Advertisement

Histochemistry

, Volume 97, Issue 3, pp 283–292 | Cite as

Synthesis, secretion and immunoelectron microscopic demonstration of apolipoprotein B-containing lipoprotein particles in the visceral rat yolk sac

  • H. Franke
  • D. Plonné
  • L. Winkler
  • R. Dargel
Article

Summary

Electron microscopic investigations on the involvement of the fetal membranes of the rat (visceral yolk sac) in the lipid metabolism revealed the occurrence of lipoprotein-sized particles located in cisternal Golgi stacks, Golgi vesicles and secretory vesicles of the cells of the visceral yolk sac epithelium as well as in distended areas of the intercellular space between adjacent epithelial cells. Application of the protein A-gold technique with specific anti-apoB antiserum resulted in a specific location of immunogold both over the different compartments of the lipoprotein pathway (RER, Golgi complex, secretory vesicles) as well as over the distended intercellular spaces, thus confirming these particles to be lipoproteins in nature. Isolated visceral epithelial cells prepared by a tryptic digestion method exhibited some ultrastructural alterations, such as a loss of apical brush border, a change from columnar to spherical cell shape, a decrease in phagolysosomes, but an increase in autophagosomal structures after 6 h incubation at a vitality rate of at least 85%. Within this period the epithelial cells secreted measurable amounts of apoB-containing lipoproteins into the medium floating in the density classes d<1.006 g/ml, d=1.006–1.020 g/ml and d=1.020–1.064 g/ml. The production of the lipoproteins was partly inhibited by cycloheximide indicating the secretion of particles with preformed as well as newly synthesized apoB. Negative staining of the particles revealed an average diameter of 34 nm of VLDL, 31 nm of IDL and 24 nm of LDL. In summary, our studies demonstrate that in the feto-placental unit of the rat the fetal membranes are capable of synthesizing and secreting lipoproteins. The cells of the visceral yolk sac epithelium were shown to be the producers of apoB-containing particles.

Keywords

Secretory Vesicle Fetal Membrane Golgi Vesicle Golgi Stack Microscopic Demonstration 
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.

