CFTR is functionally active in GnRH-expressing GT1–7 hypothalamic neurons

• DOI: 10.1152/ajpcell.1999.277.3.c563
• Publication Date: 2017-12-24T22:10:19Z
• Authors: Richard T. Weyler; Karin A. Yurko-Mauro; Ronald Rubenstein; Wouter J. W. Kollen; William Reenstra; Steven M. Altschuler; Marie Egan; Andrew E. Mulberg
• Keywords: Neurons; 0301 basic medicine; Reverse Transcriptase Polymerase Chain Reaction; Blotting, Western; Hypothalamus; Cystic Fibrosis Transmembrane Conductance Regulator; 3T3 Cells; Oligonucleotides, Antisense; Cell Line; Gonadotropin-Releasing Hormone; Mice; 03 medical and health sciences; Cyclic AMP; Animals; RNA, Messenger; In Situ Hybridization

We have demonstrated the expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, mRNA, and protein within the rat and human brains, in areas regulating sexual differentiation and function. We have found that GT1–7, a gonadotropin-releasing hormone (GnRH)-secreting hypothalamic neuronal cell line, expresses the CFTR gene, mRNA, and protein and cAMP-dependent 36Cl efflux. A linear 7-pS Cl− conductance, which is stimulated by ATP and cAMP analogs and inhibited by glibenclamide, consistent with CFTR activity, has been identified in GT1–7 cells. Antisense oligo(dN) generated against exon 10 of the CFTR gene transcript (mRNA) inhibit GnRH secretion into media [312 ± 73, 850 ± 150, 963 ± 304, and 912 ± 74 pg GnRH/4 × 106 cells for antisense, sense, missense, and no oligo(dN), respectively; P < 0.029 for antisense oligo(dN)-treated vs. normal cells]. No changes in intracellular synthesis of GnRH were noted [1,400 ± 371 and 1,395 ± 384 pg GnRH/4 × 106 cells for antisense and sense oligo(dN), respectively]. Antisense oligo(dN), but not sense or missense oligo(dN), inhibited cAMP-dependent36Cl efflux. The expression of CFTR protein, detected by Western blotting, was also inhibited 68% by preincubation of cells with antisense oligo(dN). GT1–7 hypothalamic neurons express the CFTR gene, mRNA, and protein, which modulate neurosecretion. Abnormal neuropeptide vesicle trafficking by mutant CFTR may help to explain some of the diverse manifestations of cystic fibrosis.