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Figure 7. Alterations in CRF2A receptor mRNA expression and receptor binding as a result of chronic alprazolam treatment and after spontaneous alprazolam withdrawal. Each bar represents mean ± SEM; n = 6 per group. A, CRF2A receptor mRNA in the LS is increased as a result of chronic alprazolam treatment and subsequently declines to control levels after alprazolam withdrawal. CRF2A receptor mRNA in the VMH is significantly increased at 48 hr after spontaneous alprazolam withdrawal. CRF2A receptor mRNA expression was measured via densitometric analysis subsequent to in situ hybridization performed in 20 µm brain slices from male Sprague Dawley rats after 14 d of a control liquid diet, an alprazolam-supplemented diet at a dose of 90 mg · kg-1 · d-1, and 24, 48, or 96 hr after withdrawal of alprazolam from the diet. +p < 0.05 versus controls by two-tailed t test. *p < 0.05 versus alprazolam; **p < 0.01 versus alprazolam and 24 hr withdrawal by one-way ANOVA followed by Student-Newman-Keuls post hoc pairwise analysis. Representative images of CRF2A mRNA in the LS after in situ hybridization are included as an inset to the graph. B, CRF2 receptor binding decreases in the VMH after spontaneous alprazolam withdrawal. CRF2 receptor binding was measured via densitometric analysis subsequent to autoradiography performed in 20 µm brain slices from male Sprague Dawley rats after 14 d of a control liquid diet, an alprazolam-supplemented diet at a dose of 90 mg · kg-1 · d-1, and 24, 48, or 96 hr after withdrawal of alprazolam from the diet. *p < 0.05 versus all withdrawal time points by one-way ANOVA followed by Student-Newman-Keuls post hoc pairwise analysis. Representative images of CRF2 receptor binding in the VMH after receptor autoradiography are included as an inset to the graph. Because the autoradiographic images after in situ hybridization and receptor autoradiography appear fairly similar, only the region that showed the more statistically significant changes was displayed as an inset for A and B.
