Expression of CFTR and Cl<sup>−</sup>conductances in cells of pulmonary neuroepithelial bodies

Yeger, Herman; Pan, Jie; Fu, Xiao Wen; Bear, Christine E.; Cutz, Ernest

doi:10.1152/ajplung.2001.281.3.l713

Cited by 20 publications

(14 citation statements)

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“…To enrich the mixed airway cell preparation in PNEC/NEB, we subjected the cell preparation to three courses of differential plating in serum-free medium. The carcinoid tumor cell line H727 (related to classic small cell lung cancer) was obtained from American Type Culture Collection and cultured as previously reported (44). Both cell preparations (enriched rabbit fetal PNEC/NEB and H727 cells) were seeded onto collagen I BioFlex plates (Flexcell International, Hillsborough, NC) and maintained in RPMI 1640 plus 10% FBS and ␣-MEM medium with 10% FBS, respectively, at 37°C in a 5% CO 2 incubator.…”

Section: Methodsmentioning

confidence: 99%

“…To determine the effects of mechanical stretch on 5-HT synthesis in PNEC/NEB, we assessed the expression of TPH, the ratelimiting enzyme in 5-HT synthesis. The expression of TPH mRNA has been previously documented in both rabbit lung PNEC and PNEC-related tumor cell lines (13,31,44). We assessed expression of TPH at a protein level by performing Western blot analysis of cultures of PNEC from E26 rabbit fetal lung subjected to either cyclic mechanical stretch or hypoxia (Fig.…”

Section: Effect Of Mechanical Stimulation On 5-ht Releasementioning

confidence: 99%

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Mechanical stretch-induced serotonin release from pulmonary neuroendocrine cells: implications for lung development

Pan

Copland²,

Post³

et al. 2006

American Journal of Physiology-Lung Cellular and Molecular Phys

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(serotonin, and peptides (e.g., bombesin, calcitonin) with growth factor-like properties and are thought to play an important role in lung development. Because physical forces are essential for lung growth and development, we investigated the effects of mechanical strain on 5-HT release in PNEC freshly isolated from rabbit fetal lung and in the PNEC-related tumor H727 cell line. Cultures exposed to sinusoidal cyclic stretch showed a significant 5-HT release inhibitable with gadolinium chloride (10 nM), a blocker of mechanosensitive channels. In contrast to hypoxia (PO 2 ϳ 20 mmHg), stretch-induced 5-HT release was not affected by Ca 2ϩ -free medium or nifedipine (50 M), excluding the exocytic pathway. In H727 cells, stretch failed to release calcitonin, a peptide stored within dense core vesicles (DCV), whereas hypoxia caused massive calcitonin release. 5-HT released by mechanical stretch is derived predominantly from the cytoplasmic pool, because it is rapid (ϳ5 min) and is releasable from early (20 days of gestation) fetal PNEC containing few DCV. Both mechanical stretch and hypoxia upregulated expression of tryptophan hydroxylase, the rate-limiting enzyme of 5-HT synthesis. We conclude that mechanical strain is an important physiological stimulus for the release of 5-HT from PNEC via mechanosensitive channels with potential effects on lung development and resorption of lung fluid at the time of birth. mechanosensitive ion channels; amine storage pools; neuroendocrine cell secretion; hypoxia-induced secretion; fetal lung fluid THE SYSTEM OF PULMONARY NEUROENDOCRINE CELLS (PNEC) is composed of amine (serotonin, 5-HT)-and peptide (e.g., bombesin, calcitonin)-producing cells widely distributed within the airway mucosa of human and animal lungs (13,34,40). PNEC occur as single cells and as distinctive innervated clusters, neuroepithelial bodies (NEB). Whereas solitary PNEC populate the mucosa of the trachea and bronchi up to the terminal bronchioles, NEB are found only within intrapulmonary airways, where they appear concentrated at airway branch points (7). Pulmonary NEB function as airway O 2 sensors, because they express a membrane-bound O 2 -sensing molecular complex composed of an O 2 -sensitive K ϩ channel coupled to an O 2 -sensing protein, NADPH oxidase (9, 10, 45). Hypoxia activates the O 2 sensor, leading to Ca 2ϩ -dependent exocytosis of dense core vesicles (DCV), the storage site of amine and peptides, and the release of these mediators acting locally or via vagal afferents (9, 10). Hypoxia induced 5-HT release from NEB cells is dose dependent and occurs within the physiological range expected in the airway (PO 2 ϳ 95 mmHg) (17). The precise role of solitary PNEC and whether they are functionally distinct from NEB is not known, although they share identical neuroendocrine and molecular markers as well as innervation (13,34,40). PNEC/NEB appear prominent in fetal and neonatal lungs but are less conspicuous in the lungs of adults because of a "dilutional" effect of postnatal lung growth (20).PNEC are th...

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Section: Methodsmentioning

confidence: 99%

Section: Effect Of Mechanical Stimulation On 5-ht Releasementioning

confidence: 99%

Mechanical stretch-induced serotonin release from pulmonary neuroendocrine cells: implications for lung development

Pan

Copland²,

Post³

et al. 2006

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Urine GRP levels are high postnatally in children with CF, in contrast to the decline in normal infants (76). PNECs express CFTR at the apical membrane, suggesting that NEBs could contribute to CF lung disease, including the early stages before establishment of chronic infection and progressive lung disease (128, 129). Although PNECs, airway innervation, and smooth muscle are altered in Cftr-null mice (130), it remains possible that PNEC abnormalities are secondary to infection and/or inflammation.…”

Section: Potential Relevance Of Grp To Other Pediatric Lung Diseasesmentioning

confidence: 99%

Oxygen, Gastrin-Releasing Peptide, and Pediatric Lung Disease: Life in the Balance

Sunday

2014

Front. Pediatr.

