Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1987 Apr;79(4):1133–1138. doi: 10.1172/JCI112929

Degradation of dipalmitoyl phosphatidylcholine by isolated rat granular pneumocytes and reutilization for surfactant synthesis.

A Chander, J Reicherter, A B Fisher
PMCID: PMC424294  PMID: 3558819

Abstract

We investigated metabolic utilization of exogenous (modelled after lung surfactant) phospholipids by granular pneumocytes in primary culture. Cells were incubated for 21, 65, and 140 min with [3H-methyl]dipalmitoylphosphatidylcholine (DPPC) containing liposomes prepared from synthetic lipids. Radioactivity in cellular phosphatidylcholine (PC) declined steadily to 50% of the total trypsin-resistant cell-associated radioactivity. The proportion of radioactivity increased with time in cytidine-5'-diphosphate-choline and phosphorylcholine, which suggested reutilization of choline for PC synthesis. Cells incubated with liposomes for 2 h revealed that of the total cell-associated radioactivity, 7% was in lamellar bodies and 10% in the microsomal fraction. The lipid-associated radioactivity was 24% in "soluble," 96% in lamellar bodies, and 92% in the microsomal fraction. Percent of total PC label recovered in disaturated PC of microsomal fractions decreased (slope = -5.27%/h) with time of incubation (r = 0.67). Incubation of cells with liposomes containing ([3H-methyl]choline-[14C]palmitoyl) DPPC led to altered isotope ratios in both lamellar bodies and microsomes. These observations indicate that granular pneumocytes degrade exogenous PC and resynthesize PC from degradation products.

