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Figure 1 | BMC Cell Biology

Figure 1

From: S-Nitrosothiols modulate G protein-coupled receptor signaling in a reversible and highly receptor-specific manner

Figure 1

S-nitrosoglutathione (GSNO) reversibly modulates basal and receptor-dependent G protein activity in rat brain cryostat sections. [35S]GTPγS autoradiography of sagittal brain sections was conducted using a 3-step protocol with DPCPX (10-6 M) present throughout steps 2 and 3, as detailed in the Methods section. Where indicated, GSNO (0.5 mM) was present for 60 min during the GDP loading (step 2). When used, DTT (1 mM) or GSH (1 mM) were present during the [35S]GTPγS labeling (step 3). The muscarinic agonist, carbachol (CCh, 10-4 M), the P2Y receptor agonist 2-methylthio-ADP (2MeSADP, 10-5 M) or lysophosphatidic acid (LPA, 5 × 10-5 M in 0.1% fatty acid free BSA) were present in step 3. In the control panel (left), the anatomical loci where receptor agonists typically activate G proteins are indicated. Note GSNO-dependent overall increase in basal G protein activity, as well as robust amplification of CCh-stimulated G protein activity in several gray matter regions visible at this sagittal plane, most notably the brain stem (bs) nuclei, the striatum (Str), and the superficial gray layer of the superior colliculus (SuG). Note also clear attenuation of 2MeSADP-stimulated responses in all brain regions, and blunting of LPA-stimulated responses, especially in the white matter areas, including the corpus callosum (cc), the fimbria of the hippocampus (fi) and the cerebellar white matter (Cbw). Scale bar = 2 mm. For quantitative data on selected brain regions, see Supplementary Figs. 1 and 2 in additional file 1.

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