Evidence for pleural epithelial-mesenchymal transition in murine compensatory lung growth.

In many mammals, including rodents and humans, removal of one lung results in the compensatory growth of the remaining lung; however, the mechanism of compensatory lung growth is unknown. Here, we investigated the changes in morphology and phenotype of pleural cells after pneumonectomy. Between days 1 and 3 after pneumonectomy, cells expressing α-smooth muscle actin (SMA), a cytoplasmic marker of myofibroblasts, were significantly increased in the pleura compared to surgical controls (p < .01). Scanning electron microscopy of the pleural surface 3 days post-pneumonectomy demonstrated regions of the pleura with morphologic features consistent with epithelial-mesenchymal transition (EMT); namely, cells with disrupted intercellular junctions and an acquired mesenchymal (rounded and fusiform) morphotype. To detect the migration of the transitional pleural cells into the lung, a biotin tracer was used to label the pleural mesothelial cells at the time of surgery. By post-operative day 3, image cytometry of post-pneumonectomy subpleural alveoli demonstrated a 40-fold increase in biotin+ cells relative to pneumonectomy-plus-plombage controls (p < .01). Suggesting a similar origin in space and time, the distribution of cells expressing biotin, SMA, or vimentin demonstrated a strong spatial autocorrelation in the subpleural lung (p < .001). We conclude that post-pneumonectomy compensatory lung growth involves EMT with the migration of transitional mesothelial cells into subpleural alveoli.

Phylogeny and ontogeny of the lymphatic stomata connecting the pleural and peritoneal cavities with the lymphatic system--a review.

This paper reviews the phylogeny and ontogeny of "lymphatic stomata" through which fluids and cells in the pleural and peritoneal cavities enter the lymphatic system. In amphibians, the pleuroperitoneal cavity is connected through numerous pores with the wide subvertebral lymphatic sinus corresponding to the thoracic duct in mammals. In reptiles, direct connections of the pleural and peritoneal cavities with the lymphatic system have been reported. In mammals, the pleural and peritoneal cavities are directly connected with lymphatics through lymphatic stomata. How do lymphatic stomata develop in mammals? In the rat, distinct lymphatics are noted in the subpleural space of the diaphragm periphery in 16 days old embryo. With age, the supleural lymphatics increase and form a polygonal network. They show a tubular appearance and possess many valves. Some of them become endowed with smooth muscle cells. In 19 days old embryos, some lymphatics appear in the subperitoneal space of the diaphragm. They extend centripetally and form many lateral projections that later elongate and connect with those from adjacent lymphatics, thus forming a lattice-like network or "lymphatic lacunae". During early postnatal days, the lymphatic lacunae project many bulges that subsequently come into contact with the pores among mesothelial cells lining the diaphragmatic peritoneum, thus forming lymphatic stomata. They increase until postnatal week 10. The lymphatic stomata in the costal pleura also develop during early postnatal days.

Half-sib analysis of three morphological traits in Drosophila melanogaster under poor nutrition.

Variation in thorax length, wing length and sternopleural bristle number was examined in Drosophila melanogaster reared in stressful and nonstressful environments using paternal half-sib design. Low concentration of yeast in the medium was used as a stress factor. Phenotypic variation of thorax length and wing length was higher under poor nutrition than in the control; in bristle number, phenotypic variation was relatively stable regardless of the environment. Heritability of all the traits analyzed was generally lower under nutritional stress. Heritability changes in thorax length and wing length were mainly due to an increase in the environmental variance under stress, whereas in bristle number, stress resulted in a decrease in genetic variation. Genetic variance in thorax length was higher under poor nutrition; in wing length, no difference in genetic variance between environments was found.

Peritoneal lymphatic stomata of the diaphragm in the mouse: process of their formation.

BACKGROUND: Lymphatic stomata are channels connecting the peritoneal cavity with the lymphatics in the diaphragm. The process of sequential formation of the stomata has not been studied. The objective of this study was to examine the morphogenesis of the lymphatic stomata in mice. METHODS: Ultrathin sections of diaphragms from ddY mice obtained on embryonic day 18 and postnatal days 0, 4, and 10 were observed with a transmission electron microscope. RESULTS: By embryonic day 18 and postnatal day 0, lymphatics were already observed in the submesothelial connective tissue on the peritoneal side of the fetal diaphragm. The lymphatic endothelial cells, but not the mesothelial cells covering the diaphragm, protruded short cytoplasmic processes into the submesothelial connective tissue, and these almost reached the basal surfaces of individual mesothelial cells. By postnatal days 4 and 10, the lymphatic endothelial cells frequently protruded cytoplasmic processes into the submesothelial connective tissue, and the endothelial cell processes broke the continuity of both the basal lamina beneath the mesothelial cells and the submesothelial connective tissue. Neighboring endothelial processes formed a pair of U-shaped folds that were connected with each other via intercellular junctions at the apexes of the U-shaped folds. The disassembly of the intercellular junctions between the U-shaped folds was observed, and the basal surface of the mesothelial cell faced the lymphatic lumen. Dehiscence of the intercellular junctions between the mesothelial cells overlaying the lymphatics was observed, and lymphatic stomata were present. On the pleural side of the diaphragm, lymphatics were already present on embryonic day 18, but it was not observed that the endothelial process spanned the submesothelial connective tissue to the basal surface of the mesothelial cell. CONCLUSIONS: These results suggest the following process of the formation of the lymphatic stomata. (1) Neighboring lymphatic endothelial cells span the submesothelial connective tissue to the basal surfaces of mesothelial cells. (2) The lymphatic stomata are formed by the disassembly of the intercellular junctions between the neighboring endothelial cells and between the mesothelial cells overlying the endothelial cells.

