Hairpin inhibitors directed against mmu-miR-598-3p or the nonmammalian miRNA cel-miR-67 were obtained from GE Dharmacon (Lafayette, CO) and injected bilaterally into the BLA (AP: 1.5 mm, ML: ±3.2 mm from bregma and DV: −4.7 mm from the skull) as previously described (Young et al. 2016 (link)). The injection needle was left in place for 5 min after the infusion was complete. Inhibitors were reconstituted in water then prepared with jetPEI transfection reagent (Polyplus Transfection). One micro liter per hemisphere of 400 ng/µL was injected 28 d after FC, animals remained in the home cage for 2 d, and then were tested for either remote fear memory expression, anxiety-like behavior, and/or sacrificed such that mir-598-3p levels could be measured via qPCR.
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Modulating miR-598-3p in Fear Behavior
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Variable analysis
independent variables
- Hairpin inhibitors directed against mmu-miR-598-3p
- Hairpin inhibitors directed against the nonmammalian miRNA cel-miR-67
- Remote fear memory expression
- Anxiety-like behavior
- Mir-598-3p levels measured via qPCR
- Injection site coordinates (AP: 1.5 mm, ML: ±3.2 mm from bregma and DV: −4.7 mm from the skull)
- Injection needle left in place for 5 min after the infusion was complete
- Inhibitors reconstituted in water then prepared with jetPEI transfection reagent (Polyplus Transfection)
- Injection volume of 1 micro liter per hemisphere of 400 ng/µL
- Time between FC and testing (28 d for remote fear memory, 2 d in home cage)
- cel-miR-67 inhibitor
- Not explicitly mentioned
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