title: Amylin receptor components and the leptin receptor are co-expressed in single rat area postrema neurons.
abstract: Amylin is a pancreatic β-cell hormone that acts as a satiating signal to inhibit food intake by binding to amylin receptors (AMYs) and activating a specific neuronal population in the area postrema (AP). AMYs are heterodimers that include a calcitonin receptor (CTR) subunit [CTR isoform a or b (CTRa or CTRb)] and a member of the receptor activity-modifying proteins (RAMPs). Here, we used single-cell quantitative polymerase chain reaction to assess co-expression of AMY subunits in AP neurons from rats that were injected with amylin or vehicle. Because amylin interacts synergistically with the adipokine leptin to reduce body weight, we also assessed the co-expression of AMY and the leptin receptor isoform b (LepRb) in amylin-activated AP neurons. Single cells were collected from Wistar rats and from transgenic Fos-GFP rats that express green fluorescent protein (GFP) under the control of the Fos promoter. We found that the mRNAs of CTRa, RAMP1, RAMP2 and RAMP3 were all co-expressed in single AP neurons. Moreover, most of the CTRa+ cells co-expressed more than one of the RAMPs. Amylin down-regulated RAMP1 and RAMP3 but not CTR mRNAs in AMY+ neurons, suggesting a possible negative feedback mechanism of amylin at its own primary receptors. Interestingly, amylin up-regulated RAMP2 mRNA. We also found that a high percentage of single cells that co-expressed all components of a functional AMY expressed LepRb mRNA. Thus, single AP cells expressed both AMY and LepRb, which formed a population of first-order neurons that presumably can be directly activated by amylin and, at least in part, also by leptin.
authors.affiliation: Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zürich (UZH), Winterthurerstrasse 260, CH-8057, Zürich, Switzerland.|Zürich Centre for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland.|Zürich Centre for Clinical Studies, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland.
authors.forename: Claudia G
authors.initials: CG
authors.lastname: Liberini
authors.affiliation: Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zürich (UZH), Winterthurerstrasse 260, CH-8057, Zürich, Switzerland.
authors.forename: Christina N
authors.initials: CN
authors.lastname: Boyle
authors.affiliation: School of Pharmacy, Pharmacology Unit, University of Camerino, Camerino, Italy.|Intramural Research Program, National Institutes of Health/National Institute on Drug Abuse, Baltimore, MD, USA.
authors.forename: Carlo
authors.initials: C
authors.lastname: Cifani
authors.affiliation: Intramural Research Program, National Institutes of Health/National Institute on Drug Abuse, Baltimore, MD, USA.
authors.forename: Marco
authors.initials: M
authors.lastname: Venniro
authors.affiliation: Intramural Research Program, National Institutes of Health/National Institute on Drug Abuse, Baltimore, MD, USA.
authors.forename: Bruce T
authors.initials: BT
authors.lastname: Hope
authors.affiliation: Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zürich (UZH), Winterthurerstrasse 260, CH-8057, Zürich, Switzerland.|Zürich Centre for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Lutz
mesh_terms: D000818:Animals; D031608:Area Postrema / Q000166:cytology / Q000378:metabolism*; D025461:Feedback, Physiological; D005260:Female; D058228:Islet Amyloid Polypeptide / Q000494:pharmacology; D020738:Leptin / Q000494:pharmacology; D008297:Male; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D058287:Receptors, Islet Amyloid Polypeptide / Q000235:genetics / Q000378:metabolism*; D054411:Receptors, Leptin / Q000235:genetics / Q000378:metabolism*
chemical_list: D058228:Islet Amyloid Polypeptide; D020738:Leptin; D021122:Protein Subunits; D012333:RNA, Messenger; D058287:Receptors, Islet Amyloid Polypeptide; D054411:Receptors, Leptin
keywords: calcitonin receptor; laser capture microdissection; receptor activity-modifying protein 1; receptor activity-modifying protein 2; receptor activity-modifying protein 3