Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1971 Oct 1;51(1):1–25. doi: 10.1083/jcb.51.1.1

SITES OF LIPOPROTEIN LIPASE ACTIVITY IN ADIPOSE TISSUE PERFUSED WITH CHYLOMICRONS

Electron Microscope Cytochemical Study

E Joan Blanchette-Mackie 1, Robert O Scow 1
PMCID: PMC2108247  PMID: 4329521

Abstract

Lipoprotein lipase activity was studied in rat parametrial adipose tissue perfused with chylomicrons and in gelatin blocks containing postheparin plasma and chylomicrons. The tissues and blocks were fixed in glutaraldehyde and incubated in 0.035 M CaCl2-0.1 M Tris medium (pH 8.3) at 38°C. The doubly labeled chylomicron triglycerides (glycerol-3H and palmitate-14C) in the tissues and blocks were hydrolyzed during incubation to free fatty acids (FFA) and the FFA remained in the specimens; hydrolysis was inhibited by 0.004 M diethyl paranitrophenyl phosphate (E-600). Incubated blocks and tissue were treated with 0.05 M Pb(NO3)2, postfixed in OsO4, dehydrated with acetone, embedded in Epon, and examined by electron microscopy. The incubated blocks contained electronlucent areas and granular and laminar precipitates at sites of hydrolysis. Similar precipitates were found in incubated tissue, within vacuoles and microvesicles of capillary endothelium, and in the subendothelial space (between the endothelium and pericytes), but not in the capillary lumen or in or near fat cells. The cytochemical reaction was greatly reduced, in blocks and tissues incubated with E-600. It is concluded that plasma glycerides are hydrolyzed by lipoprotein lipase in capillary endothelial cells and in the subendothelial space of adipose tissue and that glycerides across the endothelial cells within a membrane-bounded system.

Full Text

The Full Text of this article is available as a PDF (2.9 MB).

