Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1985 Nov 1;231(3):713–719. doi: 10.1042/bj2310713

Fuel utilization in colonocytes of the rat.

M S Ardawi, E A Newsholme
PMCID: PMC1152807  PMID: 4074334

Abstract

In incubated colonocytes isolated from rat colons, the rates of utilization O2, glucose or glutamine were linear with respect to time for over 30 min, and the concentrations of adenine nucleotides plus the ATP/ADP or ATP/AMP concentration ratios remained approximately constant for 30 min. Glutamine, n-butyrate or ketone bodies were the only substrates that caused increases in O2 consumption by isolated incubated colonocytes. The maximum activity of hexokinase in colonic mucosa is similar to that of 6-phosphofructokinase. Starvation of the donor animal decreased the activities of hexokinase and 6-phosphofructokinase, whereas it increased those of glucose-6-phosphatase and fructose-bisphosphatase. Isolated incubated colonocytes utilized glucose at about 6.8 mumol/min per g dry wt., with lactate accounting for 83% of glucose removed. These rates were not affected by the addition of glutamine, acetoacetate or n-butyrate, and starvation of the donor animal. Isolated incubated colonocytes utilized glutamine at about 5.5 mumol/min per g dry wt., which is about 21% of the maximum activity of glutaminase. The major end-products of glutamine metabolism were glutamate, aspartate, alanine and ammonia. Starvation of the donor animal decreased the rate of glutamine utilization by colonocytes, which is accompanied by a decrease in glutamate formation and in the maximum activity of glutaminase. Isolated incubated colonocytes utilized acetoacetate at about 3.5 mumol/min per g dry wt. This rate was not markedly affected by addition of glucose or by starvation of the donor animal. When colonocytes were incubated with n-butyrate, both acetoacetate and 3-hydroxybutyrate were formed, with the latter accounting for only about 19% of total ketones produced.

