Figure 2.

CART expression in cultured vagal afferent neurons. Vagal afferent neurons cultured for >48 h showing CART immunoreactivity (A, C, E, G, I, K) or nuclei stained with DAPI in the respective fields (B, D, F, H, J, L). A, B, CART is undetectable after transfer of neurons to serum-free medium for 2 h. C, D, CART immunoreactivity is readily detected in cells in serum-free medium 2 h after the addition of 10 nm CCK. E, F, CART is also induced in cells in serum-free medium 2 h after the addition of 100 nm PMA. G, H, The effect of CCK is reduced by the PKC inhibitor Ro-32-0432 (1 μm). I–L, In serum-free medium, ghrelin (10 nm) has no effect on CART abundance (I, J) but inhibits the effect of CCK (K, L). Arrows identify a representative neuron in each pair of images; filled arrows show CART-positive neurons, and open arrows show CART-negative neurons. Representative images from six independent experiments are shown. Scale bar, 50 μm.