title: Airway smooth muscle dysfunction precedes teratogenic congenital diaphragmatic hernia and may contribute to hypoplastic lung morphogenesis.
abstract: Fetal intervention aims to improve lung growth and survival in congenital diaphragmatic hernia (CDH). Airway smooth muscle (ASM) is important in lung development: ASM progenitors produce a key growth factor for lung morphogenesis (fibroblast growth factor 10); ASM contractility is also coupled to growth. ASM hyperreactivity occurs in postnatal CDH and may exacerbate barotrauma via impaired lung compliance. We hypothesize that ASM hyperreactivity and its sequelae are based on an early developmental lesion of ASM activity in hypoplastic lung. Sprague-Dawley rats were fed 100 mg nitrofen on Day 9.5 of pregnancy to induce lung hypoplasia in offspring (controls had vehicle alone). Normal and hypoplastic lung primordia were cultured from Day 13.5 of gestation at 37 degrees C in 5% CO(2) and loaded at 54 or 78 h with Ca(2+)-sensitive indicators: Fluo-4 for confocal imaging and Indo-1 or Fura-2 for photometric measurements of [Ca(2+)](i). Hypoplastic lung features spontaneous propagating ASM Ca(2+) transients with reduced frequency, increased amplitude, and significantly prolonged plateau duration, relative to control lung. Nonetheless, hypoplastic lung exhibits normal requirement for extracellular calcium entry and intracellular calcium release in initiation and regulation of ASM Ca(2+) waves. Early ASM dysfunction in lung hypoplasia is apparent as specific anomalies of Ca(2+) transients that indicate a problem with plasmalemmal ion channels/action potential generation. Elucidation of such an ASM lesion may allow pharmacologic amelioration not only of ASM hyperreactivity and its sequelae, but also of hypoplastic lung growth itself.
authors.affiliation: Medical Research Council Clinical Training Fellow, Division of Child Health, School of Reproductive and Developmental Medicine, University of Liverpool, Liverpool L69 3BX, UK. N.C.Featherstone@Liverpool.ac.uk
authors.forename: Neil C
authors.initials: NC
authors.lastname: Featherstone
authors.forename: Marilyn G
authors.initials: MG
authors.lastname: Connell
authors.forename: David G
authors.initials: DG
authors.lastname: Fernig
authors.forename: Susan
authors.initials: S
authors.lastname: Wray
authors.forename: Theodor V
authors.initials: TV
authors.lastname: Burdyga
authors.forename: Paul D
authors.initials: PD
authors.lastname: Losty
authors.forename: Edwin C
authors.initials: EC
authors.lastname: Jesudason
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology; D002217:Carbachol / Q000378:metabolism; D018679:Cholinergic Agonists / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D005865:Gestational Age; D006548:Hernia, Diaphragmatic / Q000473:pathology / Q000503:physiopathology*; D006801:Humans; D008168:Lung / Q000033:anatomy & histology / Q000196:embryology / Q000254:growth & development / Q000473:pathology; D008170:Lung Compliance; D009024:Morphogenesis; D009130:Muscle, Smooth / Q000187:drug effects / Q000502:physiology*; D010647:Phenyl Ethers / Q000494:pharmacology; D011189:Potassium Chloride / Q000378:metabolism; D011247:Pregnancy; D011297:Prenatal Exposure Delayed Effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D015619:Respiratory System Abnormalities / Q000473:pathology / Q000503:physiopathology*; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D046509:Tissue Culture Techniques
chemical_list: D018679:Cholinergic Agonists; D005456:Fluorescent Dyes; D010647:Phenyl Ethers; D011189:Potassium Chloride; D002217:Carbachol; C007350:nitrofen; D002118:Calcium