Selected References

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

  1. BARRY J. M., BARTLEY W., LINZELL J. L., ROBINSON D. S. THE UPTAKE FROM THE BLOOD OF TRIGLYCERIDE FATTY ACIDS OF CHYLOMICRA AND LOW-DENSITY LIPOPROTEINS BY THE MAMMARY GLAND OF THE GOAT. Biochem J. 1963 Oct;89:6–11. doi: 10.1042/bj0890006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BORGSTROM B. Investigation on lipid separation methods. Separation of cholesterol esters, glycerides and free fatty acids. Acta Physiol Scand. 1952 Jun 6;25(2-3):111–119. doi: 10.1111/j.1748-1716.1952.tb00863.x. [DOI] [PubMed] [Google Scholar]
  3. Chernick S., Novak M. Effect of insulin on FFA mobilization and ketosis in fasting pregnant rats. Diabetes. 1970 Aug;19(8):563–570. doi: 10.2337/diab.19.8.563. [DOI] [PubMed] [Google Scholar]
  4. Cunningham V. J., Robinson D. S. Clearing-factor lipase in adipose tissue. Distinction of different states of the enzyme and the possible role of the fat cell in the maintenance of tissue activity. Biochem J. 1969 Apr;112(2):203–209. doi: 10.1042/bj1120203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DOLE V. P., MEINERTZ H. Microdetermination of long-chain fatty acids in plasma and tissues. J Biol Chem. 1960 Sep;235:2595–2599. [PubMed] [Google Scholar]
  6. Enser M. B., Kunz F., Borensztajn J., Opie L. H., Robinson D. S. Metabolism of triglyceride fatty acid by the perfused rat heart. Biochem J. 1967 Jul;104(1):306–317. doi: 10.1042/bj1040306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  8. FREDRICKSON D. S., GORDON R. S., Jr Transport of fatty acids. Physiol Rev. 1958 Oct;38(4):585–630. doi: 10.1152/physrev.1958.38.4.585. [DOI] [PubMed] [Google Scholar]
  9. Fain J. N., Reed N., Saperstein R. The isolation and metabolism of brown fat cells. J Biol Chem. 1967 Apr 25;242(8):1887–1894. [PubMed] [Google Scholar]
  10. Fredrickson D. S., Levy R. I., Lees R. S. Fat transport in lipoproteins--an integrated approach to mechanisms and disorders. N Engl J Med. 1967 Jan 19;276(3):148–contd. doi: 10.1056/NEJM196701192760305. [DOI] [PubMed] [Google Scholar]
  11. HOLLENBERG C. H. Effect of nutrition on activity and release of lipase from rat adipose tissue. Am J Physiol. 1959 Sep;197:667–670. doi: 10.1152/ajplegacy.1959.197.3.667. [DOI] [PubMed] [Google Scholar]
  12. Hamosh M., Clary T. R., Chernick S. S., Scow R. O. Lipoprotein lipase activity of adipose and mammary tissue and plasma triglyceride in pregnant and lactating rats. Biochim Biophys Acta. 1970 Sep 8;210(3):473–482. doi: 10.1016/0005-2760(70)90044-5. [DOI] [PubMed] [Google Scholar]
  13. Ho S. J., Ho R. J., Meng H. C. Comparison of heparin-released and epinephrine-sensitive lipases in rat adipose tissue. Am J Physiol. 1967 Feb;212(2):284–290. doi: 10.1152/ajplegacy.1967.212.2.284. [DOI] [PubMed] [Google Scholar]
  14. KESSLER J. I. Effect of diabetes and insulin on the activity of myocardial and adipose tissue lipoprotein lipase of rats. J Clin Invest. 1963 Mar;42:362–367. doi: 10.1172/JCI104722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. KORN E. D., QUIGLEY T. W., Jr Lipoprotein lipase of chicken adipose tissue. J Biol Chem. 1957 Jun;226(2):833–839. [PubMed] [Google Scholar]
  16. LUCY J. A., GLAUERT A. M. STRUCTURE AND ASSEMBLY OF MACROMOLECULAR LIPID COMPLEXES COMPOSED OF GLOBULAR MICELLES. J Mol Biol. 1964 May;8:727–748. doi: 10.1016/s0022-2836(64)80121-2. [DOI] [PubMed] [Google Scholar]
  17. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McBride O. W., Korn E. D. The uptake of doubly labeled chylomicrons by guinea pig mammary gland and liver. J Lipid Res. 1964 Jul;5(3):459–467. [PubMed] [Google Scholar]
  19. Nakajima O., Gray C. H. Studies on haem alpha-methenyl oxygenase. Isomeric structure of formylbiliverdin, a possible precursor of biliverdin. Biochem J. 1967 Jul;104(1):20–22. doi: 10.1042/bj1040020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pokrajac N., Lossow W. J., Chaikoff I. L. The effect of nutritional state on lipoprotein lipase activity in isolated rat adipose tissue cells. Biochim Biophys Acta. 1967 May 16;139(1):123–132. doi: 10.1016/0005-2744(67)90118-0. [DOI] [PubMed] [Google Scholar]
  21. ROBERT A., SCOW R. O. PERFUSION OF RAT ADIPOSE TISSUE. Am J Physiol. 1963 Aug;205:405–412. doi: 10.1152/ajplegacy.1963.205.2.405. [DOI] [PubMed] [Google Scholar]
  22. ROBINSON D. S. THE CLEARING FACTOR LIPASE AND ITS ACTION IN THE TRANSPORT OF FATTY ACIDS BETWEEN THE BLOOD AND TISSUES. Adv Lipid Res. 1963;1:133–182. doi: 10.1016/b978-1-4831-9937-5.50010-7. [DOI] [PubMed] [Google Scholar]
  23. RODBELL M. LOCALIZATION OF LIPOPROTEIN LIPASE IN FAT CELLS OF RAT ADIPOSE TISSUE. J Biol Chem. 1964 Mar;239:753–755. [PubMed] [Google Scholar]
  24. RODBELL M., SCOW R. O. METABOLISM OF CHYLOMICRONS AND TRIGLYCERIDE EMULSIONS BY PERFUSED RAT ADIPOSE TISSUE. Am J Physiol. 1965 Jan;208:106–114. doi: 10.1152/ajplegacy.1965.208.1.106. [DOI] [PubMed] [Google Scholar]
  25. SCHNATZ J. D., WILLIAMS R. H. The effect of acute insulin deficiency in the rat on adipose tissue lipolytic activity and plasma lipids. Diabetes. 1963 Mar-Apr;12:174–178. doi: 10.2337/diab.12.2.174. [DOI] [PubMed] [Google Scholar]
  26. Salaman M. R., Robinson D. S. Clearing-factor lipase in adipose tissue. A medium in which the enzyme activity of tissue from starved rats increases in vitro. Biochem J. 1966 Jun;99(3):640–647. doi: 10.1042/bj0990640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Scow R. O., Stein Y., Stein O. Incorporation of dietary lecithin and lysolecithin into lymph chylomicrons in the rat. J Biol Chem. 1967 Nov 10;242(21):4919–4924. [PubMed] [Google Scholar]
  28. Stein O., Scow R. O., Stein Y. FFA-3H uptake by perfused adipose tissue: electron microscopic autoradiographic study. Am J Physiol. 1970 Aug;219(2):510–518. doi: 10.1152/ajplegacy.1970.219.2.510. [DOI] [PubMed] [Google Scholar]
  29. VENABLE J. H., COGGESHALL R. A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY. J Cell Biol. 1965 May;25:407–408. doi: 10.1083/jcb.25.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. WASSERMANN F., McDONALD T. F. Electron microscopic study of adipose tissue (fat organs) with special reference to the transport of lipids between blood and fat cells. Z Zellforsch Mikrosk Anat. 1963;59:326–357. doi: 10.1007/BF00339791. [DOI] [PubMed] [Google Scholar]
  31. WILLIAMSON J. R. ADIPOSE TISSUE. MORPHOLOGICAL CHANGES ASSOCIATED WITH LIPID MOBILIZATION. J Cell Biol. 1964 Jan;20:57–74. doi: 10.1083/jcb.20.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zilversmit D. B. The composition and structure of lymph chylomicrons in dog, rat, and man. J Clin Invest. 1965 Oct;44(10):1610–1622. doi: 10.1172/JCI105267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zilversmit D. B. The surface coat of chylomicrons: lipid chemistry. J Lipid Res. 1968 Mar;9(2):180–186. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES