Heterogeneity in the immunolocalization of cytokeratin-specific monoclonal antibodies in the rat lung: evaluation of three different alveolar epithelial cell types

Kasper, Michael; Rudolf, Thomas; Verhofstad, A.A.J.; Schuh, Dieter; Müller, Martin

doi:10.1007/bf00268879

Cited by 38 publications

(36 citation statements)

References 24 publications

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“…The antibody MNF-116 recognizes a wide range of cytokeratins (21). In lung parenchyma it binds specifically to the apical surface of type II epithelial cells (13). Two cells that were positive for both labels are shown in Fig.…”

Section: Morphologymentioning

confidence: 99%

Suppression of cell proliferation and programmed cell death by dexamethasone during postnatal lung development

Luyet

Burri

Schittny

2002

American Journal of Physiology-Lung Cellular and Molecular Phys

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Prematurely born babies are often treated with glucocorticoids. We studied the consequences of an early postnatal and short dexamethasone treatment (0.1–0.01 μg/g, days 1–4) on lung development in rats, focusing on its influence on peaks of cell proliferation around day 4 and of programmed cell death at days 19–21. By morphological criteria, we observed a dexamethasone-induced premature maturation of the septa ( day 4), followed by a transient septal immatureness and delayed alveolarization leading to complete rescue of the structural changes. The numbers of proliferating (anti-Ki67) and dying cells (TdT-mediated dUTP nick end labeling) were determined and compared with controls. In dexamethasone-treated animals, both the peak of cell proliferation and the peak of programmed cell death were reduced to baseline, whereas the expression of tissue transglutaminase (transglutaminase-C), another marker for postnatal lung maturation, was not significantly altered. We hypothesize that a short neonatal course of dexamethasone leads to severe but transient structural changes of the lung parenchyma and influences the balance between cell proliferation and cell death even in later stages of lung maturation.

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Section: Morphologymentioning

confidence: 99%

Suppression of cell proliferation and programmed cell death by dexamethasone during postnatal lung development

Luyet

Burri

Schittny

2002

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…An additional antibody was used to identify cytokeratins, which are cytoskeletal components of airway epithelial cells, to visualize the cytoskeleton and localize ubiquitin within the cytoskeleton. 45 Primary antibodies (Table 3 45 ) were commercially available and tested/approved for immunohistochemistry by the manufacturer. Three different antibodies were used to detect antigens important in autophagy of misfolded proteins.…”

Section: Fluorescence Assaysmentioning

confidence: 99%

“…Product details are listed in Table 3. 45 Remaining antibodies were used to localize total ubiquitin, K63-ubiquitin, and SQSTM1 to specific cellular and subcellular sites. In addition, we correlated their presence with diacetyl-induced morphological changes or changes in RNA expression in the lung, nose, and brain.…”

Section: Fluorescence Assaysmentioning

confidence: 99%

Accumulation of Ubiquitin and Sequestosome-1 Implicate Protein Damage in Diacetyl-Induced Cytotoxicity

Hubbs

Fluharty

Edwards

et al. 2016

The American Journal of Pathology

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Inhaled diacetyl vapors are associated with flavorings-related lung disease, a potentially fatal airway disease. The reactive a-dicarbonyl group in diacetyl causes protein damage in vitro. Dicarbonyl/ L-xylulose reductase (DCXR) metabolizes diacetyl into acetoin, which lacks this a-dicarbonyl group. To investigate the hypothesis that flavorings-related lung disease is caused by in vivo protein damage, we correlated diacetyl-induced airway damage in mice with immunofluorescence for markers of protein turnover and autophagy. Western immunoblots identified shifts in ubiquitin pools. Diacetyl inhalation caused dose-dependent increases in bronchial epithelial cells with puncta of both total ubiquitin and K63-ubiquitin, central mediators of protein turnover. This response was greater in Dcxr-knockout mice than in wild-type controls inhaling 200 ppm diacetyl, further implicating the a-dicarbonyl group in protein damage. Western immunoblots demonstrated decreased free ubiquitin in airway-enriched fractions. Transmission electron microscopy and colocalization of ubiquitin-positive puncta with lysosomal-associated membrane proteins 1 and 2 and with the multifunctional scaffolding protein sequestosome-1 (SQSTM1/p62) confirmed autophagy. Surprisingly, immunoreactive SQSTM1 also accumulated in the olfactory bulb of the brain. Olfactory bulb SQSTM1 often congregated in activated microglial cells that also contained olfactory marker protein, indicating neuronophagia within the olfactory bulb. This suggests the possibility that SQSTM1 or damaged proteins may be transported from the nose to the brain. Together, these findings strongly implicate widespread protein damage in the etiology of flavorings-related lung disease. (Am J Pathol 2016, 186: 2887e2908; http://dx

