title: Tumor Necrosis Factor-α Impairs Kisspeptin Signaling in Human Gonadotropin-Releasing Hormone Primary Neurons.
abstract: Inflammatory pathways may impair central regulatory networks involving gonadotropin-releasing hormone (GnRH) neuron activity. Studies in humans are limited by the lack of human GnRH neuron cell lines.
  
  
  
  To establish an in vitro model of human GnRH neurons and analyze the effects of proinflammatory cytokines.
  
  
  
  The primary human fetal hypothalamic cells (hfHypo) were isolated from 12-week-old fetuses. Responsiveness to kisspeptin, the main GnRH neurons' physiological regulator, was evaluated for biological characterization. Tumor necrosis factor alpha (TNF-α) was used as a proinflammatory stimulus. Main Outcome Measures: Expression of specific GnRH neuron markers by quantitative reverse transcription-polymerase chain reaction, flow cytometry, and immunocytochemistry analyses; and GnRH-releasing ability and electrophysiological changes in response to kisspeptin.
  
  
  
  The primary hfHypo showed a high percentage of GnRH-positive cells (80%), expressing a functional kisspeptin receptor (KISS1R) and able to release GnRH in response to kisspeptin. TNF-α exposure determined a specific inflammatory intracellular signaling and reduced GnRH secretion, KISS1R expression, and kisspeptin-induced depolarizing effect. Moreover, hfHypo possessed a primary cilium, whose assembly was inhibited by TNF-α.
  
  
  
  The hfHypo cells represent a novel tool for investigations on human GnRH neuron biology. TNF-α directly affects GnRH neuron function by interfering with KISS1R expression and ciliogenesis, thereby impairing kisspeptin signaling.
authors.affiliation: Section of Human Anatomy and Histology.
authors.forename: Erica
authors.initials: E
authors.lastname: Sarchielli
authors.affiliation: Sexual Medicine and Andrology Unit and.
authors.forename: Paolo
authors.initials: P
authors.lastname: Comeglio
authors.affiliation: Section of Physiological Sciences, and.
authors.forename: Roberta
authors.initials: R
authors.lastname: Squecco
authors.affiliation: Cell Therapy and Transfusion Medicine Unit, Department of Experimental and Clinical Medicine, University of Florence, 50134 Florence, Italy.
authors.forename: Lara
authors.initials: L
authors.lastname: Ballerini
authors.affiliation: Gastroenterology Unit, Department of Experimental and Clinical Biomedical Sciences "Mario Serio," University of Florence, 50134 Florence, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Mello
authors.affiliation: Section of Human Anatomy and Histology.
authors.forename: Giulia
authors.initials: G
authors.lastname: Guarnieri
authors.affiliation: Section of Physiological Sciences, and.
authors.forename: Eglantina
authors.initials: E
authors.lastname: Idrizaj
authors.affiliation: Cell Therapy and Transfusion Medicine Unit, Department of Experimental and Clinical Medicine, University of Florence, 50134 Florence, Italy.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Mazzanti
authors.affiliation: Sexual Medicine and Andrology Unit and.
authors.forename: Linda
authors.initials: L
authors.lastname: Vignozzi
authors.affiliation: Neurosurgery School of Tuscany, Department of Surgery and Translational Medicine, University of Florence, 50139 Florence, Italy; and.
authors.forename: Pasquale
authors.initials: P
authors.lastname: Gallina
authors.affiliation: Sexual Medicine and Andrology Unit and.|Istituto Nazionale Biostrutture e Biosistemi, 00136 Rome, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Maggi
authors.affiliation: Section of Human Anatomy and Histology.
authors.forename: Gabriella B
authors.initials: GB
authors.lastname: Vannelli
authors.affiliation: Section of Human Anatomy and Histology.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Morelli
mesh_terms: D002478:Cells, Cultured; D005260:Female; D005333:Fetus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D006801:Humans; D059648:Kisspeptins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D000074502:Receptors, Kisspeptin-1; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: C118688:KISS1R protein, human; D059648:Kisspeptins; D043562:Receptors, G-Protein-Coupled; D000074502:Receptors, Kisspeptin-1; D014409:Tumor Necrosis Factor-alpha; D007987:Gonadotropin-Releasing Hormone