Full text

PDF
713

Selected References

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

  1. Alp P. R., Newsholme E. A., Zammit V. A. Activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenase in muscle from vertebrates and invertebrates. Biochem J. 1976 Mar 15;154(3):689–700. doi: 10.1042/bj1540689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson J. W. Glucose metabolism in jejunal mucosa of fed, fasted, and streptozotocin-diabetic rats. Am J Physiol. 1974 Jan;226(1):226–229. doi: 10.1152/ajplegacy.1974.226.1.226. [DOI] [PubMed] [Google Scholar]
  3. Ardawi M. S., Newsholme E. A. Glutamine metabolism in lymphocytes of the rat. Biochem J. 1983 Jun 15;212(3):835–842. doi: 10.1042/bj2120835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ardawi M. S., Newsholme E. A. Maximum activities of some enzymes of glycolysis, the tricarboxylic acid cycle and ketone-body and glutamine utilization pathways in lymphocytes of the rat. Biochem J. 1982 Dec 15;208(3):743–748. doi: 10.1042/bj2080743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ardawi M. S., Newsholme E. A. Metabolism of ketone bodies, oleate and glucose in lymphocytes of the rat. Biochem J. 1984 Jul 1;221(1):255–260. doi: 10.1042/bj2210255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brand K., Williams J. F., Weidemann M. J. Glucose and glutamine metabolism in rat thymocytes. Biochem J. 1984 Jul 15;221(2):471–475. doi: 10.1042/bj2210471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Budohoski L., Challis R. A., Newsholme E. A. Effects of starvation on the maximal activities of some glycolytic and citric acid-cycle enzymes and glutaminase in mucosa of the small intestine of the rat. Biochem J. 1982 Jul 15;206(1):169–172. doi: 10.1042/bj2060169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cooney G. J., Taegtmeyer H., Newsholme E. A. Tricarboxylic acid cycle flux and enzyme activities in the isolated working rat heart. Biochem J. 1981 Dec 15;200(3):701–703. doi: 10.1042/bj2000701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crabtree B., Higgins S. J., Newsholme E. A. The activities of pyruvate carboxylase, phosphoenolpyruvate carboxylase and fructose diphosphatase in muscles from vertebrates and invertebrates. Biochem J. 1972 Nov;130(2):391–396. doi: 10.1042/bj1300391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Crabtree B., Newsholme E. A. The activities of phosphorylase, hexokinase, phosphofructokinase, lactate dehydrogenase and the glycerol 3-phosphate dehydrogenases in muscles from vertebrates and invertebrates. Biochem J. 1972 Jan;126(1):49–58. doi: 10.1042/bj1260049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Culvenor J. G., Weidemann M. J. Phytohaemagglutinin stimulation of rat thymus lymphocytes glycolysis. Biochim Biophys Acta. 1976 Jul 21;437(2):354–363. doi: 10.1016/0304-4165(76)90005-2. [DOI] [PubMed] [Google Scholar]
  12. Curthoys N. P., Lowry O. H. The distribution of glutaminase isoenzymes in the various structures of the nephron in normal, acidotic, and alkalotic rat kidney. J Biol Chem. 1973 Jan 10;248(1):162–168. [PubMed] [Google Scholar]
  13. Denton R. M., Halestrap A. P. Regulation of pyruvate metabolism in mammalian tissues. Essays Biochem. 1979;15:37–77. [PubMed] [Google Scholar]
  14. Hanson P. J., Carrington J. M. Activity of 3-oxo acid CoA-transferase, D-3-hydroxybutyrate dehydrogenase, hexokinase and carnitine palmitoyltransferase in the stomach and small and large intestine of the rat. Biochem J. 1981 Nov 15;200(2):349–355. doi: 10.1042/bj2000349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hanson P. J., Parsons D. S. Factors affecting the utilization of ketone bodies and other substrates by rat jejunum: effects of fasting and of diabetes. J Physiol. 1978 May;278:55–67. doi: 10.1113/jphysiol.1978.sp012292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hanson P. J., Parsons D. S. The interrelationship between glutamine and alanine in the intestine. Biochem Soc Trans. 1980 Oct;8(5):506–509. doi: 10.1042/bst0080506. [DOI] [PubMed] [Google Scholar]
  17. Hanson P. J., Parsons S. Metabolism and transport of glutamine and glucose in vascularly perfused small intestine rat. Biochem J. 1977 Sep 15;166(3):509–519. doi: 10.1042/bj1660509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Henning S. J., Hird F. J. Ketogenesis from butyrate and acetate by the caecum and the colon of rabbits. Biochem J. 1972 Dec;130(3):785–790. doi: 10.1042/bj1300785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hume D. A., Radik J. L., Ferber E., Weidemann M. J. Aerobic glycolysis and lymphocyte transformation. Biochem J. 1978 Sep 15;174(3):703–709. doi: 10.1042/bj1740703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Lackner R., Challiss R. A., West D., Newsholme E. A. A problem in the radiochemical assay of glucose-6-phosphatase in muscle. Biochem J. 1984 Mar 1;218(2):649–651. doi: 10.1042/bj2180649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Opie L. H., Newsholme E. A. The inhibition of skeletal-muscle fructose 1,6-diphosphatase by adenosine monophosphate. Biochem J. 1967 Aug;104(2):353–360. doi: 10.1042/bj1040353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Porteous J. W. Glutamate, glutamine, aspartate, asparagine, glucose and ketone-body metabolism in chick intestinal brush-border cells. Biochem J. 1980 Jun 15;188(3):619–632. doi: 10.1042/bj1880619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Read G., Crabtree B., Smith G. H. The activities of 2-oxoglutarate dehydrogenase and pyruvate dehydrogenase in hearts and mammary glands from ruminants and non-ruminants. Biochem J. 1977 May 15;164(2):349–355. doi: 10.1042/bj1640349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Roediger W. E., Truelove S. C. Method of preparing isolated colonic epithelial cells (colonocytes) for metabolic studies. Gut. 1979 Jun;20(6):484–488. doi: 10.1136/gut.20.6.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Roediger W. E. Utilization of nutrients by isolated epithelial cells of the rat colon. Gastroenterology. 1982 Aug;83(2):424–429. [PubMed] [Google Scholar]
  27. Sugden P. H., Newsholme E. A. Activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenases, glutamate dehydrogenase, aspartate aminotransferase and alanine aminotransferase in nervous tissues from vertebrates and invertebrates. Biochem J. 1975 Jul;150(1):105–111. doi: 10.1042/bj1500105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vinay P., Lemieux G., Gougoux A., Lemieux C. Response of the rat and dog kidney to H+ concentration in vitro--a comparative study with slices and tubules. Int J Biochem. 1980;12(1-2):89–98. doi: 10.1016/0020-711x(80)90048-8. [DOI] [PubMed] [Google Scholar]
  29. WOLLENBERGER A., RISTAU O., SCHOFFA G. [A simple technic for extremely rapid freezing of large pieces of tissue]. Pflugers Arch Gesamte Physiol Menschen Tiere. 1960;270:399–412. [PubMed] [Google Scholar]
  30. Watford M., Lund P., Krebs H. A. Isolation and metabolic characteristics of rat and chicken enterocytes. Biochem J. 1979 Mar 15;178(3):589–596. doi: 10.1042/bj1780589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Watford M., Smith E. M., Erbelding E. J. The regulation of phosphate-activated glutaminase activity and glutamine metabolism in the streptozotocin-diabetic rat. Biochem J. 1984 Nov 15;224(1):207–214. doi: 10.1042/bj2240207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Williamson D. H., Bates M. W., Page M. A., Krebs H. A. Activities of enzymes involved in acetoacetate utilization in adult mammalian tissues. Biochem J. 1971 Jan;121(1):41–47. doi: 10.1042/bj1210041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Williamson D. H., Lund P., Krebs H. A. The redox state of free nicotinamide-adenine dinucleotide in the cytoplasm and mitochondria of rat liver. Biochem J. 1967 May;103(2):514–527. doi: 10.1042/bj1030514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Windmueller H. G., Spaeth A. E. Identification of ketone bodies and glutamine as the major respiratory fuels in vivo for postabsorptive rat small intestine. J Biol Chem. 1978 Jan 10;253(1):69–76. [PubMed] [Google Scholar]
  35. Windmueller H. G., Spaeth A. E. Respiratory fuels and nitrogen metabolism in vivo in small intestine of fed rats. Quantitative importance of glutamine, glutamate, and aspartate. J Biol Chem. 1980 Jan 10;255(1):107–112. [PubMed] [Google Scholar]
  36. Windmueller H. G., Spaeth A. E. Uptake and metabolism of plasma glutamine by the small intestine. J Biol Chem. 1974 Aug 25;249(16):5070–5079. [PubMed] [Google Scholar]
  37. Yasmeen D., Laird A. J., Hume D. A., Weidemann M. J. Activation of 3-O-methyl-glucose transport in rat thymus lymphocytes by concanavalin A. Temperature and calcium ion dependence and sensitivity to puromycin but to cycloheximide. Biochim Biophys Acta. 1977 Nov 7;500(1):89–102. doi: 10.1016/0304-4165(77)90049-6. [DOI] [PubMed] [Google Scholar]
  38. Zammit V. A., Beis I., Newsholme E. A. Maximum activities and effects of fructose bisphosphate on pyruvate kinase from muscles of vertebrates and invertebrates in relation to the control of glycolysis. Biochem J. 1978 Sep 15;174(3):989–998. doi: 10.1042/bj1740989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES