Hypothalamic ventromedial COUP-TFII protects against hypoglycemia-associated autonomic failure.

The nuclear receptor Chicken Ovalbumin Upstream Promoter-Transcription Factor II (COUP-TFII) is an important coordinator of glucose homeostasis through its function in different organs such as the endocrine pancreas, adipose tissue, skeletal muscle, and liver. Recently we have demonstrated that COUP-TFII expression in the hypothalamus is restricted to a subpopulation of neurons expressing the steroidogenic factor 1 transcription factor, known to play a crucial role in glucose homeostasis. To understand the functional significance of COUP-TFII expression in the steroidogenic factor 1 neurons, we generated hypothalamic ventromedial nucleus-specific COUP-TFII KO mice using the cyclization recombination/locus of X-overP1 technology. The heterozygous mutant mice display insulin hypersensitivity and a leaner phenotype associated with increased energy expenditure and similar food intake. These mutant mice also present a defective counterregulation to hypoglycemia with altered glucagon secretion. Moreover, the mutant mice are more likely to develop hypoglycemia-associated autonomic failure in response to recurrent hypoglycemic or glucopenic events. Therefore, COUP-TFII expression levels in the ventromedial nucleus are keys in the ability to resist the onset of hypoglycemia-associated autonomic failure.

Interplay among cGMP, cAMP, and Ca2+ in living olfactory sensory neurons in vitro and in vivo.

The mechanism of cGMP production in olfactory sensory neurons (OSNs) is poorly understood, although this messenger takes part in several key processes such as adaptation, neuronal development, and long-term cellular responses to odorant stimulation. Many aspects of the regulation of cGMP in OSNs are still unknown or highly controversial, such as its subcellular heterogeneity, mechanism of coupling to odorant receptors and downstream targets. Here, we have investigated the dynamics and the intracellular distribution of cGMP in living rat OSNs in culture transfected with a genetically encoded sensor for cGMP. We demonstrate that OSNs treated with pharmacological stimuli able to activate membrane or soluble guanylyl cyclase (sGC) presented an increase in cGMP in the entire neuron, from cilia-dendrite to the axon terminus-growth cone. Upon odorant stimulation, a rise in cGMP was again found in the entire neuron, including the axon terminus, where it is locally synthesized. The odorant-dependent rise in cGMP is due to sGC activation by nitric oxide (NO) and requires an increase of cAMP. The link between cAMP and NO synthase appears to be the rise in cytosolic Ca(2+) concentration elicited by either plasma membrane Ca(2+) channel activation or Ca(2+) mobilization from stores via the guanine nucleotide exchange factor Epac. Finally, we show that a cGMP rise can elicit both in vitro and in vivo the phosphorylation of nuclear CREB, suggesting that this signaling pathway may be relevant for both local events (pathfinding, neurotransmitter release) and more distal processes involving gene expression regulation.

Odorant receptors at the growth cone are coupled to localized cAMP and Ca2+ increases.

A distinctive feature in the topographic organization of the olfactory system in mammals is the dual function of the odorant receptor (OR): it detects odors in the nasal epithelium and plays an instructive role in the axonal convergence of olfactory sensory neurons (OSN) into the olfactory bulb (OB). The latter function is supported by genetic experiments and by the expression of the OR not only on the cilia, but also on the axon termini of the OSN. The signaling pathway coupled to the OR on the cilia is well known and is recognized to involve cAMP and Ca(2+), whereas, until now, nothing was known on the functional characteristics of the OR on the axon termini-growth cone. Here, by analyzing the spatiotemporal dynamics of cAMP and Ca(2+) in living OSN in vitro and in situ, we found that the OR at the growth cone is capable of binding odors and is coupled to cAMP synthesis and Ca(2+) influx through cyclic nucleotide gated (CNG) channels. Furthermore we found that selective odor activation of the OR on the growth cone is followed by nuclear translocation of protein kinase A catalytic subunit. These results define the functional properties of the OR on the growth cone and suggest a potential role of OR activation in axonal convergence and sensory map formation.