Abbreviations

apo

apolipoprotein

ER

endoplasmic reticulum

IDL

intermediate density-lipoprotein

LDL

low density-lipoprotein

VLDL

very low density-lipoprotein

PBS

phosphate-buffered salt solution

RER

rough endoplasmic reticulum

TEM

transmission electron microscopy

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Angermüller S, Fahimi HD (1982) Imidazole-buffered osmium tetroxide: an excellent stain for visualization of lipids in TEM. Histochem J 14:823–835Google Scholar
  2. Belknap WM, Dietschy JM (1988) Sterol synthesis and LDL-clearance in vivo in the pregnant rat, placenta and fetus. J Clin Invest 51:1528–1536Google Scholar
  3. Bendayan M (1980) Use of the protein A-gold technique for the morphological study of vascular permeability. J Histochem Cytochem 28:1252–1254Google Scholar
  4. Bendayan M, Zollinger M (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J Histochem Cytochem 31:101–111Google Scholar
  5. Chao YS, Windler EE, Chen GC, Havel JR (1979) Hepatic catabolism of rat and human lipoproteins in rats treated with 17-α ethinyl estradiol. J Biol Chem 254:11360–11366Google Scholar
  6. Chevallier F (1964) Transferts et synthese du cholesterol chez le rat au cours de sa croissance. Biochim Biophys Acta 84:316–339Google Scholar
  7. Dargel R, Plonné D, Schlag B, Franke H, Winkler L (1988) Rat LDL metabolism in the perinatal period. In: Parnham MJ, Nieman R (eds) Cologne atherosclerosis conference, no. 4. Cholesterol homeostasis. Birkhäuser, Basel, pp 109–115Google Scholar
  8. Demmer LA, Levin MS, Elovson J, Reuben MA, Lusis AJ, Gordon JI (1986) Tissue-specific expression and developmental regulation of the rat apolipoprotein B gene. Proc Natl Acad Sci USA 83:8102–8106Google Scholar
  9. Dürer U, Franke H, Dargel R, Ude J (1986) A post-embedding immunoelectron-microscopic demonstration of apolipoprotein-B-containing lipoprotein particles in hepatocytes of fetal rats. Histochem 84:263–370Google Scholar
  10. Dunton A, Al-Alousi LA, Pratten MK, Beck F (1986) The giant yolk sac: a model for studying early placental transport. J Anat 145:189–206Google Scholar
  11. Elshourbagy NA, Boguski NA, Liao WSL, Jefferson LD, Gordon JL, Taylor JM (1985) Expression of rat apolipoprotein A IV and AI genes: mRNA induction during development and in response to glucocorticoids and insulin. Proc Natl Acad Sci USA 83:8242–8246Google Scholar
  12. Finck W, Theil S (1964) Die embryonale Erythropoese und der Nukleinsäuregehalt in der embryonalen Leber bei ganzkörperbestrahlten Ratten. Acta Biol Med Germ 12:354–364Google Scholar
  13. Franke H (1979) Die Wirkung einer Unterbrechung der Dottersackzirkulation auf die Ultrastruktur des Dottersackes und die Entwicklung des Feten bei der Ratte. Acta Anat 103:200–211Google Scholar
  14. Franke H, Dargel R (1985) Morphometric studies on lipoprotein particles in developing rat liver and their corticosteroid-induced changes during the late gestational period. Cell Tissue Res 242:661–667Google Scholar
  15. Franke H, Goetze E (1963) Die Feinstruktur der Leberzellen von Rattenfoeten und Neugeborenen in verschiedenen Entwicklungsstadien. Acta Biol Med Germ 11:424–432Google Scholar
  16. Franke H, Oswald B (1973) Electron microscopic studies on the fine structure of the visceral yolk sac epithelium of the rat following an intrauterine injection of a rabbit antirat placental serum or a normal serum. Beitr Pathol 148:1–14Google Scholar
  17. Franke H, Goetze T, Goetze E (1975) Fine structure and pinocytic activity of the visceral epithelium of the rat yolk sac incubated in a culture medium with or without goat antirat-placenta-IgG. Beitr Pathol 154:170–181Google Scholar
  18. Franke H, Tönjes R, Goetze T, Goetze E (1976) Evidence for the occurrence of microtubules in the visceral yolk sac epithelium. Cytobiology 12:476–480Google Scholar
  19. Franke H, Poli G, Zimmermann T, Dianzani MU, Dargel R (1988) Short-term effects of carbon tetrachloride on the lipoprotein secretion in isolated rat hepatocytes. Virchows Arch [B] 54:357–365Google Scholar
  20. Franke H, Müller D, Schlag B, Zimmermann T, Dargel R (1990) Biochemical and ultrastructural studies on the effect of verapamil on formation and secretion of lipoproteins in rat hepatocyte suspension. Arch Toxicol 64:656–662Google Scholar
  21. Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature 241:20–22Google Scholar
  22. Goetze T, Franke H, Hühn W, Tönjes R (1976) Density gradient separation, submicroscopic and enzymatic characterization of cell organelles involved in the protein uptake and degradation in the rat yolk sac after culturing with 125I-labeled human serum albumin. Cytobiology 12:274–286Google Scholar
  23. Hornick CA, Hamilton RL, Spaziani E, Enders GH, Havel RJ (1985) Isolation and characterization of multivesicular bodies from rat hepatocytes: an organelle distinct from secretory vesicles of the Golgi apparatus. J Cell Biol 100:1558–1569Google Scholar
  24. Hühn W, Franke H, Goetze T, Tönjes R (1975) Die Auftrennung der Zellorganellen der Eihaut (viszeraler Dottersack) der Ratte. Acta Histochem Suppl (Jena) 15:41–48Google Scholar
  25. Jones AL, Ockner RK (1971) An electron microscopic study of endogenous very low density lipoprotein production in the intestine of rat and man. J Lipid Res 12:580–589Google Scholar
  26. Lanford RE, Bronson DL, Estlack LE, Wians FH Jr (1991) Plasma protein and apolipoprotein synthesis by human yolk sac carcinoma cells in vitro. In Vitro Cell Dev Biol 27A:205–210Google Scholar
  27. Luft JH (1961) Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol 9:409Google Scholar
  28. Mak KM, Trier JS (1975) Lipoprotein particles in the jejunal mucosa of fetal rats. Develop Biol 43:204–211Google Scholar
  29. Mann HB, Whitney DR (1947) On a test whether one of two random variables is stochastically larger than the other. Ann Math Stat 18:52–54Google Scholar
  30. Ouchterlony O (1949) Antigen-antibody reactions in gels. Acta Pathol Microbiol Scand 26:507–516Google Scholar
  31. Palade HA, Deren JJ, Wilson TH (1966) I. Development of structure and function in the mammalian yolk sac. I. Developmental morphology and vitamin B12 uptake of the rat yolk sac. Dev Biol 3:311–348Google Scholar
  32. Pelletier G, Puviani R, Bosler O, Descarries L (1981) Immunocytochemical detection of peptides in osmicated and plastic-embedded tissue. An electron microscopic study. J Histochem Cytochem 29:759–771Google Scholar
  33. Petry G, Zuleger S (1958) Vergleichende Studie über den Bau der Eihüllen der Ratte. Z Zellforsch 47:683–712Google Scholar
  34. Plonné D, Schlag B, Winkler L, Dargel R (1990) Tracer kinetic studies of the low density lipoprotein metabolism in the fetal rat: an example for estimation of flux rates in the nonsteady state. J Lipid Res 31:747–752Google Scholar
  35. Schlag B, Winkler L (1973) Low density lipoprotein of the rat fetus. Acta Biol Med Germ 30:477–482Google Scholar
  36. Schlag B, Winkler L (1978) Konzentration und Zusammensetzung der Lipoproteinklassen der fetalen Ratte. Acta Biol Med Germ 37:233–237Google Scholar
  37. Schlag B, Winkler L, Plonné D, Dürer U, Dargel R (1987a) Preparation of apo E-free rat low density lipoprotein for catabolic studies. J Lipid Res 28:1521–1524Google Scholar
  38. Schlag B, Winkler L, Dargel R (1987b) Very low density lipoproteins in the fetal rat at term. Biomed Biochim Acta 46:83–87Google Scholar
  39. Shi WK, Heath JK (1984) Apolipoprotein expression by murine visceral yolk sac endoderm. J Embryol Exp Morphol 81:143–152Google Scholar
  40. Zimmermann T, Franke H, Dargel R (1987) Isolation and characterization of parenchymal cells from normal and cirrhotic rat liver. Cell Biochem Funct 5:47–54Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • H. Franke
    • 1
  • D. Plonné
    • 1
  • L. Winkler
    • 1
  • R. Dargel
    • 1
  1. 1.Institute of Pathological BiochemistryFriedrich-Schiller UniversityJenaFederal Republic of Germany

Personalised recommendations