Distribution of intracellular and secreted surfactant during postnatal rat lung development (original) (raw)
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Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1988
To clarify perinatal transformations of surfactant we performed lung lavage in term fetuses and in O-24-hold newborn rabbits. Lavage fluid was separated into three pools, namely lavage pellet, lavage supernatant and cells. We found that at birth the pellet contains 94.1 + 1.4% (S.E.) saturated phosphatidylcholine, while the supernatant and cells contain traces of it. At birth the pellet contains secreted lamellar bodies while the supernatant lacks any recognizable structure. After birth, the alveolar saturated phosphatidylcholine level increases 5.1-times in 24 h, the proportions between pools reaching adult values in 90 min (pellet = 75.9 + 4.8%, supernatant = 22.7 * 4.9%), and small vesicles appear in the supernatant, probably originating from the turnover of alveolar surfactant during breathing. The saturated phosphatidylcholine associated with cells remains unchanged. At birth, the 32-38 kDa surfactant apolipoprotein appears to be less extensively sialylated than in adult life.
Analysis of Labeling and Clearance of Lung Surfactant Phospholipids in Rabbit
Journal of Clinical Investigation, 1981
6 h. During 2 h after the application of phospholipids, the radioactivity in the lamellar body fraction increased, and the specific radioactivities approached those in alveolar lavage. The association of phosphatidylglycerol with lamellar bodies was unaffected by myoinositol. Phosphatidylinositol entered more slowly than did phosphatidylglycerol from microsomes to the alveolar lavage fraction, and from alveolar lavage to lamellar bodies. These differences may be ofimportance regarding the poor performance of phosphatidylinositolcontaining surfactant at birth. Further investigations are needed to clarify the possible role for the postulated bidirectional surfactant flux between the lamellar body and alveolar lavage fractions in maintaining the activity of surfactant.
Structure of the phosphatidylcholines of the lung surfactant at birth in normal full term infants
Clinica Chimica Acta, 1976
1. This investigation was undertaken for the purpose of determining the structure of the phosphatidylcholines of lung surfactant system present at birth in normal full term newborn infants. 2. The procedure, using tracheal aspirates as lung secretions, combines a cold-acetone precipitation and a two-dimensional thin-layer chromatography of the lipid extract. 3. Different species of phosphatidylcholines were isolated and found to account together for over 60% of the total phospholipids in tracheal aspirates. Analysis of the fatty acids esterifying the alpha- and beta-carbon of these different phosphatidylcholines showed palmitic acid as the major component with little myristic acid. 4. This fatty acid analysis revealed furthermore that the major phosphatidylcholine fraction was almost exclusively alpha, beta-dipalmitoylphosphatidylcholine. 5. This study shows that the procedure described provides a useful and simple method for the extraction, isolation and characterisation of the functional components of lung surfactant in living human newborns.
Comparative Biochemistry and Physiology Part A: Physiology, 1992
1. We studied the total amount and subcellular distribution of alveolar surfactant, extracted through bronchoalveolar lavage of anesthetized cats and rabbits. This was correlated to several morphometric and ventilatory variables of these animals.2. Lung weight was significantly larger in the cat while respiratory frequency and minute ventilation were significantly larger in the rabbit. No significant differences were observed in tidal volume, total lung capacity, PaO2, PaCO2 and pHa.3. While both species had similar protein contents in the bronchoalveolar lavage, rabbits had larger phospholipid contents, mostly distributed in the lighter, more active subfractions.4. With regard to the estimated values obtained from allometric equations derived for mammals, the rabbit presented a lung weight of nearly one-third of the estimated one, an exceedingly larger minute ventilation (by nearly 60%) and a respiratory frequency twice the calculated one.5. We suggest that the different distribution of alveolar surfactant in these species may be explained by disparities in their ventilatory demands, the rabbit having a higher respiratory frequency and a larger minute ventilation, performed by a mass of lung tissue lower than that corresponding to its body mass.
Developmental regulation of re-uptake of phosphatidylcholine by type II alveolar epithelium
Biochimica et biophysica acta, 1994
Type II alveolar epithelia produce, store and secrete pulmonary surfactant, a phospholipid and protein mixture which stabilizes alveoli at low lung volumes and, thereby, prevents alveolar collapse. We determined the developmental changes in the uptake, metabolism and reutilization of surfactant-related phospholipid in primary cultures of type II cells derived from fetal rat lung. Primary cultures of fetal and neonatal type II cells were incubated in media containing labelled liposomes. After the incubation phospholipids were extracted from the cells and uptake of label was analyzed. Re-uptake of radiolabelled dipalmitoyl phosphatidylcholine (DPPC) was concentration-dependent in undifferentiated fetal cells, differentiated fetal cells and neonatal cells. Re-uptake of DPPC by undifferentiated fetal cells was lower than re-uptake by both differentiated fetal and neonatal cells at 15 and 75 microM PC. Binding of DPPC to the cell surface involved a protein interaction, since trypsin was ...
Development of the pulmonary surfactant system in the amniotes
Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 2000
Pulmonary surfactant (PS) is a complex mixture of phospholipids, neutral lipids and proteins that lines the inner surface of the lung. Here, it modulates surface tension thereby increasing lung compliance and preventing the transudation of fluid. In mammals, the PS system develops towards the end of gestation, characterized by an increase in the saturation of phospholipids in lung washings and the appearance of surfactant proteins in amniotic fluid. Birth, the transition from in utero to the external environment, is a rapid process. At this time, the PS system is important in opening and clearing the lung of fluid in order to initiate pulmonary ventilation. In oviparous vertebrates, escape from an egg can be a long and exhausting process. The young commence pulmonary ventilation and hatching by 'pipping' through the eggshell, where they remain for some time, presumably clearing their lungs. This paper relates changes in the development of the pulmonary surfactant system within the non-mammalian amniotes in response to birth strategy, lung morphology and phylogeny in order to determine the conservatism of this developmental process. Total phospholipid (PL), disaturated phospholipid (DSP) and cholesterol (Chol) were quantified from lung washings of embryonic and hatchling chickens, bearded dragons (oviparous), sleepy lizards (viviparous), snapping turtles and green sea turtles throughout the final stages of incubation and gestation. In all cases, the pattern of development of the pulmonary surfactant lipids was consistent with that of mammals. PL and DSP increased throughout the latter stages of development and Chol was differentially regulated from the PLs. Maximal secretion of both PL and DSP occurred at 'pipping' in oviparous reptiles, coincident with the onset of airbreathing. Similarly, the amount of DSP relative to total PL was maximal immediately after the initiation of airbreathing in chickens. The relative timing of the appearance of the lipids differed between groups. In the oviparous lizard, surfactant lipids were released over a relatively shorter time than that of the sleepy lizard, turtles, birds and mammals. Thus, despite temporal differences and vastly different lung morphologies, birth strategies and phylogenies, the overall development and maturation of the PS system is highly conserved amongst the amniotes.
Histochemistry and Cell Biology, 2005
Surfactant proteins (SP) have an important impact on the function of the pulmonary surfactant. In contrast to humans, rat lungs are immature at birth. Alveolarization starts on postnatal day 4. Little is known about the distribution of SP during postnatal alveolarization. By immunoelectron microscopy, we studied the distribution of SPA , SP-D, SP-B, and precursors of SP-C in type II pneumocytes before, near the end and after alveolarization and in mature lungs. We determined the subcellular volume fractions and the relative labeling index to obtain information about preferential labeling of compartments and non-randomness of labeling. Independently of alveolarization, the overall cellular distribution of SP was non-random. A preferential labeling for SPA and SP-D was found in small vesicles and multivesicular bodies (mvb). SP-B and precursors of SP-C were localized in mvb and lamellar bodies (lb). There are no postnatal changes in labeling for all three SP in these compartments. Labeling intensity for SP-B in lb increased in close correlation with a significant increase in the volume fractions of lb during alveolarization. Our results support the concept that postnatal alveolarization in rat lungs is associated with significant increases in the SP-B content in lb and volume fraction of lb in type II pneumocytes. The postnatal compartment-specific distribution of SPA , precursors of SP-C and SP-D does not change.
Phospholipid Metabolism in Lung Surfactant
Membrane Dynamics and Domains, 2004
Pulmonary surfactant is a mixture of lipids, mostly phospholipids, and proteins that allows for breathing with minimal effort. The current chapter discusses the metabolism of the phospholipids of this material. Surfactant phospholipids are synthesized in the type II epithelial cells of the lung. The lipids and surfactant proteins are assembled in intracellular storage organelles, called lamellar bodies, and are subsequently secreted into the alveolar space. Within this extracellular space surfactant undergoes several transformations. First the lamellar bodies unravel to form a highly organized lattice-like lipid: protein structure tubular myelin. Second, the organized structures, in particular tubular myelin, adsorb to form a lipid at the air-liquid interface of the alveoli. It is, in fact, this surface tension reducing film that is responsible for the physiological role of surfactant, to prevent lung collapse and allow ease of inflation. Third, the surface film is converted to a small vesicular form. Finally, these small vesicles are taken-up by the type II cells for recycling and degradation and by alveolar macrophages for degradation.
Phosphatidylcholine Molecular Species in Lung Surfactant
American Journal of Respiratory Cell and Molecular Biology, 2001
Surfactant reduces surface tension at the air-liquid interface of lung alveoli. While dipalmitoylphosphatidylcholine (PC16:0/ 16:0) is its main component, proteins and other phospholipids contribute to the dynamic properties and homeostasis of alveolar surfactant. Among these components are significant amounts of palmitoylmyristoylphosphatidylcholine (PC16:0/ 14:0) and palmitoylpalmitoleoylphosphatidylcholine (PC16:0/ 16:1), whereas in surfactant from the rigid tubular bird lung, PC16:0/14:0 is absent and PC16:0/16:1 strongly diminished. We therefore hypothesized that the concentrations of PC16:0/14:0 and PC16:0/16:1 in surfactants correlate with differences in the respiratory physiology of mammalian species. In surfactants from newborn and adult mice, rats, and pigs, molar fractions of PC16:0/14:0 and PC16:0/16:1 correlated with respiratory rate. Labeling experiments with [methyl-3 H]choline in mice and perfused rat lungs demonstrated identical alveolar proportions of total and newly synthesized PC16:0/14:0, PC16:0/16:1, and PC16:0/16:0, which were much higher than those of other phosphatidylcholine species. In surfactant from human term and preterm neonates, fractional concentrations not only of PC16:0/16:0 but also of PC16:0/14:0 and PC16:0/ 16:1 increased with maturation. Our data emphasize that PC16:0/14:0 and PC16:0/16:1 may be important surfactant components in alveolar lungs, and that their concentrations are adapted to respiratory physiology.