Figure 1.

Intravenous 2-DG and insulin challenges trigger elevations in phospho-ERK1/2 in CRH neurons of the PVH. A1–A7 show single, unprojected confocal images of the PVH in the coronal plane from animals anesthetized and perfused ∼5 min after receiving only anesthesia (control; A4) or 250 mg/kg 2-DG (A1–A3, A5–A7). Similarly, B1–B7 are PVH images from animals perfused 30 min after receiving saline (control; B4) or a 2 U/kg bolus of insulin (B1–B3, B5–B7). A1 and A2, and B1 and B2, are single channel images showing CRH immunoreactivity (green) or phospho-ERK1/2 immunoreactivity (red), and the insets in each figure outline areas enlarged in A5 and A6, and B5 and B6, respectively. Colocalization of CRH with phospho-ERK1/2 in the PVH results in yellow-orange signal, as evident in A3 and B3 for the two challenges. The cellular distribution of the colocalized signals is more readily apparent at 40× magnification in A7 and B7, which are enlargements of the outlined boxes over the PVH shown in A3 and B3, respectively. Scale bars: (in B2) A1, A2, B1, B2, 100 μm; (in B3) A3, B3, 100 μm; (in B4) A4, B4, 100 μm; (inB6) A5, A6, B5, B6, 50 μm; (in B7) A7, B7, 50 μm. All molecule designations in the A panels apply to the corresponding B panels. Note that the fields of view captured in control images A4 and B4 are identical with those shown in A1, A2, and B1, B2, respectively. These data are also presented in pseudocolored form (supplemental Fig. S1, available at www.jneurosci.org as supplemental material).