Innervation of Pulmonary Neuroendocrine Cells and Neuroepithelial Bodies in Developing Rabbit Lung

Pan, Jie; Yeger, Herman; Cutz, Ernest

doi:10.1177/002215540405200309

Cited by 60 publications

(44 citation statements)

References 32 publications

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“…Such an arrangement would favor transmission of physical forces generated by the breathing movements. Similar cells also are found in the epithelium of larger airways in rabbit fetal/neonatal lungs with most of them innervated by a complex network of interconnecting fine varicose nerve fibers originating in the submucosa (data not shown) (32). PNEC/ NEB isolated from early (E20) or late (E 26) rabbit fetal lungs and grown on collagen I BioFlex membranes appeared well preserved and were similar to other in vitro preparations used in our previous studies (45).…”

Section: Immunolocalization Of 5-htsupporting

confidence: 76%

“…Such functions may be developmentally regulated, because in PNEC from early stages of lung development, the cytoplasmic pool of 5-HT predominates. The role of locally released 5-HT in early stages of lung development may involve effects on cell proliferation and differentiation as well as that of a neurotransmitter signaling via emerging PNEC innervation (32). During the later stages of lung development, the stretch-induced 5-HT release could be complemented with hypoxia-stimulated exocytosis mechanism once the necessary cell machinery develops for the synthesis and storage of amine and peptides within DCV.…”

Section: Discussionmentioning

confidence: 99%

“…The following primary antibodies were used: monoclonal antibody against serotonin (5-HT) (7) and calcitonin (11,12,39). The immunohistochemistry procedures using the avidin-biotin complex method (Vectastain ABC kit; Vector Laboratories, Burlingame, CA) or the indirect immunofluorescence method were carried out as previously reported (7,11,12,32). As negative control, primary antibody was omitted.…”

Section: Methodsmentioning

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: Immunolocalization Of 5-htsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

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

Self Cite

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“…PNECs/NEBs can release many bioactive substances with growth factors and mitogenic properties, such as bombesin/ Gastrin-Releasing Peptide (GRP), Calcitonin Gene-Related Peptide (CGRP), and 5-hydroxytryptamine (5-HT) [20]. These substances play an important role in lung morphogenesis, which is why NEBs prevail in fetal and neonatal lungs [21]. Morphogenesis is a process much like tissue remodeling, which is an important step in PF.…”

Section: The Nebsmentioning

confidence: 99%

Neuro-immune interactions during development of pulmonary fibrosis

Jain¹,

Yu²

2017

Pulm Crit Care Med

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Pulmonary neuroendocrine cells are widely distributed throughout the airway mucosa of mammals. They may exist as solitary cells or aggregate to form NeuroEpithelial Bodies (NEBs), which are important in early lung development. Pulmonary Fibrosis (PF) is a devastating disease caused by various pulmonary insults. The lung repair after injury is a complex process which involves a network of interactions among mediators, cytokines, and growth factors of inflammatory, endothelial, and epithelial cell origin. These interactions lead to lung remodeling, deposition of collagen, limitation of gas exchange, and dyspnea. The vagus nerve is related to each component of this network. Increasing evidence indicates that the NEB-Vagal System (NVS) plays an important role in neuro-immune interactions. NEBs are richly innervated and connect with the central nervous system through vagal afferent and efferent nerves. Stimulation of the vagus nerve in patients significantly increases the levels of TGF-β in plasma, a substance that promotes fibrosis. Based on current information, lung injury may activate the NVS, which then leads to PF via fibrogenic cells and molecules. Therefore, over-activation of the NVS could contribute to disease progression.

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“…13 The variable excitation wavelengths of the krypton/argon laser were 488 nm for FITC conjugate, 568 nm for Texas Red complex and 695 nm for Alexa Fluor 680 conjugate/RedDot-2 (nuclear counterstaining).…”

Section: Confocal Microscopymentioning

confidence: 99%

Multilabel immunofluorescence and antigen reprobing on formalin-fixed paraffin-embedded sections: novel applications for precision pathology diagnosis

et al. 2016

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We report new methods for multilabel immunofluorescence (MIF) and reprobing of antigen epitopes on the same formalin-fixed paraffin-embedded (FFPE) sections. The MIF method includes an antigen-retrieval step followed by multilabel immunostaining and examination by confocal microscopy. As examples, we illustrate epitopes localized to the apical and basolateral membranes, and the cytoplasm of enterocytes of normal small intestine and in cases of congenital enteropathies (microvillous inclusion disease and congenital tufting enteropathy). We also demonstrate localization of the bile salt excretion pump protein (BSEP) in bile canalicular membrane of normal hepatocytes and in cases of primary sclerosing cholangitis. To demonstrate colocalization of cytoplasmic and nuclear epitopes we analyzed normal control and hyperplastic pulmonary neuroendocrine cells (PNEC) and neuroepithelial bodies (NEBs), presumed airway sensors in the lungs of infants with bronchopulmonary dysplasia (BPD). As cytoplasmic markers we used anti-bombesin or anti-synaptic vesicle protein 2 (SV2) antibody, respectively, and for nuclear localization, antibodies against neurogenic genes mammalian achaete-scute homolog (Mash1) and prospero homeobox 1 (Prox1), essential for NEB cells differentiation and maturation, hypoxia-inducible factor 1α (HIF1α) a downstream modulator of hypoxia response and a proliferation marker Ki67. The reprobing method consisted of removal of the previously immunolabeled target and immunostaining with different antibodies, facilitating colocalization of enterocyte brush border epitopes as well as HIF1α, Mash1 and Prox1 in PNEC/NEB PNEC and NEBs. As these methods are suitable for routine FFPE pathology samples from various tissues, allowing visualization of multiple epitopes in the same cells/sections with superior contrast and resolution, they are suitable for a wide range of applications in diagnostic pathology and may be particularly well suited for precision medicine diagnostics.

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Innervation of Pulmonary Neuroendocrine Cells and Neuroepithelial Bodies in Developing Rabbit Lung

Cited by 60 publications

References 32 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

Neuro-immune interactions during development of pulmonary fibrosis

Multilabel immunofluorescence and antigen reprobing on formalin-fixed paraffin-embedded sections: novel applications for precision pathology diagnosis

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