Acute and chronic efficacy of bumetanide in an in vitro model of posttraumatic epileptogenesis

CNS Neurosci Ther. 2015 Feb;21(2):173-80. doi: 10.1111/cns.12369. Epub 2014 Dec 12.


Background: Seizures triggered by acute injuries to the developing brain respond poorly to first-line medications that target the inhibitory chloride-permeable GABAA receptor. Neuronal injury is associated with profound increases in cytoplasmic chloride ([Cl(-)]i) resulting in depolarizing GABA signaling, higher seizure propensity and limited efficacy of GABAergic anticonvulsants. The Na(+)-K(+)-2Cl(-) (NKCC1) cotransporter blocker bumetanide reduces [Cl(-)]i and causes more negative GABA equilibrium potential in injured neurons. We therefore tested both the acute and chronic efficacy of bumetanide on early posttraumatic ictal-like epileptiform discharges and epileptogenesis.

Methods: Acute hippocampal slices were used as a model of severe traumatic brain injury and posttraumatic epileptogenesis. Hippocampal slices were then incubated for 3 weeks. After a 1-week latent period, slice cultures developed chronic spontaneous ictal-like discharges. The anticonvulsant and anti-epileptogenic efficacy of bumetanide, phenobarbital, and the combination of these drugs was studied.

Results: Bumetanide reduced the frequency and power of early posttraumatic ictal-like discharges in vitro and enhanced the anticonvulsant efficacy of phenobarbital. Continuous 2-3 weeks administration of bumetanide as well as phenobarbital in combination with bumetanide failed to prevent posttraumatic ictal-like discharges and epileptogenesis.

Conclusions: Our data demonstrate a persistent contribution of NKCC1 cotransport in posttraumatic ictal-like activity, presumably as a consequence of chronic alterations in neuronal chloride homeostasis and GABA-mediated inhibition. New strategies for more effective reduction in posttraumatic and seizure-induced [Cl(-)]i accumulation could provide the basis for effective treatments for posttraumatic epileptogenesis and the resultant seizures.

Keywords: Bumetanide; NKCC1; Trauma; phenobarbital; posttraumatic seizures.

Publication types

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

MeSH terms

  • Action Potentials / drug effects
  • Action Potentials / genetics
  • Analysis of Variance
  • Animals
  • Anticonvulsants / pharmacology
  • Bumetanide / pharmacology*
  • Chlorides / metabolism
  • Dose-Response Relationship, Drug
  • Hippocampus / drug effects*
  • In Vitro Techniques
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Mutation / genetics
  • Organ Culture Techniques
  • Phenobarbital / pharmacology
  • Receptors, Immunologic / genetics
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sodium Potassium Chloride Symporter Inhibitors / pharmacology*
  • Time Factors


  • Anticonvulsants
  • CLM-1 protein, mouse
  • Chlorides
  • Receptors, Immunologic
  • Recombinant Fusion Proteins
  • Sodium Potassium Chloride Symporter Inhibitors
  • clomeleon
  • Bumetanide
  • Phenobarbital