Inflammation Enhances Y1 Receptor Signaling, Neuropeptide Y-mediated Inhibition of Hyperalgesia, and Substance P Release From Primary Afferent Neurons

Neuroscience. 2014 Jan 3;256:178-94. doi: 10.1016/j.neuroscience.2013.10.054. Epub 2013 Oct 31.


Neuropeptide Y (NPY) is present in the superficial laminae of the dorsal horn and inhibits spinal nociceptive processing, but the mechanisms underlying its anti-hyperalgesic actions are unclear. We hypothesized that NPY acts at neuropeptide Y1 receptors in the dorsal horn to decrease nociception by inhibiting substance P (SP) release, and that these effects are enhanced by inflammation. To evaluate SP release, we used microdialysis and neurokinin 1 receptor (NK1R) internalization in rat. NPY decreased capsaicin-evoked SP-like immunoreactivity in the microdialysate of the dorsal horn. NPY also decreased non-noxious stimulus (paw brush)-evoked NK1R internalization (as well as mechanical hyperalgesia and mechanical and cold allodynia) after intraplantar injection of carrageenan. Similarly, in rat spinal cord slices with dorsal root attached, [Leu(31), Pro(34)]-NPY inhibited dorsal root stimulus-evoked NK1R internalization. In rat dorsal root ganglion neurons, Y1 receptors colocalized extensively with calcitonin gene-related peptide (CGRP). In dorsal horn neurons, Y1 receptors were extensively expressed and this may have masked the detection of terminal co-localization with CGRP or SP. To determine whether the pain inhibitory actions of Y1 receptors are enhanced by inflammation, we administered [Leu(31), Pro(34)]-NPY after intraplantar injection of complete Freund's adjuvant (CFA) in rat. We found that [Leu(31), Pro(34)]-NPY reduced paw clamp-induced NK1R internalization in CFA rats but not uninjured controls. To determine the contribution of increased Y1 receptor-G protein coupling, we measured [(35)S]GTPγS binding simulated by [Leu(31), Pro(34)]-NPY in mouse dorsal horn. CFA inflammation increased the affinity of Y1 receptor G-protein coupling. We conclude that Y1 receptors contribute to the anti-hyperalgesic effects of NPY by mediating the inhibition of SP release, and that Y1 receptor signaling in the dorsal horn is enhanced during inflammatory nociception.

Keywords: 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid; ANOVA; CFA; CGRP; DRG; G-protein; HEPES; IB4; NK1R; NPY; PBS; RIA; SP; SP-LI; SP-like immunoreactivity; aCSF; analysis of variance; artificial cerebrospinal fluid; calcitonin gene-related peptide; capsaicin; complete Freund’s adjuvant; dorsal root ganglion; isolectin B4; isolectin-B4; neurokinin 1 receptor; neurokinin-1 receptor; neuropeptide Y; pain; phosphate-buffered saline; radioimmunoassay; substance P.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Freund's Adjuvant / pharmacology
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacokinetics
  • Hyperalgesia / etiology*
  • Hyperalgesia / pathology
  • In Vitro Techniques
  • Inflammation / chemically induced
  • Inflammation / complications*
  • Inflammation / pathology
  • Male
  • Neurons, Afferent / drug effects
  • Neurons, Afferent / physiology*
  • Neuropeptide Y / metabolism*
  • Pain Measurement
  • Pain Threshold / drug effects
  • Protein Binding / drug effects
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Neurokinin-1 / metabolism
  • Receptors, Neuropeptide Y / metabolism*
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • Spinal Cord / pathology*
  • Spinal Nerve Roots / physiology
  • Substance P / metabolism*
  • Sulfur Isotopes / pharmacokinetics


  • Neuropeptide Y
  • Receptors, Neurokinin-1
  • Receptors, Neuropeptide Y
  • Sulfur Isotopes
  • Substance P
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Freund's Adjuvant