To identify the effects of intra-NAc infusions of β-endorphin on ethanol tremor, anxiety-like behavior, ethanol self-administration, and levels of TH immunoreactivity in ethanol-dependent rats, bilateral guide cannulae (Plastic One, Roanoke, VA, USA) aimed at the NAc were implanted under anesthesia stereotaxically using the following coordinates: NAc shell region (AP, 1.7; ML, ±0.75; and DV, 5.7 from bregma). The atlas of Paxinos and Watson (42) was used as a reference. After 7 days of recovery, rats were fed by an ethanol-containing diet using the same ethanol diet schedule and then withdrawn from ethanol for 2 hours. Following a 2-hour ethanol withdrawal, β-endorphin (0.25 μg/0.5 μl per side) was injected bilaterally into the NAc more than 1 min with a 33-gauge injection needle extending 1 mm beyond the guide cannulae tip. The injectors were left in place for 2 min to allow diffusion into brain tissues before removal. β-Endorphin was dissolved in 0.9% sterile saline. An equal volume of saline was injected into the NAc for comparative purposes.

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