title: Stimulation of β2-adrenergic receptor increases CFTR function and decreases ATP levels in murine hematopoietic stem/progenitor cells.
abstract: BACKGROUND
  The chloride channel CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) is expressed by many cell types, including hematopoietic stem/progenitor cells (HSPCs). In this study, we sought to better comprehend the regulation of CFTR activity in HSPCs, namely by beta-adrenergic stimuli.
  
  
  METHODS
  The expression of β2-adrenergic receptor (β2-AR) in murine Sca-1(+) HSPCs was investigated by immunofluorescence/confocal microscopy and flow-cytometric analysis. Association with CFTR was assessed by immunoprecipitation. HSPCs were evaluated for ATP content and CFTR activity by means of luminometric and spectrofluorometric methods, respectively, upon stimulation with salbutamol.
  
  
  RESULTS
  HSPCs express β2-AR over the whole plasma membrane and are associated in cellula with both the immature and mature forms of CFTR. β2-AR was predominantly expressed by HSPCs with bigger size. CFTR channel activity was increased by salbutamol treatment and this activation was inhibited by either a specific CFTR inhibitor (CFTRinh172) or a β2-AR receptor inhibitor (ICI 118,551). Intracellular ATP levels were reduced by salbutamol stimulation and this effect was reversed when ICI 118,551 or CFTR inhibitors were present. A trend in the increase of extracellular ATP upon salbutamol stimulation was observed.
  
  
  CONCLUSIONS
  In HSPCs, CFTR is regulated by β2-adrenergic receptor stimulation determining intracellular ATP depletion.
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Teresa
authors.initials: T
authors.lastname: Trotta
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Guerra
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Donatella
authors.initials: D
authors.lastname: Piro
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: d'Apolito
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Piccoli
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Porro
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Ida
authors.initials: I
authors.lastname: Giardino
authors.affiliation: Laboratory of Preclinical and Translational Research, IRCCS-CROB Rionero in Vulture, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Lepore
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Castellani
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Sante
authors.initials: S
authors.lastname: Di Gioia
authors.affiliation: Istituto Zooprofilattico di Puglia e Basilicata, Foggia, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Petrella
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Angela Bruna
authors.initials: AB
authors.lastname: Maffione
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Casavola
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Nazzareno
authors.initials: N
authors.lastname: Capitanio
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy. Electronic address: massimo.conese@unifg.it.
authors.forename: Massimo
authors.initials: M
authors.lastname: Conese
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000420:Albuterol / Q000494:pharmacology*; D000704:Analysis of Variance; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002478:Cells, Cultured; D018118:Chloride Channels / Q000378:metabolism; D003550:Cystic Fibrosis / Q000378:metabolism / Q000503:physiopathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000187:drug effects* / Q000378:metabolism*; D004195:Disease Models, Animal; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal / Q000379:methods; D011897:Random Allocation; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013234:Stem Cells
chemical_list: D015415:Biomarkers; D018118:Chloride Channels; D018343:Receptors, Adrenergic, beta-2; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000255:Adenosine Triphosphate; D000420:Albuterol
keywords: ATP; Adrenergic receptor; CFTR; Hematopoietic stem/progenitor cells