Full text

PDF
1135

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Brown L. A., Longmore W. J. Adrenergic and cholinergic regulation of lung surfactant secretion in the isolated perfused rat lung and in the alveolar type II cell in culture. J Biol Chem. 1981 Jan 10;256(1):66–72. [PubMed] [Google Scholar]
  4. Chander A., Claypool W. D., Jr, Strauss J. F., 3rd, Fisher A. B. Uptake of liposomal phosphatidylcholine by granular pneumocytes in primary culture. Am J Physiol. 1983 Nov;245(5 Pt 1):C397–C404. doi: 10.1152/ajpcell.1983.245.5.C397. [DOI] [PubMed] [Google Scholar]
  5. Chander A., Dodia C. R., Gil J., Fisher A. B. Isolation of lamellar bodies from rat granular pneumocytes in primary culture. Biochim Biophys Acta. 1983 Aug 29;753(1):119–129. doi: 10.1016/0005-2760(83)90105-4. [DOI] [PubMed] [Google Scholar]
  6. Chander A., Fisher A. B., Strauss J. F., 3rd Role of an acidic compartment in synthesis of disaturated phosphatidylcholine by rat granular pneumocytes. Biochem J. 1982 Dec 15;208(3):651–658. doi: 10.1042/bj2080651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Desai R., Tetley T. D., Curtis C. G., Powell G. M., Richards R. J. Studies on the fate of pulmonary surfactant in the lung. Biochem J. 1978 Nov 15;176(2):455–462. doi: 10.1042/bj1760455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dobbs L. G., Mason R. J. Pulmonary alveolar type II cells isolated from rats. Release of phosphatidylcholine in response to beta-adrenergic stimulation. J Clin Invest. 1979 Mar;63(3):378–387. doi: 10.1172/JCI109313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fisher A. B., Dodia C., Chander A. Beta-adrenergic mediators increase pulmonary retention of instilled phospholipids. J Appl Physiol (1985) 1985 Sep;59(3):743–748. doi: 10.1152/jappl.1985.59.3.743. [DOI] [PubMed] [Google Scholar]
  10. Fisher A. B., Furia L., Berman H. Metabolism of rat granular pneumocytes isolated in primary culture. J Appl Physiol Respir Environ Exerc Physiol. 1980 Oct;49(4):743–750. doi: 10.1152/jappl.1980.49.4.743. [DOI] [PubMed] [Google Scholar]
  11. Goerke J. Lung surfactant. Biochim Biophys Acta. 1974 Dec 16;344(3-4):241–261. doi: 10.1016/0304-4157(74)90009-4. [DOI] [PubMed] [Google Scholar]
  12. Hallman M., Epstein B. L., Gluck L. Analysis of labeling and clearance of lung surfactant phospholipids in rabbit. Evidence of bidirectional surfactant flux between lamellar bodies and alveolar lavage. J Clin Invest. 1981 Sep;68(3):742–751. doi: 10.1172/JCI110310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Infante J. P. Impaired glycerophosphorylcholine synthesis in murine muscular dystrophy. Med Biol. 1985;63(2):81–87. [PubMed] [Google Scholar]
  14. Jacobs H., Jobe A., Ikegami M., Conaway D. The significance of reutilization of surfactant phosphatidylcholine. J Biol Chem. 1983 Apr 10;258(7):4159–4165. [PubMed] [Google Scholar]
  15. Jacobs H., Jobe A., Ikegami M., Miller D., Jones S. Reutilization of phosphatidylcholine analogues by the pulmonary surfactant system. The lack of specificity. Biochim Biophys Acta. 1984 Apr 18;793(2):300–309. doi: 10.1016/0005-2760(84)90333-3. [DOI] [PubMed] [Google Scholar]
  16. Jobe A., Kirkpatrick E., Gluck L. Labeling of phospholipids in the surfactant and subcellular fractions of rabbit lung. J Biol Chem. 1978 Jun 10;253(11):3810–3816. [PubMed] [Google Scholar]
  17. Jobe A. The labeling and biological half-life of phosphatidylcholine in subcellular fractions of rabbit lung. Biochim Biophys Acta. 1977 Dec 21;489(3):440–453. doi: 10.1016/0005-2760(77)90165-5. [DOI] [PubMed] [Google Scholar]
  18. King R. J., Martin H. M. Effects of inhibiting protein synthesis on the secretion of surfactant by type II cells in primary culture. Biochim Biophys Acta. 1981 Jan 26;663(1):289–301. doi: 10.1016/0005-2760(81)90215-0. [DOI] [PubMed] [Google Scholar]
  19. King R. J., Martin H. Intracellular metabolism of the apoproteins of pulmonary surfactant in rat lung. J Appl Physiol Respir Environ Exerc Physiol. 1980 May;48(5):812–820. doi: 10.1152/jappl.1980.48.5.812. [DOI] [PubMed] [Google Scholar]
  20. King R. J. Utilization of alveolar epithelial type II cells for the study of pulmonary surfactant. Fed Proc. 1979 Nov;38(12):2637–2643. [PubMed] [Google Scholar]
  21. Mason R. J., Nellenbogen J., Clements J. A. Isolation of disaturated phosphatidylcholine with osmium tetroxide. J Lipid Res. 1976 May;17(3):281–284. [PubMed] [Google Scholar]
  22. Mitnick M., DeMarco B., Gibbons J. M. Amniotic fluid phosphatidylglycerol and phosphatidylinositol separated by stepwise-development thin-layer chromatography. Clin Chem. 1980 Feb;26(2):277–281. [PubMed] [Google Scholar]
  23. Nagley P., Hallinan T. The use of radioactive choline as a label for microsomal membranes. I. Selectivity of label for endoplasmic reticulum and specificity for lecithin. Biochim Biophys Acta. 1968 Sep 17;163(2):218–225. doi: 10.1016/0005-2736(68)90100-4. [DOI] [PubMed] [Google Scholar]
  24. Robinson B. S., Snoswell A. M., Runciman W. B., Upton R. N. Uptake and output of various forms of choline by organs of the conscious chronically catheterized sheep. Biochem J. 1984 Jan 15;217(2):399–408. doi: 10.1042/bj2170399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sleight R. G., Pagano R. E. Transport of a fluorescent phosphatidylcholine analog from the plasma membrane to the Golgi apparatus. J Cell Biol. 1984 Aug;99(2):742–751. doi: 10.1083/jcb.99.2.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Spitzer H. L., Norman J. R., Morrison K. In vivo studies of [Me-3H]choline and [1,2-14C2]choline incorporation into lung and liver lecithins. Biochim Biophys Acta. 1969 Apr 29;176(3):584–590. doi: 10.1016/0005-2760(69)90224-0. [DOI] [PubMed] [Google Scholar]
  27. Yavin E. Regulation of phospholipid metabolism in differentiating cells from rat brain cerebral hemispheres in culture. Patterns of acetylcholine phosphocholine, and choline phosphoglycerides labeling from (methyl-14C)choline. J Biol Chem. 1976 Mar 10;251(5):1392–1397. [PubMed] [Google Scholar]
  28. de Duve C., de Barsy T., Poole B., Trouet A., Tulkens P., Van Hoof F. Commentary. Lysosomotropic agents. Biochem Pharmacol. 1974 Sep 15;23(18):2495–2531. doi: 10.1016/0006-2952(74)90174-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES