Riboprobes for GluA1, Homer 1 and PSD-95 (Supplement 1) were generated by in vitro transcription using SP6 or T7 polymerase and [35S]-αUTP (Amersham Biosciences). In situ hybridization was performed on 20μm-thick cryosections from the freshly frozen amygdala samples as previously described (23, 24)(information provided in Supplement 1). Briefly, brain sections were incubated with 270 ml (20×103 CPM/ml) overnight at 55° C and following post-hybridization washes were exposed to Kodak Biomax MR film for 5–15 days.
