Amplification and Cloning of Receptor Genes

To amplify Arc/Arg3.1, we used the following primers: for: 5′-CGA AGT GTC CAA GCA GGT G-3′; and rev: 5′-TGA TGG CAT AGG GGC TAA CA-3′. To amplify NO-GC, we used the following primers: for: 5′-ATC CTC TTC AGC GGC ATT GTG-3′ and rev: 5′-TGC ATT GGT TCC TTC TTG CCC-3′. To amplify GC-A, we used the following primers: for: 5′-TGT GAA ACG TGT GAA CCG GA-3′ and rev: 5′-AGG CGG ATC GTT GAA AGG G-3′. To amplify GR, we used the following primers: for: 5′-TCC CCC TGG TAG AGA CGA AG-3′ and rev: 5′-GGC TGG TCG ACC TAT TGA GG-3′. To amplify MR, we used the following primers: for: 5′-GAG ATG AGG CTT CTG GGT GT-3′ and rev: 5′-CAG GAT CAT GGA CGG GGA TG-3′. These fragments were cloned into the pCR II Topo vector (Invitrogen, Karlsruhe, Germany) and their nucleotide sequences were verified by an automated sequencer. Plasmids were isolated using QIAprep Spin Miniprep Kit from Qiagen (Hilden, Germany). Complementary strands for sense and antisense riboprobes were transcribed from either Sp6 or T7 RNA polymerases and labeled using rNTP mix containing digoxigenin labeled uridine triphosphates. All restriction enzymes, RNA polymerases and digoxigenin-labeled rNTP were purchased from Roche Diagnostics (Mannheim, Germany).