Cystic Fibrosis Transmembrane Conductance Regulator Modulates Synaptic Chloride Homeostasis in Motoneurons of the Rat Spinal Cord During Neonatal Development

Cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-regulated Cl- channel functional in neonatal rat spinal motoneurons. The present study investigated the developmental (P1-P8) expression of CFTR, its impact on motoneuron excitability and Cl- homeostasis in relation to canonical Cl- transporters. The Cl- outward transporter KCC2 gene was upregulated in females over males and increased from P1 to P8. The gene activities of the Cl- inward transporter NKCC1 and CFTR were positively correlated and grew between P1 and P8. P1 motoneuronal somata were immunopositive for CFTR whose expression later (P8) extended to cell processes. KCC2 immunopositivity outlined somata and cell processes at P1 and P8. Electrophysiological recording with sharp electrodes showed that the CFTR blocker glibenclamide increased motoneuron input resistance, suggesting functional CFTR in P1-P8 motoneurons. Whole cell patchclamping of spinal motoneurons to study CFTR contribution to postnatal synaptic Cl- regulation indicated that glibenclamide or the selective CFTR blocker diphenylamine-2,20-dicarboxylic acid produced a negative shift in GABA/glycine reversal potential (EGABA/Gly) of spontaneously occurring synaptic events measured after block of excitatory transmission. A similar effect on EGABA/Gly was induced by the NKCC1 inhibitor bumetanide. A 3D reconstructed motoneuron model suggested that CFTR activity contributes to set the EGABA/Gly positive to the resting potential. The functional outcome of these Cl- mediated synaptic events depended not only on the postnatal age of the animal but also on their timing with respect to the excitatory synaptic signals. We propose that CFTR operated together with NKCC1 to produce depolarizing GABA/glycine mediated synaptic events. (C) 2010 Wiley Periodicals, Inc. Develop Neurobiol 71: 253-268, 2011