Nitric oxide in the nucleus of the tractus solitarius is involved in hypoglycemic conditioned response

The repeated injection of insulin (unconditioned stimulus, UCS) immediately followed by exposure to sensory stimulation (e.g. sound or odor; conditioned stimulus, CS) results in a learned conditioned reflex in which the exposure to the CS alone lowers blood glucose. The brain regions participating in this hypoglycemic Pavlovian response remain unknown. Here we investigate if nitric oxide (NO) in the nucleus tractus solitarius (NTS), a nucleus known to be involved in glucose homeostasis, participates in this hypoglycemic reflex. Insulin injections (UCS) were paired with exposure to menthol odor (CS). After 8-10 reinforcements (4-5days training), rats acquire the learned hypoglycemic response. An increase in c-Fos expression was observed in the NTS, the ventrolateral hypothalamic nucleus (VLH) and other brain regions of conditioned rats. Microinjections of 3-(5'-hydroxymethyl-2'furyl)-1-benzyl indazole (YC-1) a stimulator of soluble guanylate cyclase (sGC) into NTS before the UCS accelerated the acquisition of the learned hypoglycemic response; 5-6 reinforcement produced pronounced glucose drop when exposed to the CS. In contrast, an inhibitor of NO synthase (NOS) N^ω-Nitro-l-arginine methyl ester (L-NAME) in the NTS prolonged the required training period (11-15 reinforcements) to obtain the hypoglycemic reflex, and reduced the glycemic response. The number of c-Fos expressing cells in the NTS and VLH in rats receiving YC-1was significantly higher than that observed in rats receiving L-NAME. These findings suggest that NO-cGMP-PKG signaling in the NTS can modify the acquisition of conditioned hypoglycemia, and suggests that this nucleus directly participates in this reflex.