Retrograde Viral Vector-Mediated Inhibition of Pontospinal Noradrenergic Neurons Causes Hyperalgesia in Rats

J Neurosci. 2009 Oct 14;29(41):12855-64. doi: 10.1523/JNEUROSCI.1699-09.2009.

Abstract

Pontospinal noradrenergic neurons form a component of an endogenous analgesic system and represent a potential therapeutic target. We tested the principle that genetic manipulation of their excitability can alter nociception using an adenoviral vector (AVV-PRS-hKir(2.1)) containing a catecholaminergic-selective promoter (PRS) to retrogradely transduce and inhibit the noradrenergic neurons projecting to the lumbar dorsal horn through the expression of a potassium channel (hKir(2.1)). Expression of hKir(2.1) in catecholaminergic PC12 cells hyperpolarized the membrane potential and produced a barium-sensitive inward rectification. LC neurons transduced by AVV-PRS-hKir(2.1) in slice cultures also showed barium-sensitive inward rectification and reduced spontaneous firing rate (median 0.2 Hz; n = 19 vs control 1.0 Hz; n = 18, p < 0.05). Pontospinal noradrenergic neurons were retrogradely transduced in vivo by injection of AVV into the lumbar dorsal horn (L4-5). Rats transduced with AVV-PRS-hKir(2.1) showed thermal but not mechanical hyperalgesia. Similar selective augmentation of thermal hyperalgesia was seen in the CFA-inflammatory pain model after AVV-PRS-hKir(2.1). In the formalin test, rats transduced with hKir(2.1) showed enhanced nocifensive behaviors (both Phase I and II, p < 0.05, n = 11/group) and increased c-Fos-positive cells in the lumbar dorsal horn. Transduction with AVV-PRS-hKir(2.1) before spared nerve injury produced no change in tactile or cold allodynia. Thus, the selective genetic inhibition of approximately 150 pontospinal noradrenergic neurons produces a modality-specific thermal hyperalgesia, increased nocifensive behaviors, and spinal c-Fos expression in the formalin test, but not in the spared nerve injury model of neuropathic pain, indicating that these neurons exert a selective tonic restraining influence on in vivo nociception.

MeSH terms

  • Adenoviridae / genetics
  • Animals
  • Animals, Newborn
  • Catecholamines / genetics
  • Cell Count / methods
  • Disease Models, Animal
  • Dopamine beta-Hydroxylase / metabolism
  • Genetic Vectors / genetics*
  • Green Fluorescent Proteins / genetics
  • Hyperalgesia / etiology*
  • Hyperalgesia / pathology*
  • In Vitro Techniques
  • Laminectomy / methods
  • Male
  • Membrane Potentials / genetics
  • Membrane Potentials / physiology
  • Motor Activity / genetics
  • Neurons / metabolism*
  • Norepinephrine / metabolism*
  • PC12 Cells
  • Pain Measurement / methods
  • Pain Threshold / physiology
  • Patch-Clamp Techniques / methods
  • Peripheral Nervous System Diseases / physiopathology
  • Pons / pathology*
  • Potassium Channels, Inwardly Rectifying / genetics
  • Promoter Regions, Genetic / genetics
  • Proto-Oncogene Proteins c-fos / metabolism
  • Rats
  • Rats, Wistar
  • Transfection / methods

Substances

  • Catecholamines
  • Kir2.1 channel
  • Potassium Channels, Inwardly Rectifying
  • Proto-Oncogene Proteins c-fos
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Dopamine beta-Hydroxylase
  • Norepinephrine