 994–970 | ||||
| GAT 2 | NM_133623 | F 5′-CCGGGTGTTCCGTAAGAAGAACCG-3′ | 1346–1369 | 541 | Howd et al. 1997 |
| R 5′-GTGTCGCTGGAGAAGTCAGCTCTG-3′ | 1887–1864 | ||||
| GAT 3 | NM_024372 | F 5′-ACCCCAAGGCTGTCACTATG-3′ | 1142–1161 | 479 | Fujimori et al. 2006 |
| R 5′-TGTTGTACTTGAGCGGCTTG-3′ | 1621–1602 | ||||
| BGT-1 | NM_017335 | F 5′-AGGGAGGCGTTCACCTCAGG-3′ | 2092–2111 | 388 | Fujimori et al. 2006 |
| R 5′-TTGGGCTCTGCAAGGCTGC-3′ | 2480–2462 | ||||
| Biosynthesis enzyme isoforms | |||||
| GAD 65 | NM_012563 | F 5′-GGCTCTGGCTTTTGGTCCTTC-3′ | 87–107 | 437 | Hayasaki et al. 2006 |
| R 5′-TGCCAATTCCCAATTATACTCTTGA-3′ | 524–500 | ||||
| GAD 67A | NM_017007 | F 5′-GCTGGAAGGCATGGAAGGTTTTAA-3′ | 621–644 | 302 | Hayasaki et al. 2006 |
| R 5′-AATATCCCATCACCATCTTTATTTGACC-3′ | 923–896 | ||||
| GAD 67E | M383510 | F 5′-GCTGGAAGGCATGGAAGGTTTTA-3′ | 538–560 | 243 | Szabo et al. 2000 |
| R 5′-TGAGCCCCATCACCGTAGCA-3′ | 781–762 | ||||
| Control housekeeping | |||||
| Lamin | NM_001002016 | F 5′-GCAGTACAAGAAGGAGCTA-3′ | 933–951 | 318 | Brown & Nurse, 2008 |
| R 5′-CAGCAATTCCTGGTACTCA-3′ | 1251–1233 | ||||
a
All primers were synthesized by MOBIX (McMaster University, Hamilton, Canada). Annealing temperature during amplification was 55°C, except for GAT 1 (60°C), GAT 3 (60°C) and BGT-1 (65°C).