We have shown earlier that repeated stress increases 2-AG in the amygdaloid region, an effect that shows sensitization such that maximal increases are seen after 10 consecutive days of restraint (Patel and Hillard, 2008; Rademacher et al, 2008). We sought to extend these findings by increasing our anatomical resolution to examine the BLA subregion of the amygdala, and determine the effects of repeated restraint stress on 2-AG precursors and metabolites in this region. We developed an analytical approach that allowed for the simultaneous detection of 2-AG, AA, OAG, and SAG from 1 mm micropunches of the BLA (mean tissue weight 5.0 ± 0.2 mg, N = 18; punch location shown in Figure 2A and B). A representative chromatogram depicting retention times and high signal-to-noise ratio for the endogenous compounds, and corresponding deuterated internal standards, are shown in Figure 2C. We stressed mice for either 20 min or 1 h, or for 1 h per day for 10 consecutive days. Chronically stressed mice gained significantly less weight over the 10-day restraint protocol than control mice (control 4.4 ± 0.4g N =
