Prolyl carboxypeptidase in Agouti-related Peptide neurons modulates food intake and body weight

Highlights

• PRCP is expressed in the arcuate NPY/AgRP neurons.

• PRCP is released in the synaptic space following ghrelin stimulation.

• Male mice with PRCP deletion in NPY/AgRP neurons show leaner phenotype with decreased food intake on standard chow diet.

• Female mice with PRCP deletion in NPY/AgRP neurons show leaner phenotype with decreased food intake only on high fat diet.

• Selective blockade of PVN melanocortin receptors increases feeding in male mice with PRCP deletion in NPY/AgRP neurons.

Abstract

Objective

Prolyl carboxypeptidase (PRCP) plays a role in the regulation of energy metabolism by inactivating hypothalamic α-melanocyte stimulating hormone (α-MSH) levels. Although detected in the arcuate nucleus, limited PRCP expression has been observed in the arcuate POMC neurons, and its site of action in regulating metabolism is still ill-defined.

Methods

We performed immunostaining to assess the localization of PRCP in arcuate Neuropeptide Y/Agouti-related Peptide (NPY/AgRP) neurons. Hypothalamic explants were then used to assess the intracellular localization of PRCP and its release at the synaptic levels. Finally, we generated a mouse model to assess the role of PRCP in NPY/AgRP neurons of the arcuate nucleus in the regulation of metabolism.

Results

Here we show that PRCP is expressed in NPY/AgRP-expressing neurons of the arcuate nucleus. In hypothalamic explants, stimulation by ghrelin increased PRCP concentration in the medium and decreased PRCP content in synaptic extract, suggesting that PRCP is released at the synaptic level. In support of this, hypothalamic explants from mice with selective deletion of PRCP in AgRP neurons (Prcp^AgRPKO) showed reduced ghrelin-induced PRCP concentration in the medium compared to controls mice. Furthermore, male Prcp^AgRPKO mice had decreased body weight and fat mass compared to controls. However, this phenotype was sex-specific as female Prcp^AgRPKO mice show metabolic differences only when challenged by high fat diet feeding. The improved metabolism of Prcp^AgRPKO mice was associated with reduced food intake and increased energy expenditure, locomotor activity, and hypothalamic α-MSH levels. Administration of SHU9119, a potent melanocortin receptor antagonist, selectively in the PVN of Prcp^AgRPKO male mice increased food intake to a level similar to that of control mice.

Conclusions

Altogether, our data indicate that PRCP is released at the synaptic levels and that PRCP in AgRP neurons contributes to the modulation of α-MSH degradation and related metabolic control in mice.