Skip to main content
NCBI home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1997 Nov 1;504(Pt 3):695–704. doi: 10.1111/j.1469-7793.1997.695bd.x

Adenosine formation in contracting primary rat skeletal muscle cells and endothelial cells in culture.

Y Hellsten 1, U Frandsen 1
PMCID: PMC1159971  PMID: 9401975

Abstract

1. The present study examined the capacity for adenosine formation, uptake and metabolism in contracting primary rat muscle cells and in microvascular endothelial cells in culture. 2. Strong and moderate electrical simulation of skeletal muscle cells led to a significantly greater increase in the extracellular adenosine concentration (421 +/- 91 and 235 +/- 30 nmol (g protein)-1, respectively; P < 0.05) compared with non-stimulated muscle cells (161 +/- 20 nmol (g protein)-1). The ATP concentration was lower (18%; P < 0.05) in the intensely contracted, but not in the moderately contracted muscle cells. 3. Addition of microvascular endothelial cells to the cultured skeletal muscle cells enhanced the contraction-induced accumulation of extracellular adenosine (P < 0.05), whereas endothelial cells in culture alone did not cause extracellular accumulation of adenosine. 4. Skeletal muscle cells were found to have ecto-forms of several enzymes involved in nucleotide metabolism, including ATPases capable of converting extracellular ATP to ADP and AMP. 5. Adenosine added to the cell medium was taken up by muscle cells and incorporated into the adenine nucleotide pool so that after 30 min of incubation, over 95% of the adenosine label was present in ATP, ADP and AMP. A similar extent of incorporation of adenosine into the nucleotide pool was evident in the endothelial cells. 6. The present data suggest that contracting muscle cells induce an elevation in the extracellular adenosine concentration. Addition of endothelial cells to muscle cells enhances the contraction-induced formation of adenosine. Adenosine taken up by muscle and endothelial cells from the extracellular space is not likely to be used for storage in intracellular pools, but may serve to regulate muscle extracellular adenosine levels.

