Coulochem 3 detector

Male and female rats were pretreated with vehicle or reserpine (1 or 5 mg/kg) as described above. On PD 21, PD 41, and PD 81, male and female preweanling, adolescent, and adult rats (n = 6 male and female rats per group at each age) were decapitated and dorsal striatal sections were dissected bilaterally on an ice-cold dissection plate and stored at −80 °C.
To measure DA and 5-HT content, frozen striatal sections were sonicated in 10 volumes of 0.1 N HClO4 and centrifuged at 20, 000 × g for 20 min at 4 °C. The supernatant was then filtered through a 0.22 μm centrifugation unit at 2, 000 × g for 5 min at 4 °C. Twenty microliters of the resulting extract was assayed for DA and 5-HT using high performance liquid chromatography (HPLC) with electrochemical detection [Hypersil ODS column (150 × 3 mm) and Coulochem III detector; Thermo Fisher Scientific, Waltham, MA]. The mobile phase consisted of 75 mM NaH2PO4, 1.4 mM 1-octane sulfonic acid, 10 mM EDTA, and 10% acetonitrile (pH 3.1) and was pumped at a rate of 0.5 ml/min.

Frozen dorsal striatal and hippocampal samples were sonicated in 200 μl of acetonitrile with 30 μl desipramine hydrochloride (300 ng) and 30 μl (−)-cocaine hydrochloride (3 μg) added as internal standards. Tissue was centrifuged at 20,000 × g for 15 min at 4 °C. Twenty microliters of the resulting extracts were assayed for DA using high-performance liquid chromatography (HPLC) with electrochemical detection [Hypersil ODS column (150 × 3 mm) and Coulochem III detector; Thermo Fisher Scientific, Waltham, MA, USA]. The mobile phase consisted of 75 mM NaH₂PO₄, 1.4 mM 1-octane sulfonic acid, 10 mM EDTA, and 10% acetonitrile (pH 3.1) and was pumped at a rate of 0.5 ml/min.