CFTR Modulates Hypothalamic Neuron Excitability to Maintain Female Cycle

Cystic fibrosis transmembrane conductance regulator (CFTR), known as an epithelial Cl⁻ channel, is increasingly noted to be expressed in the nervous system, although whether and how it plays a role in neuronal excitability is unclear. Given the association of CFTR with fertility, we tested here possible involvement of CFTR in regulating hypothalamic neuron excitability. Patch-clamp and Ca²⁺ imaging showed that pharmacological inhibition of CFTR evoked electrical pulses and Ca²⁺ spikes in primary rat hypothalamic neurons, which was dependent on extracellular Cl⁻. Hypothalamic neurons in brain-slice preparations from adult female mice with CFTR mutation (DF508) exhibited significantly reduced electrical pulses as compared to the wild-type controls. Removal of extracellular Cl⁻ eliminated hypothalamic electrical pulses in the wild-type brain slices, which was reversible by subsequent addition of Cl⁻. In adult female mice, Ca²⁺ indicator (GCaMP6s)-based fiber-photometry showed that hypothalamic Ca²⁺ activities in vivo were enhanced at the proestrus/estrus phase as compared to the diestrus phase of the female cycle. Such estrus-associated hypothalamic activities were largely diminished in DF508 female mice, together with delayed puberty and disturbed female cycles. Therefore, these findings suggest a critical role of CFTR in modulating hypothalamic neuron excitability, which may account for the disturbed female cycles and reduced female fertility associated with CFTR mutations.