Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1972 Jan 1;52(1):15–40. doi: 10.1083/jcb.52.1.15

EFFECTS OF PUROMYCIN ON THE STRUCTURE OF RAT INTESTINAL EPITHELIAL CELLS DURING FAT ABSORPTION

Harold I Friedman 1, Robert R Cardell Jr 1
PMCID: PMC2108684  PMID: 4331298

Abstract

This report provides information on the morphology of rat intestinal epithelial cells during fat absorption. In addition, the role of protein metabolism in this process has been evaluated by blocking its synthesis with puromycin and studying the fine structure of mucosal cells from rats at various times after fat intubation. The results indicate that SER-derived vesicles, containing fat droplets, migrate from the apical cytoplasm of the absorptive cell and fuse with saccules or vacuoles of the Golgi complex. Arguments are made that the Golgi complex is important in completing chylomicron formation and in providing appropriate enveloping membranes for the chylomicron. Such membranes may be necessary for Golgi vacuoles to fuse with the lateral cell membranes and release chylomicra. Puromycin treatment causes the absorptive cell to accumulate increased quantities of lipid that are devoid of membrane during fat absorption. In addition, puromycin-treated cells contain much less RER and Golgi membranes are strikingly decreased in number. In this paper we discuss the consequences of these abnormalities and suggest that continued protein synthesis by the RER is required in order to generate Golgi membranes. If such membranes are absent the cell's ability to discarge chylomicra is impaired and lipid accumulates.

Full Text

The Full Text of this article is available as a PDF (3.3 MB).

