title: Inhibition of carbonic anhydrase augments GABAA receptor-mediated analgesia via a spinal mechanism of action.
abstract: Peripheral nerve injury (PNI) negatively influences spinal gamma-aminobutyric acid (GABA)ergic networks via a reduction in the neuron-specific potassium-chloride (K(+)-Cl(-)) cotransporter (KCC2). This process has been linked to the emergence of neuropathic allodynia. In vivo pharmacologic and modeling studies show that a loss of KCC2 function results in a decrease in the efficacy of GABAA-mediated spinal inhibition. One potential strategy to mitigate this effect entails inhibition of carbonic anhydrase activity to reduce HCO3(-)-dependent depolarization via GABAA receptors when KCC2 function is compromised. We have tested this hypothesis here. Our results show that, similarly to when KCC2 is pharmacologically blocked, PNI causes a loss of analgesic effect for neurosteroid GABAA allosteric modulators at maximally effective doses in naïve mice in the tail-flick test. Remarkably, inhibition of carbonic anhydrase activity with intrathecal acetazolamide rapidly restores an analgesic effect for these compounds, suggesting an important role of carbonic anhydrase activity in regulating GABAA-mediated analgesia after PNI. Moreover, spinal acetazolamide administration leads to a profound reduction in the mouse formalin pain test, indicating that spinal carbonic anhydrase inhibition produces analgesia when primary afferent activity is driven by chemical mediators. Finally, we demonstrate that systemic administration of acetazolamide to rats with PNI produces an antiallodynic effect by itself and an enhancement of the peak analgesic effect with a change in the shape of the dose-response curve of the α1-sparing benzodiazepine L-838,417. Thus, carbonic anhydrase inhibition mitigates the negative effects of loss of KCC2 function after nerve injury in multiple species and through multiple administration routes resulting in an enhancement of analgesic effects for several GABAA allosteric modulators. We suggest that carbonic anhydrase inhibitors, many of which are clinically available, might be advantageously employed for the treatment of pathologic pain states.
  
  
  CONCLUSIONS
  Using behavioral pharmacology techniques, we show that spinal GABAA-mediated analgesia can be augmented, especially following nerve injury, via inhibition of carbonic anhydrases. Carbonic anhydrase inhibition alone also produces analgesia, suggesting these enzymes might be targeted for the treatment of pain.
authors.affiliation: Department of Pharmacology, The University of Arizona School of Medicine, Tucson, Arizona.
authors.forename: Marina N
authors.initials: MN
authors.lastname: Asiedu
authors.affiliation: Department of Pharmacology, The University of Arizona School of Medicine, Tucson, Arizona.
authors.forename: Galo L
authors.initials: GL
authors.lastname: Mejia
authors.affiliation: Friedrich-Schiller-University Jena, Institute of Human Genetics, Jena University Hospital, Jena, Germany.
authors.forename: Christian A
authors.initials: CA
authors.lastname: Hübner
authors.affiliation: Department of Biosciences and Neuroscience Center, University of Helsinki, Helsinki, Finland.
authors.forename: Kai
authors.initials: K
authors.lastname: Kaila
authors.affiliation: Department of Pharmacology, The University of Arizona School of Medicine, Tucson, Arizona; Graduate Interdisciplinary Program in Neuroscience, The University of Arizona School of Medicine, Tucson, Arizona. Electronic address: tjprice@email.arizona.edu.
authors.forename: Theodore J
authors.initials: TJ
authors.lastname: Price
mesh_terms: D000086:Acetazolamide / Q000494:pharmacology; D000818:Animals; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology*; D002256:Carbonic Anhydrases / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009437:Neuralgia / Q000378:metabolism*; D059348:Peripheral Nerve Injuries / Q000150:complications / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000378:metabolism*; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism*
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D011963:Receptors, GABA-A; D002256:Carbonic Anhydrases; D000086:Acetazolamide
keywords: Gamma-aminobutyric acid; carbonic anhydrase; neuron-specific K(+)-Cl(–) cotransporter; neuropathic pain; spinal analgesia