Full text

PDF
695

Selected References

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

  1. Andersen P., Saltin B. Maximal perfusion of skeletal muscle in man. J Physiol. 1985 Sep;366:233–249. doi: 10.1113/jphysiol.1985.sp015794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arabadjis P. G., Tullson P. C., Terjung R. L. Purine nucleoside formation in rat skeletal muscle fiber types. Am J Physiol. 1993 May;264(5 Pt 1):C1246–C1251. doi: 10.1152/ajpcell.1993.264.5.C1246. [DOI] [PubMed] [Google Scholar]
  3. Bangsbo J., Gollnick P. D., Graham T. E., Juel C., Kiens B., Mizuno M., Saltin B. Anaerobic energy production and O2 deficit-debt relationship during exhaustive exercise in humans. J Physiol. 1990 Mar;422:539–559. doi: 10.1113/jphysiol.1990.sp018000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bockman E. L., McKenzie J. E. Tissue adenosine content in active soleus and gracilis muscles of cats. Am J Physiol. 1983 Apr;244(4):H552–H559. doi: 10.1152/ajpheart.1983.244.4.H552. [DOI] [PubMed] [Google Scholar]
  5. Burger R. M., Lowenstein J. M. Preparation and properties of 5'-nucleotidase from smooth muscle of small intestine. J Biol Chem. 1970 Dec 10;245(23):6274–6280. [PubMed] [Google Scholar]
  6. Cunha R. A., Sebastião A. M. Adenosine and adenine nucleotides are independently released from both the nerve terminals and the muscle fibres upon electrical stimulation of the innervated skeletal muscle of the frog. Pflugers Arch. 1993 Sep;424(5-6):503–510. doi: 10.1007/BF00374914. [DOI] [PubMed] [Google Scholar]
  7. Daniels M. P. Localization of actin, beta-spectrin, 43 x 10(3) Mr and 58 x 10(3) Mr proteins to receptor-enriched domains of newly formed acetylcholine receptor aggregates in isolated myotube membranes. J Cell Sci. 1990 Dec;97(Pt 4):615–626. doi: 10.1242/jcs.97.4.615. [DOI] [PubMed] [Google Scholar]
  8. Deussen A., Möser G., Schrader J. Contribution of coronary endothelial cells to cardiac adenosine production. Pflugers Arch. 1986 Jun;406(6):608–614. doi: 10.1007/BF00584028. [DOI] [PubMed] [Google Scholar]
  9. Dobson J. G., Jr, Rubio R., Berne R. M. Role of adenine nucleotides, adenosine, and inorganic phosphate in the regulation of skeletal muscle blood flow. Circ Res. 1971 Oct;29(4):375–384. doi: 10.1161/01.res.29.4.375. [DOI] [PubMed] [Google Scholar]
  10. Drury A. N., Szent-Györgyi A. The physiological activity of adenine compounds with especial reference to their action upon the mammalian heart. J Physiol. 1929 Nov 25;68(3):213–237. doi: 10.1113/jphysiol.1929.sp002608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Edlund A., Sollevi A., Linde B. Haemodynamic and metabolic effects of infused adenosine in man. Clin Sci (Lond) 1990 Aug;79(2):131–138. doi: 10.1042/cs0790131. [DOI] [PubMed] [Google Scholar]
  12. Gordon E. L., Pearson J. D., Slakey L. L. The hydrolysis of extracellular adenine nucleotides by cultured endothelial cells from pig aorta. Feed-forward inhibition of adenosine production at the cell surface. J Biol Chem. 1986 Nov 25;261(33):15496–15507. [PubMed] [Google Scholar]
  13. Jacobs A. E., Oosterhof A., Veerkamp J. H. Purine and pyrimidine metabolism in human muscle and cultured muscle cells. Biochim Biophys Acta. 1988 Jun 30;970(2):130–136. doi: 10.1016/0167-4889(88)90171-1. [DOI] [PubMed] [Google Scholar]
  14. Nees S., Herzog V., Becker B. F., Böck M., Des Rosiers Ch, Gerlach E. The coronary endothelium: a highly active metabolic barrier for adenosine. Basic Res Cardiol. 1985 Sep-Oct;80(5):515–529. doi: 10.1007/BF01907915. [DOI] [PubMed] [Google Scholar]
  15. Pearson J. D., Carleton J. S., Hutchings A., Gordon J. L. Uptake and metabolism of adenosine by pig aortic endothelial and smooth-muscle cells in culture. Biochem J. 1978 Feb 15;170(2):265–271. doi: 10.1042/bj1700265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rubio R., Berne R. M., Dobson J. G., Jr Sites of adenosine production in cardiac and skeletal muscle. Am J Physiol. 1973 Oct;225(4):938–953. doi: 10.1152/ajplegacy.1973.225.4.938. [DOI] [PubMed] [Google Scholar]
  17. Ryan J. W., Smith U. Metabolism of adenosine 5'-monophosphate during circulation through the lungs. Trans Assoc Am Physicians. 1971;84:297–306. [PubMed] [Google Scholar]
  18. Schrader J., Berne R. M., Rubio R. Uptake and metabolism of adenosine by human erythrocyte ghosts. Am J Physiol. 1972 Jul;223(1):159–166. doi: 10.1152/ajplegacy.1972.223.1.159. [DOI] [PubMed] [Google Scholar]
  19. Tullson P. C., Bangsbo J., Hellsten Y., Richter E. A. IMP metabolism in human skeletal muscle after exhaustive exercise. J Appl Physiol (1985) 1995 Jan;78(1):146–152. doi: 10.1152/jappl.1995.78.1.146. [DOI] [PubMed] [Google Scholar]
  20. Tullson P. C., Whitlock D. M., Terjung R. L. Adenine nucleotide degradation in slow-twitch red muscle. Am J Physiol. 1990 Feb;258(2 Pt 1):C258–C265. doi: 10.1152/ajpcell.1990.258.2.C258. [DOI] [PubMed] [Google Scholar]
  21. Voyta J. C., Via D. P., Butterfield C. E., Zetter B. R. Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J Cell Biol. 1984 Dec;99(6):2034–2040. doi: 10.1083/jcb.99.6.2034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wagner R. C., Matthews M. A. The isolation and culture of capillary endothelium from epididymal fat. Microvasc Res. 1975 Nov;10(3):286–297. doi: 10.1016/0026-2862(75)90033-3. [DOI] [PubMed] [Google Scholar]
  23. Zoref-Shani E., Shainberg A., Sperling O. Pathways of adenine nucleotide catabolism in primary rat muscle cultures. Biochim Biophys Acta. 1987 Dec 7;926(3):287–295. doi: 10.1016/0304-4165(87)90215-7. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES