Contractile Interstitial Cells Research Articles

Metabolism of Vitamin A Affected by Prostaglandin F Synthase in Contractile Interstitial Cells of Bovine Lung

Contractile interstitial cells (CIC), the major component of the alveolar septum of the bovine lung are enriched in prostaglandin (PG) F synthase (Fukui, M., Fujimoto, T., Watanabe, K., Endo, K., and Kuno, K. (1996) J. Histochem. Cytochem. 44, 251–257.). The enzyme catalyzes not only the reduction of PGD2 and PGH2 but also that of various carbonyl compounds (Watanabe, K., Yoshida, R., Shimizu, T., and Hayaishi, O. (1985) J. Biol. Chem. 260, 7035–7041). Here, we report that retinal (vitamin A-aldehyde) was reduced to retinol (vitamin A-alcohol) dose- and time-dependently by PGF synthase using NADPH as a cofactor. The Km value of PGF synthase for retinal was about 20 μM, a same order to that for PGH2. The conversion of retinal to retinol was also observed in cultured CIC, as demonstrated by the greenish fluorescence characteristic of retinol. Thus, retinal might be one of the natural substrates for PGF synthase in vivo, and retinol synthesized from retinal in CIC may play physiological and pathological roles in the lungs.

The pulmonary lipofibroblast (lipid interstitial cell) and its contributions to alveolar development.

The pulmonary lipofibroblast is located in the alveolar interstitium and is recognizable by its characteristic lipid droplets. During alveolar development it participates in the synthesis of extracellular matrix structural proteins, such as collagen and elastin, and as an accessory cell to the type II pneumocyte, in the synthesis of surfactant. The lipofibroblast contains cortical contractile filaments and is thereby related to the contractile interstitial cells that are normally found at the alveolar septal tips and after lung injury. The morphologic, immunologic, and biochemical characteristics of the lipofibroblast and its probable physiologic functions are reviewed. The retinoid and lipid metabolism of the lipofibroblast is compared with that of the hepatic lipocyte and the adipocyte. Although the functions of the lipofibroblast remain incompletely characterized, this cell type is emerging as an important contributor to pulmonary alveolar septal development.

Alterations of lung structure in experimental diabetes, and diabetes associated with hyperlipidaemia in hamsters.

Since hyperglycaemia is known to affect normal pulmonary physiology and biochemistry and few structure-function correlations have been reported, we designed experiments on hamsters subjected to streptozotocin-induced diabetes or diabetes associated with hyperlipidaemia, and investigated the impact of these conditions on the lung structure. At time intervals ranging 2-24 weeks from the inception of disease (without correcting blood glucose with insulin), the animals were sacrificed, and plasma glucose and cholesterol assayed. The lung was processed for electron microscopy, and the structural changes of the capillary and venular endothelium, of epithelial cells, and interstitium were examined. In diabetic animals, especially after 6 weeks of disease, a gradual narrowing of approximately 35% of the capillaries and approximately 30% of the alveoli, and hyperplasia of the extracellular matrix, rich in collagen bundles, were observed. Frequently, capillaries contained adherent intravascular macrophages suggestive of an inflammatory process. The capillary endothelium was characterized by numerous plasmalemmal vesicles, often fused, well-developed synthesizing apparatus (endoplasmic reticulum and Golgi complex) and cytoskeleton, and an uneven distribution of the anionic sites on the luminal plasmalemma. The venular endothelium was particularly rich in Weibel-Palade bodies. The alveolar epithelium was often collapsed, compressing surfactant within the airspace. The lung interstitium was apparently enlarged, and the fibroblasts and contractile interstitial cells frequently contained lipid droplets. These alterations were more pronounced and occurred at a faster rate (4 weeks) in diabetes associated with hyperlipidaemia. The structural modifications reported in this study support the functional disturbances observed in association with hyperglycaemia, sustaining the conclusion that the lung is an organ affected by diabetes.

Hypoxic contraction of contractile interstitial cells isolated from bovine lung.

Although contractile interstitial cells (CIC) in the alveolar septum have been suggested to be involved in hypoxic pulmonary vasoconstriction (HPV), direct demonstration of cellular contraction under hypoxia has been lacking. To achieve this, we purified CIC from collagenase-dissociated bovine lung cells and examined the response of these cells to hypoxia. Prostaglandin (PG) F synthase served as a marker of CIC, and the isolated PGF synthase-positive cells were shown to preserve the ultrastructural features characteristic of CIC, most notably bundles of microfilaments. Isolated CIC seeded onto collagen gel disks became embedded and formed a lattice network with collagen fibrils. Exposure of these CIC-bearing gels to hypoxia (PO2 = 20-40 Torr) evoked a reversible reduction in gel volume, as assessed by measuring the surface area of the gel disks photographically. Thus CIC were shown to contract under hypoxia, providing the supportive evidence for the involvement of CIC in HPV.

Prostaglandin F synthase is localized to contractile interstitial cells in bovine lung.

It was recently found that certain cells in the alveolar septum of the bovine lung are enriched in prostaglandin F (PGF) synthase. In this study we used immunohistochemical techniques at both light and electron microscopic levels to further characterize the PGF synthase-positive cells. By double immunofluorescence staining of bovine lung cryostat sections, the alveolar septal cells labeled by anti-PGF synthase antibody were also intensely labeled for cytoplasmic actin but not for alpha-smooth muscle actin. This labeling pattern suggests that the PGF synthase-positive cells in the septum are "contractile interstitial cells," which resemble conventional fibroblasts but characteristically contain prominent bundles of actin filaments. Immunogold electron microscopy of ultra-thin frozen sections of bovine lung showed that alveolar interstitial cells extending long cytoplasmic processes and closely associated with alveolar capillaries were intensely labeled for PGF synthase. Capillary endothelial cells, alveolar epithelial cells, and some fibroblastic cells were devoid of labeling. On the basis of these findings, we conclude that PGF synthase is specifically expressed in contractile interstitial cells within the alveolar septum. The protein may be a useful marker for contractile interstitial cells, whose physiological function and role in various pathological conditions have not been characterized in detail.

Hypoxia-evoked contraction of contractile interstitial cells isolated from bovine lung/possible mechanism of hypoxic pulmonary vasoconstriction

Hypoxia-evoked contraction of contractile interstitial cells isolated from bovine lung/possible mechanism of hypoxic pulmonary vasoconstriction

Cytoskeletal features of alveolar myofibroblasts and pericytes in normal human and rat lung.

Frozen or paraffin-embedded human and rat lung specimens were stained with antibodies against total actin, alpha-smooth muscle (SM) actin, vimentin, desmin, or gelsolin. Alveolar interstitial myofibroblasts [i.e., contractile interstitial cells (CIC)] were labeled by total actin antibody but not by alpha-SM actin antibody. They stained for vimentin and gelsolin and, in rat lungs, most of them for desmin. Pericytes located around venules at the junction of three alveolar septa were always positive for alpha-SM actin and never for desmin. Tissue samples were also immunostained by an alpha-SM actin antibody and studied by electron microscopy. With this technique we confirmed that cells, identified as pericytes on the basis of their location, were intensely labeled by alpha-SM actin antibodies, whereas alveolar myofibroblasts were not. We conclude that in the lung interstitium pericytes and alveolar myofibroblasts have distinct cytoskeletal features, alpha-SM actin antibody staining being a simple method to distinguish between them. Furthermore, it appears that alveolar myofibroblasts have a peculiar pattern of cytoskeletal protein composition which, in the rat, is similar to that previously described for stromal cells in uterine submucosa, liver sinusoids (Ito cells), or the core of intestinal villi.

Orientation of Adhering Junctions between Bovine Pulmonary Fibroblasts

The ultrastructure and orientation of microfilament-attaching junctions between pulmonary fibroblasts (also known as myofibroblasts or contractile interstitial cells) of bovine lung septa were investigated by transmission electron microscopy. Adhering junctions similar to the fascia adherens of the intercalated disc, but of a macular design, link microfilament bundles of adjacent fibroblasts. The bundles of microfilaments joined by junctions were usually aligned perpendicularly to the axis of the alveolar wall. Gap junctions were located in close proximity to the adhering junctions, presumably to co-ordinate the contraction of the cells. The data indicate that fibroblasts are able to form multicellular contractile units within healthy, mature lung parenchyma.

Cellular alterations in the alveolar wall in bleomycin-induced pulmonary fibrosis in rats. An ultrastructural morphometric study.

This study was designed to measure quantitatively changes in populations of major alveolar cell types, with a particular focus on nonmuscle, contractile interstitial cells, in lungs of rats 14 and 28 days after a single intratracheal instillation of bleomycin sulfate. Standard ultrastructural morphometric techniques, including stratified random sampling, were applied to airway-perfused lungs from bleomycin-instilled and control (saline-instilled) rats. Cell numbers and volumes were calculated for each major cell type populating the pulmonary parenchyma. At 14 days postinstillation, the number and volume of fibroblasts, contractile interstitial cells, Type II epithelial cells, and macrophages were increased significantly. At 28 days, the largest increase was in contractile interstitial cells, approximately tenfold in number and fourfold in volume over control cells. The number and volume of non-filament-containing fibroblasts did not differ between control and treated groups at this time. These data suggest proliferating fibroblasts undergo "differentiation" into cell types with increasing contractile capability during the development of fibrosis in this animal model. This study represents the first quantitative description of increases in contractile interstitial cells related to pulmonary fibrosis.

Actin in peripheral rat lung: S1 labeling and structural changes induced by cytochalasin.

Actin was identified with S1-labeling in type I and II cells, pericytes, and capillary endothelial cells in the peripheral lung of the adult rat. In type II cells abundant actin was present in the apex of cells, in microvilli, and in close association with lamellar bodies near the cell surface. Lamellar bodies secreting their content into the lumen of alveoli were always surrounded by a thick layer of actin-like material. In specimens treated with cytochalasin D the surface of type I, type II and contractile interstitial cells became irregular. In type II cells, lamellar bodies were no longer surrounded by actin-like material and no exocytic profiles of lamellar bodies were encountered. In type II cells actin filaments may be involved in moving lamellar bodies through the cytoplasm and/or in their secretion into alveoli. These observations also suggest that intact actin filaments may be required for maintenance of cell shape in various lung cells and that cells containing actin may be capable of limited contraction.

Pulmonary α-adrenoceptors: Autoradiographic localization using [ 3H]prazosin

We determined the distribution of pulmonary α-adrenoceptors by autoradiographic localisation of [ 3H]prazosin binding to frozen sections of ferret lung. Specific binding of [ 3H]prazosin to lung sections was saturable and of high affinity (K D = 0.44 ± 0.55 nM; mean ± S.E., n = 5), with a specificity indicating binding to α 1-receptors. Autoradiographic showed that α 1-receptors were present in highest density in vascular smooth muscle (small vessels > large vessels), and were also present in airway submucosal glands and epithelium. There was also scantly labelling of alveolar walls which may be to contractile interstitial cells (Kapanci cells). Although smooth muscle of bronchi showed little labelling, surprisingly that of bronchioles was heavily labelled. The high density of α-receptors in small airways may be relevant to asthma in which α-adrenergic responses are activated. This method offers a means by which autonomic receptors of small airways may be investigated without the confusing contribution of other contractile elements and larger airways.

Role of histamine in the hypoxic pulmonary artery vasoconstriction

This gives support to the hypothesis ofKapanci et al. [12], suggesting a direct action of hypoxia on contractile interstitial cells in pulmonary alveolar septa without release of any mediators.