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“…The first section was pretreated with pepsin and stained with a monoclonal mouse antibody MNF-116 (Dakopatts, Gostrup, Denmark), which recognizes type II epithelial cells (4,18). Briefly, the sections were blocked with Tris-buffered saline (TBS) containing 1 mg/mL of BSA (TBS/BSA) for 30 min and incubated with the antibody, diluted 1:10 in TBS, for 30 min.…”

Section: Identification Of Apoptosis In the Epithelial Cellsmentioning

confidence: 99%

Angiotensin II Receptor Blockade Inhibits Pneumocyte Apoptosis in Experimental Meconium Aspiration

et al. 2004

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Lung tissue inflammation and apoptosis are implicated in the pathogenesis of meconium aspiration-induced lung injury in the newborn, but the mechanisms of these reactions are still poorly known. We investigated the time-dependent leukocyte influx and appearance of apoptosis, as well as the contribution of angiotensin (ANG) II receptor action on these processes in the meconium-induced lung injury. Experimental meconium aspiration was induced by intratracheal instillation of human meconium in 18 rats, and eight rats were further pretreated with an unspecific ANG II receptor inhibitor saralasin. Rats were ventilated with 60% oxygen for 1, 3, or 5 h, and the lungs were then studied histologically for tissue injury and with DNA nick-end labeling and electron microscopy for apoptotic cell death. Lung tissue myeloperoxidase activity and expression of angiotensinogen mRNA and endothelial monocyte-activating polypeptide (EMAP) II protein were also analyzed. The meconium-instilled lungs showed increasing neutrophil migration and histologic injury after the first hour, whereas the number of epithelial apoptotic cells was elevated from the control level throughout the study. Myeloperoxidase activity was high, and the angiotensinogen mRNA and EMAP II protein was up-regulated at 5 h after the meconium insult. Pretreatment with saralasin significantly prevented the increase in lung tissue myeloperoxidase activity, EMAP II, and lung epithelial apoptosis. The results suggest that pulmonary meconium insult rapidly results in epithelial apoptosis, before significant neutrophil sequestration into the lungs. Perinatal aspiration of meconium frequently results in severe pulmonary failure with ventilation-perfusion mismatching in the lungs, hypoxemia, and increase in pulmonary vascular resistance, associated with high morbidity and mortality in fullor postterm newborn infants. The pathophysiology of the neonatal meconium aspiration syndrome (MAS) is complex, but inflammation with accumulation of polymorphonuclear leukocytes in the pulmonary tissue is believed to be a central event in the development of acute tissue damage (1, 2). This inflammatory reaction is associated with increased pulmonary vascular permeability leading to proteinaceous exudation into the alveolar spaces and inactivation of the pulmonary surfactant and may together with direct toxic effects of meconium significantly contribute to the lung injury process (1, 2). Still, the clinical effects of anti-inflammatory therapeutic approaches and surfactant replacement treatment have often been unsatisfactory and have not consistently improved the outcome of severe complications of this disease (1-3). New advances in the pathophysiology of MAS therefore are needed.Apoptosis, programmed cell death, is usually regarded as a physiologic mechanism in organ remodeling (4) but has been

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Heterogeneity in the immunolocalization of cytokeratin-specific monoclonal antibodies in the rat lung: evaluation of three different alveolar epithelial cell types

Cited by 38 publications

References 24 publications

Suppression of cell proliferation and programmed cell death by dexamethasone during postnatal lung development

Suppression of cell proliferation and programmed cell death by dexamethasone during postnatal lung development

Accumulation of Ubiquitin and Sequestosome-1 Implicate Protein Damage in Diacetyl-Induced Cytotoxicity

Angiotensin II Receptor Blockade Inhibits Pneumocyte Apoptosis in Experimental Meconium Aspiration

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