Dopamine-immunoreactive neurones in the central nervous system of the pond snail Lymnaea stagnalis.

The distribution of dopamine and dopamine-immunoreactive neurones was studied in the central nervous system of the snail Lymnaea stagnalis. The results from immunocytochemical labelling were compared with those from the application of the glyoxylic acid fluorescence method and 6-hydroxydopamine-induced pigment labelling. Comparisons were also made between the number of dopamine immunoreactive neurones and the dopamine content of the ganglia, measured by high-performance liquid chromatography. Dopamine immunocytochemistry proved to be superior to the other two histochemical techniques in terms of specificity and sensitivity. The 6-hydroxydopamine-induced pigment labelling failed to prove a useful tool for the in vivo identification of all dopamine-containing neurones. The distribution and number of dopamine-immunoreactive neurones and levels of biochemically measured dopamine in specific ganglia showed a close correspondence. By using the results of the dopamine immunocytochemistry and glyoxylic acid technique, a detailed map of dopamine-containing neurones was constructed. Dopamine-containing inter- and intra-ganglionic axon tracts were also demonstrated. The mapping of dopamine-containing neurones will facilitate further neurophysiological analysis of dopaminergic neural mechanisms in Lymnaea.

The effects of talc pleurodesis on growing swine.

Talc pleurodesis is an effective means of preventing recurrent pneumothorax. We have successfully applied talc pleurodesis with thoracoscopy in children with cystic fibrosis presenting with pneumothorax. However, little is known about the effects of talc pleurodesis on lung compliance in growing children. Therefore, six young pigs (10 weeks old, weighing 15 +/- 1 kg) were prepared for study. In each pig, one hemithorax underwent thoracoscopy and talc pleurodesis (TALC). The other hemithorax underwent either thoracoscopy alone or no procedure (CONTROL). Dynamic and static respiratory mechanics were studied 5 and 10 weeks later. Air flow and airway pressure were measured to calculate dynamic transpulmonary and transrespiratory compliance, and static transpulmonary compliance. At 5 weeks, dynamic transpulmonary and transrespiratory compliance were less in the TALC lungs when compared with CONTROL lungs. At 10 weeks, the differences in dynamic transpulmonary and transrespiratory compliance between the TALC and CONTROL lungs had resolved. Static compliance was lower in the TALC lungs than in the CONTROL lungs at both 5 and 10 weeks, but this was significant only at 10 weeks. There was an improvement in static compliance in the TALC lungs between 5 and 10 weeks, but this only approached significance. At autopsy, there were marked adhesions and pleural thickening in the talc lungs. Histological examination demonstrated no differences in lung parenchyma between the TALC lungs and the CONTROL lungs. Talc pleurodesis causes a temporary impairment in dynamic transpulmonary and transrespiratory compliance that resolves with time and growth. Static compliance is more persistently compromised, but a trend toward improvement with time and growth exists.(ABSTRACT TRUNCATED AT 250 WORDS)

Postnatal development of the respiratory system of the opossum. II. Electron microscopy of the epithelium and pleura.

At birth, the opossum lung is remarkably primitive and consists of a system of branching airways that end in a number of terminal air chambers. From the newborn through the 10 cm stage of development the conducting portion of the lung predominates. The air chambers, which represent portions of the conducting system modified for respiration, are in a constant state of evolution since they are destined to become part of the expanding bronchial system. The airways are devoid of cilia and goblet cells at birth, and are lined by columnar epithelial cells which contain two types of cytoplasmic granules: an electron-dense form and a heterogeneous form. The latter exhibits an electron-dense core surrounded initially by a large halo of flocculent material. This type of granule is not seen beyond the 8 cm stage. The terminal air chambers of the newborn and later stages are lined type I and type II alveolocytes that appear identical to the alveolocytes lining alveoli in the adult. By the 2.5 cm stage, scattered cilia are present in the trachea and bronchi and bands of smooth muscle have differentiated in relation to bronchial epithelium and to proximal areas of the terminal chambers. Citiated cells are separated by ridges composed of light and dark cells which are without cilia and which contain scattered electron-dence granules. Throughout the postnatal period numerous alveolar macrophages and mast cells are noted in relation to the conducting system and pleura. Differentiation of the pleura also occurs during the postnatal period. In the newborn the pleura is simple squamous mesothelium. Later stages develop a thick connective tissue lamina between the pleural mesothelium and lung parenchyma. A large band of elastin is interposed between the mesothelium and underlying bundles of collagen.