Selected References

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

  1. Ashley C. A., Feder N. Glycol methacrylate in histopathology. A study of central necrosis of the liver using a water-miscible plastic as embedding medium. Arch Pathol. 1966 May;81(5):391–397. [PubMed] [Google Scholar]
  2. Ashley C. A., Peters T., Jr Electron microscopic radioautographic detection of sites of protein synthesis and migration in liver. J Cell Biol. 1969 Nov;43(2):237–249. doi: 10.1083/jcb.43.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BOLLMAN J. L., FLOCK E. V., CAIN J. C., GRINDLAY J. H. Lipids of lymph following feeding of fat: an experimental study. Am J Physiol. 1950 Oct;163(1):41–47. doi: 10.1152/ajplegacy.1950.163.1.41. [DOI] [PubMed] [Google Scholar]
  4. BORGSTROM B., DAHLQVIST A., LUNDH G., SJOVALL J. Studies of intestinal digestion and absorption in the human. J Clin Invest. 1957 Oct;36(10):1521–1536. doi: 10.1172/JCI103549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. BORGSTROM B., LAURELL C. B. Studies of lymph and lymph-proteins during absorption of fat and saline by rats. Acta Physiol Scand. 1953 Oct 6;29(2-3):264–280. doi: 10.1111/j.1748-1716.1953.tb01023.x. [DOI] [PubMed] [Google Scholar]
  6. BORGSTROM B. On the mechanism of pancreatic lipolysis of glycerides. Biochim Biophys Acta. 1954 Apr;13(4):491–504. doi: 10.1016/0006-3002(54)90365-0. [DOI] [PubMed] [Google Scholar]
  7. Berlin J. D. The localization of acid mucopolysaccharides in the golgi complex of intestinal goblet cells. J Cell Biol. 1967 Mar;32(3):760–766. doi: 10.1083/jcb.32.3.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Borgström B. The dimensions of the bile salt micelle. Measurements by gel filtration. Biochim Biophys Acta. 1965 Jul 7;106(1):171–183. doi: 10.1016/0005-2760(65)90105-0. [DOI] [PubMed] [Google Scholar]
  9. Cardell R. R., Jr, Badenhausen S., Porter K. R. Intestinal triglyceride absorption in the rat. An electron microscopical study. J Cell Biol. 1967 Jul;34(1):123–155. doi: 10.1083/jcb.34.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Claude A. Growth and differentiation of cytoplasmic membranes in the course of lipoprotein granule synthesis in the hepatic cell. I. Elaboration of elements of the Golgi complex. J Cell Biol. 1970 Dec;47(3):745–766. doi: 10.1083/jcb.47.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Flickinger C. J. The development of Golgi complexes and their dependence upon the nucleus inmebae. J Cell Biol. 1969 Nov;43(2):250–262. doi: 10.1083/jcb.43.2.250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grove S. N., Bracker C. E., Morré D. J. Cytomembrane differentiation in the endoplasmic reticulum-Golgi apparatus-vesicle complex. Science. 1968 Jul 12;161(3837):171–173. doi: 10.1126/science.161.3837.171. [DOI] [PubMed] [Google Scholar]
  13. JOHNSTON J. M., BORGSTROEM B. THE INTESTINAL ABSORPTION AND METABOLISM OF MICELLAR SOLUTIONS OF LIPIDS. Biochim Biophys Acta. 1964 Aug 5;84:412–423. doi: 10.1016/0926-6542(64)90005-8. [DOI] [PubMed] [Google Scholar]
  14. Jamieson J. D., Palade G. E. Intracellular transport of secretory proteins in the pancreatic exocrine cell. I. Role of the peripheral elements of the Golgi complex. J Cell Biol. 1967 Aug;34(2):577–596. doi: 10.1083/jcb.34.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jersild R. A., Jr A time sequence study of fat absorption in the rat jejunum. Am J Anat. 1966 Jan;118(1):135–162. doi: 10.1002/aja.1001180108. [DOI] [PubMed] [Google Scholar]
  16. Jones A. L., Ruderman N. B., Herrera M. G. Electron microscopic and biochemical study of lipoprotein synthesis in the isolated perfused rat liver. J Lipid Res. 1967 Sep;8(5):429–446. [PubMed] [Google Scholar]
  17. Kayden H. J., Medick M. The absorption and metabolism of short and long chain fatty acids in puromycin-treated rats. Biochim Biophys Acta. 1969 Jan 21;176(1):37–43. doi: 10.1016/0005-2760(69)90070-8. [DOI] [PubMed] [Google Scholar]
  18. Kessel R. G. Some observations on the ultrastructure of the oocyte of Thyone briareus with special reference to the relationship of the Golgi complex and endoplasmic reticulum in the formation of yolk. J Ultrastruct Res. 1966 Oct;16(3):305–319. doi: 10.1016/s0022-5320(66)80065-5. [DOI] [PubMed] [Google Scholar]
  19. LACY D., TAYLOR A. B. Fat absorption by epithelial cells of the smal intestine of the rat. Am J Anat. 1962 Mar;110:155–185. doi: 10.1002/aja.1001100205. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. MATTSON F. H., BECK L. W. The digestion in vitro of triglycerides by pancreatic lipase. J Biol Chem. 1955 May;214(1):115–125. [PubMed] [Google Scholar]
  22. MATTSON F. H., BENEDICT J. H., MARTIN J. B., BECK L. W. Intermediates formed during the digestion of triglycerides. J Nutr. 1952 Nov;48(3):335–344. doi: 10.1093/jn/48.3.335. [DOI] [PubMed] [Google Scholar]
  23. MEEK G. A., BRADBURY S. Localization of thiamine pyrophosphatase activity in the golgi apparatus of a mollusc, Helix aspersa. J Cell Biol. 1963 Jul;18:73–85. doi: 10.1083/jcb.18.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. MOLLENHAUER H. H., WHALEY W. G. An observation on the functioning of the Golgi apparatus. J Cell Biol. 1963 Apr;17:222–225. doi: 10.1083/jcb.17.1.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Marsh J. B., Fritz R. The carbohydrate components of rat serum lipoproteins. Proc Soc Exp Biol Med. 1970 Jan;133(1):9–10. doi: 10.3181/00379727-133-34395. [DOI] [PubMed] [Google Scholar]
