Abstract
Chylomicrons containing labeled cholesterol, mainly (70%) present as cholesteryl ester, were injected intravenously into intact rats, and samples of liver were obtained 27–210 min later. Most (58–75%) of the injected label was recovered in the liver after 27–75 min. Hepatic uptake occurred without hydrolysis of the labeled cholesteryl ester. In separate experiments, in vitro perfusion of livers of similarly treated rats for 30–35 min washed out only 3–9% of the labeled sterol. Samples of liver and small intestine were prepared for electron microscopy with Aquon as the dehydrating agent. Good retention (70% or more) of labeled cholesterol and satisfactory preservation of ultrastructure were obtained. After 30 min, the radioautographic reaction was localized mainly over the region of the cell boundary of the parenchymal liver cells, with fewer grains being present over intracellular organelles. At later time intervals, when considerable hydrolysis of the labeled cholesteryl ester had occurred, the radioautographic reaction was more evenly distributed. Phagocytosed labeled lipid was seen in Kupffer cells after the larger lipid load; phagocytosis by parenchymal cells was not seen. In other experiments, cholesteryl ester hydrolase activity was found in all subcellular fractions, the microsome and plasma membrane fractions showing the highest activity per mg protein. The mechanism of cholesteryl ester transport into the liver cell may involve: (1) hydrolysis at the cell surface; or (2) slow entry of intact molecules followed by intracellular hydrolysis of the ester bond.
Full Text
The Full Text of this article is available as a PDF (3.2 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BELFRAGE P., BORGSTROM B., OLIVERCRONA T. The tissue distribution of radioactivity following the injection of varying levels of fatty acid labeled chylomicrons in the rat. Acta Physiol Scand. 1963 Jun-Jul;58:111–123. doi: 10.1111/j.1748-1716.1963.tb02634.x. [DOI] [PubMed] [Google Scholar]
- BENACERRAF B., BIOZZI G., HALPERN B. N., NEVEU T., STIFFEL C. Role of reticulo-endothelial system in blood clearance of cholesterol. Am J Physiol. 1956 Nov;187(2):269–274. doi: 10.1152/ajplegacy.1956.187.2.269. [DOI] [PubMed] [Google Scholar]
- 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]
- BORGSTROM B., JORDAN P. Metabolism of chylomicron glyceride as studied by C14-glycerol-C14-palmitic acid labeled chylomicrons. Acta Soc Med Ups. 1959;64:185–193. [PubMed] [Google Scholar]
- Belfrage P. Metabolism of chyle triglycerides in the liver. I. Studies on the mechanisms for liver uptake of intravenously injected, glycerol- and fatty acid-labeled chyle in the carbohydrate-fed rat. Biochim Biophys Acta. 1966 Dec 7;125(3):474–484. [PubMed] [Google Scholar]
- CARO L. G., VAN TUBERGEN R. P., KOLB J. A. High-resolution autoradiography. I. Methods. J Cell Biol. 1962 Nov;15:173–188. doi: 10.1083/jcb.15.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CAULFIELD J. B. Effects of varying the vehicle for OsO4 in tissue fixation. J Biophys Biochem Cytol. 1957 Sep 25;3(5):827–830. doi: 10.1083/jcb.3.5.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
- DEYKIN D., GOODMAN D. S. The hydrolysis of long-chain fatty acid esters of cholesterol with rat liver enzymes. J Biol Chem. 1962 Dec;237:3649–3656. [PubMed] [Google Scholar]
- Elovson J., Olivecrona T., Belfrage P. Metabolism in the rat of chyle obtained after feeding hydrogenated coconut oil labeled with stearic acid. Biochim Biophys Acta. 1965 Jul 7;106(1):34–44. doi: 10.1016/0005-2760(65)90093-7. [DOI] [PubMed] [Google Scholar]
- Forstner G. G., Sabesin S. M., Isselbacher K. J. Rat intestinal microvillus membranes. Purification and biochemical characterization. Biochem J. 1968 Jan;106(2):381–390. doi: 10.1042/bj1060381. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GIBBONS I. R. An embedding resin miscible with water for electron microscopy. Nature. 1959 Aug 1;184(Suppl 6):375–376. doi: 10.1038/184375b0. [DOI] [PubMed] [Google Scholar]
- GOODMAN D. S. The metabolism of chylomicron cholesterol ester in the rat. J Clin Invest. 1962 Oct;41:1886–1896. doi: 10.1172/JCI104645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GORNALL A. G., BARDAWILL C. J., DAVID M. M. Determination of serum proteins by means of the biuret reaction. J Biol Chem. 1949 Feb;177(2):751–766. [PubMed] [Google Scholar]
- GREEN C., WEBB J. A. THE UPTAKE OF CHYLOMICRON FATTY ACIDS BY ISOLATED LIVER CELLS. Biochim Biophys Acta. 1964 Aug 5;84:404–411. doi: 10.1016/0926-6542(64)90004-6. [DOI] [PubMed] [Google Scholar]
- Higgins J. A. Forces involved in chylomicron binding by isolated cells of rat liver. J Lipid Res. 1967 Nov;8(6):636–641. [PubMed] [Google Scholar]
- Higgins J. A., Green C. Properties of a lipase of rat-liver parenchymal cells. Biochim Biophys Acta. 1967 Oct 2;144(2):211–220. doi: 10.1016/0005-2760(67)90151-8. [DOI] [PubMed] [Google Scholar]
- Higgins J. A., Green C. The uptake of lipids by rat liver cells. Biochem J. 1966 Jun;99(3):631–639. doi: 10.1042/bj0990631. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- MILLER L. L., BLY C. G., WATSON M. L., BALE W. F. The dominant role of the liver in plasma protein synthesis; a direct study of the isolated perfused rat liver with the aid of lysine-epsilon-C14. J Exp Med. 1951 Nov;94(5):431–453. doi: 10.1084/jem.94.5.431. [DOI] [PMC free article] [PubMed] [Google Scholar]
- NESTEL P. J., HAVEL R. J., BEZMAN A. METABOLISM OF CONSTITUENT LIPIDS OF DOG CHYLOMICRONS. J Clin Invest. 1963 Aug;42:1313–1321. doi: 10.1172/JCI104815. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nestel P. J., Havel R. J., Bezman A. SITES OF INITIAL REMOVAL OF CHYLOMICRON TRIGLYCERIDE FATTY ACIDS FROM THE BLOOD. J Clin Invest. 1962 Oct;41(10):1915–1921. doi: 10.1172/JCI104648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- OLIVECRONA T., BELFRAGE P. MECHANISMS FOR REMOVAL OF CHYLE TRIGLYCERIDE FROM THE CIRCULATING BLOOD AS STUDIED WITH (14C)GLYCEROL AND (3H)PALMITIC ACID- LABELED CHYLE. Biochim Biophys Acta. 1965 Feb 1;98:81–93. doi: 10.1016/0005-2760(65)90013-5. [DOI] [PubMed] [Google Scholar]
- Ontko J. A. Chylomicron, free fatty acid and ketone body metabolism of isolated liver cells and liver homogenates. Biochim Biophys Acta. 1967 Feb 14;137(1):13–22. doi: 10.1016/0005-2760(67)90003-3. [DOI] [PubMed] [Google Scholar]
- Ontko J. A., Zilversmit D. B. Metabolism of chylomicrons by the isolated rat liver. J Lipid Res. 1967 Mar;8(2):90–96. [PubMed] [Google Scholar]
- Quarfordt S. H., Goodman D. S. Heterogeneity in the rate of plasma clearance of chylomicrons of different size. Biochim Biophys Acta. 1966 Apr 4;116(2):382–385. doi: 10.1016/0005-2760(66)90019-1. [DOI] [PubMed] [Google Scholar]
- Quarfordt S. H., Goodman D. S. Metabolism of doubly-labeled chylomicron cholesteryl esters in the rat. J Lipid Res. 1967 May;8(3):264–273. [PubMed] [Google Scholar]
- REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
- STEIN Y., SHAPIRO B. Uptake and metabolism of triglycerides by the rat liver. J Lipid Res. 1960 Jul;1:326–331. [PubMed] [Google Scholar]
- STERN I., SHAPIRO B. A rapid and simple method for the determination of esterified fatty acids and for total fatty acids in blood. J Clin Pathol. 1953 May;6(2):158–160. doi: 10.1136/jcp.6.2.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schotz M. C., Arnesjö B., Olivecrona T. The role of the liver in the uptake of plasma and chyle triglycerides in the rat. Biochim Biophys Acta. 1966 Dec 7;125(3):485–495. doi: 10.1016/0005-2760(66)90037-3. [DOI] [PubMed] [Google Scholar]
- Stein O., Stein Y. Lipid synthesis, intracellular transport, storage, and secretion. I. Electron microscopic radioautographic study of liver after injection of tritiated palmitate or glycerol in fasted and ethanol-treated rats. J Cell Biol. 1967 May;33(2):319–339. doi: 10.1083/jcb.33.2.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stein O., Stein Y. The role of the liver in the metabolism of chylomicrons, studied by electron microscopic autoradiography. Lab Invest. 1967 Oct;17(4):436–446. [PubMed] [Google Scholar]
- Stein Y., Widnell C., Stein O. Acylation of lysophosphatides by plasma membrane fractions of rat liver. J Cell Biol. 1968 Oct;39(1):185–192. doi: 10.1083/jcb.39.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]