Skip to main content
The Journal of Physiology logoLink to The Journal of Physiology
. 1976 Jun;257(3):561–579. doi: 10.1113/jphysiol.1976.sp011385

Mode of stimulation by adenosine 3':5'-cyclic monophosphate of the sodium efflux in barnacle muscle fibres.

E E Bittar, G Chambers, R Schultz
PMCID: PMC1309379  PMID: 181561

Abstract

1. Giant fibres of the barnacle Balanus nubilus have been used as a preparation for studying the mode of action of cAMP on sodium transport. 2. It is shown that a concentration of cAMP as low as 10(-6)M, when micro-injected, causes a sharp rise in the radio-Na efflux. Ouabain fails to reverse the cAMP effect. 3. The magnitude of the response of the Na efflux to cAMP is markedly reduced by pre-injecting 100 or 500 mM-EGTA solutions or by omitting Ca2+ from the bathing medium. Both together fail to bring about a greater reduction in the response. 4. The response to cAMP is greatly reduced by pre-injecting the protein inhibitor of Walsh and practically abolished by pre-injecting 500 mM-EGTA and soaking in Ca-free artificial sea water, ASW. 5. The Ca2+-independent component of the Na efflux which is also stimulated by cAMP is shown to involve Na for H exchange. The magnitude of this exchange is governed by external pH. 6. The Na efflux into Ca2+-free, Li+-ASW is shown to be markedly stimulated by injecting cAMP, an effect which is enhanced by reducing external pH. 7. The Na efflux at 0 degrees C is stimulated by injecting cAMP. This is shown to be related to activation of the protein kinase by cAMP and to depend on the presence of external Ca2+. 8 (i) Ethacrynic acid when injected reduces the ouabain-insensitive Na efflux into HEPES-Ca2+-free ASW at pH 6-3. These same fibres show a marked response to cAMP. (II) The ouabain-insensitive Na efflux into HCO3-, Ca2+-free ASW from fibres pre-treated with ethacrynic acid fails to respond to external acidification. This is interpreted as indicating that ethacrynic acid inactivates the CO2-sensitive adenyl cyclase system. These same fibres when injected with cAMP show a marked response. (iii) Stimulation of the ouabain-insensitive Na efflux into HCO-3, Ca2+-free ASW by external acidification is reversed by injecting ethacrynic acid. These fibres when injected with cAMP show a reduced response. 9. It is concluded that: (i) stimulation of the Na efflux by injected cAMP is mainly due to activation of cAMP-dependent protein kinase; (ii) the underlying exchange mechanism consists of Na:Ca and Na:H exchange. Interaction of Ca2+ with a phosphorylated membrane, thereby modifying permeability remains as a real possibility; (iii) the site of action of CO2 and ethacrynic acid is the adenyl cyclase system. 10. The implications of activation of the adenyl cyclase system by CO2 and Na:H exchange are briefly touched upon.