  26. Neutra M., Leblond C. P. Radioautographic comparison of the uptake of galactose-H and glucose-H3 in the golgi region of various cells secreting glycoproteins or mucopolysaccharides. J Cell Biol. 1966 Jul;30(1):137–150. doi: 10.1083/jcb.30.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neutra M., Leblond C. P. Synthesis of the carbohydrate of mucus in the golgi complex as shown by electron microscope radioautography of goblet cells from rats injected with glucose-H3. J Cell Biol. 1966 Jul;30(1):119–136. doi: 10.1083/jcb.30.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nichols A. V. Functions and interrelationships of different classes of plasma lipoproteins. Proc Natl Acad Sci U S A. 1969 Nov;64(3):1128–1137. doi: 10.1073/pnas.64.3.1128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. PADYKULA H. A. Recent functional interpretations of intestinal morphology. Fed Proc. 1962 Nov-Dec;21:873–879. [PubMed] [Google Scholar]
  30. PALAY S. L., KARLIN L. J. An electron microscopic study of the intestinal villus. II. The pathway of fat absorption. J Biophys Biochem Cytol. 1959 May 25;5(3):373–384. doi: 10.1083/jcb.5.3.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. PARSONS D. F. A simple method for obtaining increased contrast in araldite sections by using postfixation staining of tissues with potassium permanganate. J Biophys Biochem Cytol. 1961 Nov;11:492–497. doi: 10.1083/jcb.11.2.492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Rao G. A., Johnston J. M. Purification and properties of triglyceride synthetase from the intestinal mucosa. Biochim Biophys Acta. 1966 Dec 7;125(3):465–473. doi: 10.1016/0005-2760(66)90035-x. [DOI] [PubMed] [Google Scholar]
  34. Redgrave T. G., Zilversmit D. B. Does puromycin block release of chylomicrons from the intestine? Am J Physiol. 1969 Aug;217(2):336–340. doi: 10.1152/ajplegacy.1969.217.2.336. [DOI] [PubMed] [Google Scholar]
  35. Rodgers J. B., Riley E. M., Drummey G. D., Isselbacher K. J. Lipid absorption in adrenalectomized rats: the role of altered enzyme activity in the intestinal mucosa. Gastroenterology. 1967 Oct;53(4):547–556. [PubMed] [Google Scholar]
  36. SABESIN S. M., ISSELBACHER K. J. PROTEIN SYNTHESIS INHIBITION: MECHANISM FOR THE PRODUCTION OF IMPAIRED FAT ABSORPTION. Science. 1965 Mar 5;147(3662):1149–1151. doi: 10.1126/science.147.3662.1149. [DOI] [PubMed] [Google Scholar]
  37. SENIOR J. R. INTESTINAL ABSORPTION OF FATS. J Lipid Res. 1964 Oct;5:495–521. [PubMed] [Google Scholar]
  38. SENIOR J. R., ISSELBACHER K. J. Direct esterification of monoglycerides with palmityl coenzyme A by intestinal epithelial subcellular fractions. J Biol Chem. 1962 May;237:1454–1459. [PubMed] [Google Scholar]
  39. SIMMONDS W. J. The effect of fluid, electrolyte and food intake on thoracic duct lymph flow in unanaesthetized rats. Aust J Exp Biol Med Sci. 1954 Jun;32(3):285–299. doi: 10.1038/icb.1954.32. [DOI] [PubMed] [Google Scholar]
  40. Sjöstrand F. S., Borgström B. The lipid components of the smooth-surfaced membrane-bounded vesicles of the columnar cells of the rat intestinal epithelium during fat absorption. J Ultrastruct Res. 1967 Sep;20(1):140–160. doi: 10.1016/s0022-5320(67)80042-x. [DOI] [PubMed] [Google Scholar]
  41. Strauss E. W. Electron microscopic study of intestinal fat absorption in vitro from mixed micelles containing linolenic acid, monoolein, and bile salt. J Lipid Res. 1966 Mar;7(2):307–323. [PubMed] [Google Scholar]
  42. WATSON M. L. Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol. 1958 Jul 25;4(4):475–478. doi: 10.1083/jcb.4.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. WEISS J. M. The role of the Golgi complex in fat absorption as studied with the electron microscope with observations on the cytology of duodenal absorptive cells. J Exp Med. 1955 Dec 1;102(6):775–782. doi: 10.1084/jem.102.6.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weinstock A. Cytotoxic effects of puromycin on the Golgi apparatus of pancreatic acinar cells, hepatocytes and ameloblasts. J Histochem Cytochem. 1970 Dec;18(12):875–886. doi: 10.1177/18.12.875. [DOI] [PubMed] [Google Scholar]
  45. Windmueller H. G., Levy R. I. Production of beta-lipoprotein by intestine in the rat. J Biol Chem. 1968 Sep 25;243(18):4878–4884. [PubMed] [Google Scholar]
  46. Wise G. E., Flickinger C. J. Cytochemical staining of the Golgi apparatus in amoeba proteus. J Cell Biol. 1970 Sep;46(3):620–626. doi: 10.1083/jcb.46.3.620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. ZEIGEL R. F., DALTON A. J. Speculations based on the morphology of the Golgi systems in several types of proteinsecreting cells. J Cell Biol. 1962 Oct;15:45–54. doi: 10.1083/jcb.15.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. de Groot N., Panet A., Lapidot Y. Reaction of puromycin with chemically prepared peptidyl transfer RNA. Eur J Biochem. 1970 Aug;15(2):215–221. doi: 10.1111/j.1432-1033.1970.tb00997.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES