, Volume 85, Issue 6, pp 497–504 | Cite as

Histochemical characterization of an antigen specific for the great alveolar cell in the mouse lung

  • F. J. Van Hemert
  • A. A. W. Ten Have-Opbroek
  • C. J. M. Otto-Verberne


Previous papers reported on a specific antigenic marker for the great alveolar (type-II) cell of the mouse lung and described its recognition by a specific rabbit antiadult mouse lung serum. In the present study light- and electron-microscopical immunohistochemistry on fixed mouse lung sections showed the presence of the marker on the alveolar surface. The antigenic determinants recognized by the antibody were further characterized by immunoblotting and immunoprecipitation studies after in vitro translation of mouse lung messenger RNA.

Immunoblots of a surfactant-enriched pellet of a bronchoalveolar lavage fraction of mouse lung showed that the antibody reacted with surfactant-associated proteins having apparent molecular weights of about 27,000, 32,000, and 38,000 daltons in SDS gels. Immunoblots of mouse-lung homogenate revealed the presence of 27,000, 30,000, 39,000, and 41,000 dalton proteins, presumably also surfactant-associated proteins. Immunoprecipitation after in vitro translation of mouse-lung mRNA showed specific reactivity only with a 12,000 dalton polypeptide, a component of the cell marker we were unable to relate to surfactant. Our findings indicate that the 12,000 dalton component of the antigenic marker for the great alveolar cell is a polypeptide whose synthesis is a lung-specific process and that the immunoreaction of the larger and surfactant-associated components is due to post-translational modifications.


Mouse Lung Apparent Molecular Weight Antigenic Determinant Lung Section Alveolar Surface 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Arvidson S, Holme T, Wadström T (1971) Influence of cultivation conditions on the production of extracellular proteins by Staphylococcus aureus. Acta Pathol Microbiol Scand Sect B 79:399–405Google Scholar
  2. Auffray C, Rougeon F (1980) Purification of mouse immunoglobulin heavy chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem 107:303–314Google Scholar
  3. Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acidcellulose. Proc Natl Acad Sci USA 69:1408–1412Google Scholar
  4. Benson BJ, Williams MC, Hawgood S, Sargeant T (1984) Role of lung surfactant-specific proteins in surfactant structure and function. In: Wichert E von (ed) Current concepts in surfactant research. Prog Respir Res 18:83–92Google Scholar
  5. Bhattacharyya SN, Passero MA, DiAugustine RP, Lynn WS (1975) Isolation and characterization of two hydroxyproline-containing glycoproteins from normal animal lung lavage and lamellar bodies. J Clin Invest 55:914–920Google Scholar
  6. Claypool WD, Wang DL, Chander A, Fisher AB (1984) “Hydrophobic” surfactant apoproteins and augmentation of phospholipid recycling. Exp Lung Res 6:215–222Google Scholar
  7. Clements JA, King RJ (1976) Composition of surface active material. In: Crystal RG (ed) The biochemical basis of pulmonary function. Marcel Dekker, New York, pp 363–387Google Scholar
  8. Floros J, Phelps DS, Taeusch HW (1985) Biosynthesis and in vitro translation of the major surfactant-associated protein from human lung. J Biol Chem 260:495–500Google Scholar
  9. Goerke J (1974) Lung surfactant. Biochim Biophys Acta 344:241–261Google Scholar
  10. Graham RC Jr, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302Google Scholar
  11. Jaskoll TF, Phelps DS, Taeusch HW, Smith BT, Slavkin HC (1984) Localization of pulmonary surfactant protein during mouse lung development. Dev Biol 106:256–261Google Scholar
  12. Katyal SL, Singh G (1979) An immunologic study of the apoproteins of rat lung surfactant. Lab Invest 40:562–567Google Scholar
  13. Katyal SL, Singh G (1981) Analysis of pulmonary surfactant apoproteins by electrophoresis. Biochim Biophys Acta 670:323–331Google Scholar
  14. Katyal SL, Singh G (1984) Structural and ontogenic relationships of rat lung surfactant apoproteins. Exp Lung Res 6:175–189Google Scholar
  15. Katyal SL, Singh G (1985) In vitro translation of rat lung surfactant apoprotein mRNA. Biochem Biophys Res Commun 127:106–111Google Scholar
  16. Kessler W (1975) Rapid isolation of antigens from cells with a Staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol 115:1617–1624Google Scholar
  17. King RJ (1974) The surfactant system of the lung. Fed. Proc 33:2238–2247Google Scholar
  18. King RJ (1984a) Isolation and chemical composition of pulmonary surfactant. In: Robertson B, Golde LMG van, Batenburg JJ (eds) Pulmonary surfactant. Elsevier, Amsterdam, pp 1–15Google Scholar
  19. King RJ (1984b) The apolipoproteins of pulmonary surfactant. In: Wichert P von (ed) Current concepts in surfactant research. Prog Respir Res 18:68–82Google Scholar
  20. King RJ, Martin H (1980) Intracellular metabolism of the apoproteins of pulmonary surfactant in rat lung. J Appl Physiol 48:812–820Google Scholar
  21. King RJ, Klass DJ, Gikas EG, Clements JA (1973) Isolation of apoproteins from canine surface active material. Am J Physiol 224:788–795Google Scholar
  22. King RJ, Martin H, Mitts D, Holmstrom FM (1977) Metabolism of the apoproteins in pulmonary surfactant. J Appl Physiol 42:483–491Google Scholar
  23. Klass DJ (1973) Immunochemical studies of the protein fraction of pulmonary surface active material. Am Rev Respir Dis 107:784–789Google Scholar
  24. Laemmli NK, Favre M (1973) Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol 80:575–590Google Scholar
  25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  26. Mason R, Jones S, Benson B, Gikas E, Williams M, Clements J (1978) Immunofluorescent localization of purified surface active material in rat lung and isolated type II cells. Clin Res 26:451a (Abstract)Google Scholar
  27. Nierhaus KH (1982) Structure, assembly and function of ribosomes. In: Henke W, Hofschneider PH, Koprowski H, Melchers F, Rott R, Schweiger HG, Vogt PK (eds) Current topics in microbiology and immunology, Vol 97. Springer, Berlin Heidelberg New York, pp 80–155Google Scholar
  28. Phelps DS, Taeusch HW Jr, Benson B, Hawgood S (1984) An electrophoretic and immunochemical characterization of human surfactant-associated proteins. Biochim Biophys Acta 791:226–238Google Scholar
  29. Phizackerley PJR, Town MH, Newman GE (1979) Hydrophobic proteins of lamellated osmiophilic bodies isolated from pig lung. Biochem J 183:731–736Google Scholar
  30. Roobol K, Möller W (1978) Transient interaction between elongation factor 1 from Artemia salina and the 80 S ribosome. FEBS Lett 96:377–380Google Scholar
  31. Rooney SA (1985) The surfactant system and lung phospholipid biochemistry. Am Rev Respir Dis 131:439–460Google Scholar
  32. Shelley SA, Balis JU, Paciga JE, Espinoza CG, Richman AV (1982) Biochemical composition of adult human lung surfactant. Lung 160:195–206Google Scholar
  33. Slobin LI (1980) The role of eucaryotic elongation factor Tu in protein synthesis. Eur J Biochem 110:555–563Google Scholar
  34. Sueishi K, Tanaka K, Oda T (1977) Immunoultrastructural study of surfactant system. Distribution of specific protein of surface active material in rabbit lung. Lab Invest 37:136–142Google Scholar
  35. Sueishi K, Benson BJ (1981) Isolation of a major apolipoprotein of canine and murine pulmonary surfactant: biochemical and immunochemical characteristics. Biochim Biophys Acta 665:442–453Google Scholar
  36. Ten Have-Opbroek AAW (1975) Immunological study of lung development in the mouse embryo. I. Appearance of a lung-specific antigen, localized in the great alveolar cell. Dev Biol 46:390–403Google Scholar
  37. Ten Have-Opbroek AAW (1979) Immunological study of lung development in the mouse embryo. II. First appearance of the great alveolar cell, as shown by immunofluorescence microscopy. Dev Biol 69:408–423Google Scholar
  38. Ten Have-Opbroek AAW (1981) The development of the lung in mammals. An analysis of concepts and findings. Am J Anat 162:201–219Google Scholar
  39. Ten Have-Opbroek AAW (1986) The structural composition of the pulmonary acinus in the mouse. A scanning electron microscopical and developmental-biological analysis. Anat Embryol 174:49–57Google Scholar
  40. Ten Have-Opbroek AAW, Van Hemert FJ, De Wolf GJ (1984) Specific antigenic marker and surfactant synthesis in the great alveolar cell. Anat Rec 208:180A (Abstract)Google Scholar
  41. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354Google Scholar
  42. Van Hemert FJ, Roobol K, Möller W (1981) Partial purification of the messenger RNA for eukaryotic elongation factor Tu from Artemia salian. Eur J Biochem 120:137–142Google Scholar
  43. Van Hemert FJ, Amons RM, Pluijms WJM, Van Ormondt H, Möller W (1984) The primary structure of elongation factor EF-1α from the brine shrimp Artemia. EMBO J 3:1109–1113Google Scholar
  44. Weaver TE, Whitsett JA, Hull WM, Ross G (1985) Identification of canine pulmonary surfactant-associated glycoprotein A precursors. J Appl Physiol 58:2091–2095Google Scholar
  45. Whitsett JA, Weaver T, Hull W, Ross G, Dion C (1985a) Synthesis of surfactant-associated glycoprotein A by rat type II epithelial cells. Primary translation products and post-translational modification. Biochim Biophys Acta 828:162–171Google Scholar
  46. Whitsett JA, Ross G, Weaver T, Rice W, Dion C, Hull W (1985b) Glycosylation and secretion of surfactant-associated glycoprotein A. J Biol Chem 260:15273–15279Google Scholar
  47. Williams MC, Benson BJ (1981) Immunocytochemical localization and identification of the major surfactant protein in adult rat lung. J Histochem Cytochem 29:291–305Google Scholar
  48. Wright JR, Benson BJ, Williams MC, Goerke J, Clements JA (1984) Protein composition of rabbit alveolar surfactant subfractions. Biochim Biophys Acta 791:320–332Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • F. J. Van Hemert
    • 2
  • A. A. W. Ten Have-Opbroek
    • 1
  • C. J. M. Otto-Verberne
    • 1
  1. 1.Department of Anatomy and EmbryologyUniversity of LeidenLeidenThe Netherlands
  2. 2.Department of Physiological ChemistryUniversity of LeidenLeidenThe Netherlands

Personalised recommendations