Roles for Prlhr/GPR10 and Npffr2/GPR74 in Feeding Responses to PrRP.

Several groups of neurons in the NTS suppress food intake, including Prlh-expressing neurons (NTS cells). Not only does the artificial activation of NTS cells decrease feeding, but also the expression of Prlh (which encodes the neuropeptide PrRP) and neurotransmission by NTS neurons contributes to the restraint of food intake and body weight, especially in animals fed a high fat diet (HFD). We used animals lacking PrRP receptors GPR10 and/or GRP74 (encoded by Prlhr and Npffr2, respectively) to determine roles for each in the restraint of food intake and body weight by the increased expression of Prlh in NTS neurons (NTS mice) and in response to the anorectic PrRP analog, p52.

Streamlined single-molecule RNA-FISH of core clock mRNAs in clock neurons in whole mount brains.

Circadian clocks are ∼24-h timekeepers that control rhythms in almost all aspects of our behavior and physiology. While it is well known that subcellular localization of core clock proteins plays a critical role in circadian regulation, very little is known about the spatiotemporal organization of core clock mRNAs and its role in generating ∼24-h circadian rhythms. Here we describe a streamlined single molecule Fluorescence Hybridization (smFISH) protocol and a fully automated analysis pipeline to precisely quantify the number and subcellular location of mRNAs of , a core circadian transcription factor, in individual clock neurons in whole mount adult brains.