23 January 1997, Volume 120, Issue 3, Pages 481 – 487

Article

Characterization of Y3 receptor-mediated synaptic inhibition by chimeric neuropeptide Y-peptide YY peptides in the rat brainstem Steven R. Glaum1, Richard J. Miller2, Hyewhon Rhim2, Derek Maclean3,6, Lynn M. Georgic3, Robert G. MacKenzie3 & Lars Grundemar1,5 1Department of Physiology, Northwestern University Medical School, Chicago, IL 60611     2Department of Pharmacological and Physiological Sciences, University of Chicago, Chicago, IL 60637     3Parke Davies Pharmaceutical Research Division, Warner Lambert Company, Ann Arbour, MI 48105, U.S.A.     4Department of Clinical Pharmacology, Lund University Hospital, S-221 85 Lund, Sweden     5Author for correspondence 6Present address: Affymax Research Institute, Palo Alto, CA 95051, U.S.A.

Keywords

Neuropeptide Y;   peptide YY;   chimeric peptides;   neuropeptide Y receptors;   nucleus tractus solitarius;   postsynaptic currents;   electrophysiology;   ligand binding;   rat brainstem

Abstract

1   Neuropeptide Y (NPY) and peptide YY (PYY) act at receptors referred to as Y1 and Y2, while the Y3 receptor is specific to NPY and does not recognize PYY. The effects of NPY, its related peptides and a series of newly constructed chimeric NPY-PYY peptides were examined on excitatory and inhibitory postsynaptic currents (e.p.s.cs and i.p.s.cs, respectively) in rat dorsomedial nucleus tractus solitarius (NTS) neurones recorded in coronal brainstem slices. Monosynaptic activity was evoked by electrical stimulation in the region of the tractus solitarius. 2   NPY (5 – 500 nM) inhibited e.p.s.cs and i.p.s.cs in a concentration-dependent manner. In contrast, PYY (500 nM) failed to affect either e.p.s.cs or i.p.s.cs. The N- and C-terminal parts of a series of chimeric NPY-PYY peptides were joined at positions where NPY and PYY sequences differ. In binding experiments the chimeric peptides were all about equipotent with NPY and PYY in displacing [125I]-PYY from Y1 and Y2 binding sites on SK-N-MC cells and rat hippocampus respectively. 3   In the whole cell voltage clamp recordings of NTS neurones, NPY(1 – 23)-PYY(24 – 36) and NPY(1 – 14)-PYY(15 – 36) evoked a concentration-dependent inhibition of e.p.s.cs and i.p.s.cs, while NPY(1 – 7)-PYY(8 – 36) and NPY(1 – 3)-PYY(4 – 36) were inactive. The only differences in amino acid residues between NPY(1 – 14)-PYY(15 – 36) and NPY(1 – 7)-PYY(8 – 36) reside in positions 13 and 14. 4   Furthermore, [Pro34]NPY (500 nM) was equivalent in potency to NPY itself at inhibiting monosynaptic transmission in NTS, while [Leu31,Pro34]NPY and pancreatic polypeptide (both at 500 nM) failed to affect synaptic transmission. 5   The present study has shown that NPY acts at Y3 receptors to suppress both excitatory and inhibitory currents in the NTS. The different efficacy of the chimeric NPY-PYY peptides suggests that positions 13 and 14 are of great importance for Y3 receptor recognition. Finally, this receptor type readily recognizes [Pro34]NPY, but not [Leu31,Pro34]NPY.

Received 17 June, 96; Revised 23 September, 96; Accepted 1 October, 96

^© Macmillan Publishers Ltd 1997