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Because LC3 is a reliable marker of autophagosomes, LC3-GFP mice are widely used to monitor autophagy in?vivo (Mizushima et?al., 2004). LC3-GFP puncta were readily detectable in various regions of the CNS in basal conditions (Figure?S1), including in the ARH (Figure?1A). LC3-GFP puncta were found in the ARH as early as postnatal day 10, and the presence of autophagosomes persisted at weaning and into adulthood (Figure?1A). The density of LC3-GFP puncta displayed no significant variations across the ages studied (Figure?1A), and puncta were detected in both perikaryons and processes (Figure?1A). The LC3-GFP signal was not restricted to the ARH; it was also observed in processes located in the periventricular zone of the hypothalamus, which appears to be the major route for ascending ARH efferent connections (Bouret et?al., 2004a?and?Bouret et?al., 2004b) (Figure?1A). The [http://www.selleckchem.com/products/ulixertinib-bvd-523-vrt752271.html selleck compound] presence of constitutive autophagy in the hypothalamus was further confirmed by the presence of both LC3 I (soluble form) and LC3 II (membrane-bound form) immunoreactivity in the adult hypothalamus under basal conditions (Figure?1B). Because POMC is expressed in ARH neurons and in axon terminals traveling throughout the periventricular zone of the hypothalamus (Bouret et?al., 2004a), i.e., where basal autophagy was detected, we next investigated whether autophagy occurs specifically in POMC neurons by using electron microscopy. Ultrastructural analysis of material derived from P24 wild-type mice revealed the presence of double-membraned autophagosomes in [http://www.selleckchem.com/products/Temsirolimus.html http://www.selleckchem.com/products/Temsirolimus.html] both neuronal perikarya and neuronal processes of POMC neurons, including dendrites (Figure?1C). Together, these data indicate that autophagy occurs constitutively in the hypothalamus. The presence of autophagosomes specifically in POMC neuronal processes suggests a role for hypothalamic autophagy in metabolic regulation. To determine the physiological role of autophagy in POMC neurons, we generated mice that lack Atg7, an essential autophagy gene, specifically in POMC neurons. We crossed mice [http://en.wikipedia.org/wiki/OPHN1 OPHN1] carrying an Atg7loxP allele ( Komatsu et?al., 2005) with mice that express Cre recombinase in a POMC-specific manner (Pomc-Cre) ( Balthasar et?al., 2004). The resulting Pomc-Cre; Atg7loxP/loxP mice were born normally and survived to adulthood. Pomc-Cre; Atg7loxP/loxP mice had body weights undistinguishable from their Atg7loxP/loxP control littermates until 6?weeks of age (group, F(1/322)?= 2.08, p?= 0.1498; age, F(9/322)?= 159.22, p?