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Excessive oxygen (O2) can cause tissue injury, scarring, aging, and even death. Our laboratory is studying O2-sensing pulmonary neuroendocrine cells (PNECs) and the PNEC-derived product gastrin-releasing peptide (GRP). Reactive oxygen species (ROS) generated from exposure to hyperoxia, ozone, or ionizing radiation (RT) can induce PNEC degranulation and GRP secretion. PNEC degranulation is also induced by hypoxia, and effects of hypoxia are mediated by free radicals. We have determined that excessive GRP leads to lung injury with acute and chronic inflammation, leading to pulmonary fibrosis (PF), triggered via ROS exposure or by directly treating mice with exogenous GRP. In animal models, GRP-blockade abrogates lung injury, inflammation, and fibrosis. The optimal time frame for GRP-blockade and the key target cell types remain to be determined. The concept of GRP as a mediator of ROS-induced tissue damage represents a paradigm shift about how O2 can cause injury, inflammation, and fibrosis. The host PNEC response in vivo may depend on individual ROS sensing mechanisms and subsequent GRP secretion. Ongoing scientific and clinical investigations promise to further clarify the molecular pathways and clinical relevance of GRP in the pathogenesis of diverse pediatric lung diseases.

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Section: Cell Culturementioning

confidence: 99%

“…All other antibodies were purchased from commercial sources and applied as previously described (Yeger et al 2001). Immunocytochemical labeling was enhanced with the catalyzed reporter deposition (CARD) amplification method as previously described (Yeger et al 2001). Briefly, hydroxysuccinimide ester of rhodamine (TRITC-NHS) and biotin (Biotin-NHS) were coupled to tyramine according to Hopman et al (1998) to produce TRITC-tyramine and biotin-tyramine substrates for peroxidase reaction, The conjugate was utilized in the CARD system to amplify FORSE-1 immunofluorescent and immunohistochemical labeling, respectively.…”

Section: Cell Culturementioning

confidence: 99%

Neuronal Developmental Marker FORSE-1 Identifies a Putative Progenitor of the Pulmonary Neuroendocrine Cell Lineage During Lung Development

Pan

Yeger

Cutz

2002

J Histochem Cytochem.

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S U M M A R YThe FORSE-1 ( fore brain-s urface-e mbryonic) monoclonal antibody (MAb) recognizes a carbohydrate cell surface epitope related to the Lewis-X (LeX) and stage-specific embryonic antigens (SSEAs). In the developing CNS, the FORSE-1 epitope is believed to serve as a marker of progenitor cells. We studied the expression of the FORSE-1 epitope in pulmonary neuroendocrine cells (PNECs) and related neuroepithelial bodies (NEBs), cell types implicated in paracrine regulation of lung development. We used dual immunolabeling to identify PNECs/NEBs in tissue sections from developing rabbit fetal lungs and corresponding primary lung cell cultures. During the early stage (E16), the FORSE-1 MAb labeled primitive airway epithelium, whereas serotonin (5HT) immunoreactivity, a marker of PNEC/ NEB differentiation, was negative. After E18, FORSE-1 labeling became restricted to PNECs and NEBs, identified by co-expression with 5HT, then decreased coincident with an increase in 5HT. Expression of the FORSE-1 epitope correlated inversely with 5HT expression in PNEC/NEB cells. FORSE-1 immunoreactivity correlated with cell proliferation assessed by BrdU labeling. Downregulation of the FORSE-1 epitope correlated with maturation of PNECs/NEBs. The presence of few FORSE-1/5HT-positive cells in postnatal lung suggests retention of progenitors. The FORSE-1 epitope was associated with a high molecular weight (286 kD) glycoprotein that decreased with increasing gestational age, as demonstrated by immunoblotting. Overall expression of SSEA-1, -3, and -4 antigens was similar to FORSE-1/ 5HT, although the former was preferentially localized to neurite-like processes. Because the role of the FORSE-1 epitope in the CNS probably involves cell adhesion and differentiation, we propose a similar function in developing lung. The demonstration of LeX/SSEA antigen expression in the PNEC/NEB cell lineage underscores the importance of these cells in developing lung. Furthermore, the FORSE-1 antigen may identify committed progenitors of the PNEC/NEB cell system.

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Expression of CFTR and Cl⁻conductances in cells of pulmonary neuroepithelial bodies

Cited by 20 publications

References 74 publications

Mechanical stretch-induced serotonin release from pulmonary neuroendocrine cells: implications for lung development

Mechanical stretch-induced serotonin release from pulmonary neuroendocrine cells: implications for lung development

Oxygen, Gastrin-Releasing Peptide, and Pediatric Lung Disease: Life in the Balance

Neuronal Developmental Marker FORSE-1 Identifies a Putative Progenitor of the Pulmonary Neuroendocrine Cell Lineage During Lung Development

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Expression of CFTR and Cl−conductances in cells of pulmonary neuroepithelial bodies

Cited by 20 publications

References 74 publications

Mechanical stretch-induced serotonin release from pulmonary neuroendocrine cells: implications for lung development

Mechanical stretch-induced serotonin release from pulmonary neuroendocrine cells: implications for lung development

Oxygen, Gastrin-Releasing Peptide, and Pediatric Lung Disease: Life in the Balance

Neuronal Developmental Marker FORSE-1 Identifies a Putative Progenitor of the Pulmonary Neuroendocrine Cell Lineage During Lung Development

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Expression of CFTR and Cl⁻conductances in cells of pulmonary neuroepithelial bodies