Full text

PDF
561

Selected References

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

  1. Ashby C. D., Walsh D. A. Characterization of the interaction of a protein inhibitor with adenosine 3',5'-monophosphate-dependent protein kinases. I. Interaction with the catalytic subunit of the protein kinase. J Biol Chem. 1972 Oct 25;247(20):6637–6642. [PubMed] [Google Scholar]
  2. Ashby C. D., Walsh D. A. Characterization of the interaction of a protein inhibitor with adenosine 3',5'-monophosphate-dependent protein kinases. II. Mechanism of action with the holoenzyme. J Biol Chem. 1973 Feb 25;248(4):1255–1261. [PubMed] [Google Scholar]
  3. Beavo J. A., Bechtel P. J., Krebs E. G. Activation of protein kinase by physiological concentrations of cyclic AMP. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3580–3583. doi: 10.1073/pnas.71.9.3580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beavo J. A., Bechtel P. J., Krebs E. G. Mechanisms of control for cAMP-dependent protein kinase from skeletal muscle. Adv Cyclic Nucleotide Res. 1975;5:241–251. [PubMed] [Google Scholar]
  5. Bittar E. E., Chen S., Danielson B. G., Hartmann H. A., Tong E. Y. An investigation of sodium transport in barnacle muscle fibres by means of the microsyringe technique. J Physiol. 1972 Mar;221(2):389–414. doi: 10.1113/jphysiol.1972.sp009757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bittar E. E. Effect of inhibitors and uncouplers on the Na pump of the Maia muscle fibre. J Physiol. 1966 Nov;187(1):81–103. doi: 10.1113/jphysiol.1966.sp008077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bittar E. E., Hift H., Huddart H., Tong E. The effects of caffeine on sodium transport, membrane potential, mechanical tension and ultrastructure in barnacle muscle fibres. J Physiol. 1974 Oct;242(1):1–34. doi: 10.1113/jphysiol.1974.sp010691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bittar E. E., Tallitsch R. B. Mode of stimulation by aldosterone of the sodium efflux in barnacle muscle fibres: effects of ouabain, ethacrynic acid, diphenylhydantoin, (ATPMg)(2-), adenine translocase inhibitors, pyruvate and oxythiamine. J Physiol. 1976 Feb;255(1):29–56. doi: 10.1113/jphysiol.1976.sp011268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bittar E. E., Tong E. Sensitivity of the sodium efflux in barnacle muscle fibers to the microinjection of ATP. Life Sci. 1975 Jan 15;16(2):289–296. doi: 10.1016/0024-3205(75)90027-2. [DOI] [PubMed] [Google Scholar]
  10. Borle A. B. Cyclic AMP stimulation of calcium efflux from kidney, liver and heart mitochondria. J Membr Biol. 1974;16(3):221–236. doi: 10.1007/BF01872416. [DOI] [PubMed] [Google Scholar]
  11. CALDWELL P. C., WALSTER G. STUDIES ON THE MICRO-INJECTION OF VARIOUS SUBSTANCES INTO CRAB MUSCLE FIBRES. J Physiol. 1963 Nov;169:353–372. doi: 10.1113/jphysiol.1963.sp007261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Danielson B. G., Bittar E. E., Chen S. S., Tong E. Y. Inhibition of the CO 2 -sensitive Na efflux in barnacle muscle fibers by micro-injection of ethacrynic acid. Experientia. 1972 Nov 15;28(11):1304–1304. doi: 10.1007/BF01965309. [DOI] [PubMed] [Google Scholar]
  13. Danielson B. G., Bittar E. E., Chen S. S., Tong E. Y. On the mode of action of ethacrynic acid, using the barnacle muscle fiber as a model. Life Sci I. 1972 Jan 1;11(1):13–21. doi: 10.1016/0024-3205(72)90237-8. [DOI] [PubMed] [Google Scholar]
  14. Danielson B. G., Bittar E. E., Chen S., Tong E. The influence of low pH, high K and microinjected CaCl 2 on the ouabain-insensitive component of sodium efflux in barnacle muscle fibers. Life Sci I. 1971 Jul 15;10(14):833–839. doi: 10.1016/0024-3205(71)90038-5. [DOI] [PubMed] [Google Scholar]
  15. Ebel H. Effect of diuretics on renal NaK-ATPase and adenyl cyclase. Naunyn Schmiedebergs Arch Pharmacol. 1974;281(3):301–314. doi: 10.1007/BF00500599. [DOI] [PubMed] [Google Scholar]
  16. Entman M. L. The role of cyclic AMP in the modulation of cardiac contractility. Adv Cyclic Nucleotide Res. 1974;4(0):163–193. [PubMed] [Google Scholar]
  17. Gordon E. E., de Hartog M. The relationship between cell membrane potassium ion transport and glycolysis. The effect of ethacrynic acid. J Gen Physiol. 1969 Nov;54(5):650–663. doi: 10.1085/jgp.54.5.650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. HODGKIN A. L., KEYNES R. D. Experiments on the injection of substances into squid giant axons by means of a microsyringe. J Physiol. 1956 Mar 28;131(3):592–616. doi: 10.1113/jphysiol.1956.sp005485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Haddox M. K., Newton N. E., Hartle D. K., Goldberg N. D. ATP(Mg 2+ ) induced inhibition of cyclic AMP reactivity with a skeletal muscle protein kinase. Biochem Biophys Res Commun. 1972 May 26;47(4):653–661. doi: 10.1016/0006-291x(72)90542-6. [DOI] [PubMed] [Google Scholar]
  20. Heilmeyer L. M., Jr, Meyer F., Haschke R. H., Fischer E. H. Control of phosphorylase activity in a muscle glycogen particle. II. Activation by calcium. J Biol Chem. 1970 Dec 25;245(24):6649–6656. [PubMed] [Google Scholar]
  21. Klahr S., Yates J., Bourgoignie J. Inhibition of glycolysis by ethacrynic acid and furosemide. Am J Physiol. 1971 Oct;221(4):1038–1043. doi: 10.1152/ajplegacy.1971.221.4.1038. [DOI] [PubMed] [Google Scholar]

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

RESOURCES