Skip to main content
PMC home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1996 Oct 1;496(Pt 1):265–273. doi: 10.1113/jphysiol.1996.sp021683

Interactions between secretin and acetylcholine in the regulation of fluid secretion by isolated rat pancreatic ducts.

R L Evans 1, N Ashton 1, A C Elliott 1, R Green 1, B E Argent 1
PMCID: PMC1160842  PMID: 8910214

Abstract

1. Interlobular ducts were isolated from the rat pancreas and maintained in short-term tissue culture. Fluid secretion from these isolated ducts was measured using micropuncture techniques, intracellular calcium concentration ([Ca2+]i) by fura-2 microspectrofluorimetry, and cyclic AMP by radioimmunoassay. 2. Applying secretin and ACh simultaneously to ducts caused either a stimulation or an inhibition of fluid secretion depending on the doses employed. 3. The inhibitory effect of secretin and ACh could be relieved by atropine, and by the protein kinase C (PKC) inhibitors staurosporine and 1-(5-isoquinolinylsulphonyl)-2-methyl-piperazine (H-7). 4. Activation of PKC by 12-O-tetradecanoylphorbol-13-acetate (TPA) and phorbol 12, 13-dibutyrate (PDBu) inhibited secretin-evoked fluid secretion. 5. ACh and TPA also inhibited fluid secretion stimulated by the adenylate cyclase activator, forskolin. 6. Neither secretin nor the PKC activators and inhibitors had any effect on either the increase in [Ca2+]i evoked by ACh or the increase in intracellular cyclic AMP evoked by secretin and forskolin. 7. We conclude that the inhibitory effect of combined doses of secretin and ACh on ductal fluid secretion is probably mediated by PKC at a point in the secretory mechanism distal to the generation of intracellular messengers.

Full text

PDF
265

Selected References

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

  1. Argent B. E., Arkle S., Cullen M. J., Green R. Morphological, biochemical and secretory studies on rat pancreatic ducts maintained in tissue culture. Q J Exp Physiol. 1986 Oct;71(4):633–648. doi: 10.1113/expphysiol.1986.sp003023. [DOI] [PubMed] [Google Scholar]
  2. Arkle S., Lee C. M., Cullen M. J., Argent B. E. Isolation of ducts from the pancreas of copper-deficient rats. Q J Exp Physiol. 1986 Apr;71(2):249–265. doi: 10.1113/expphysiol.1986.sp002982. [DOI] [PubMed] [Google Scholar]
  3. Ashton N., Argent B. E., Green R. Characteristics of fluid secretion from isolated rat pancreatic ducts stimulated with secretin and bombesin. J Physiol. 1991 Apr;435:533–546. doi: 10.1113/jphysiol.1991.sp018523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ashton N., Argent B. E., Green R. Effect of vasoactive intestinal peptide, bombesin and substance P on fluid secretion by isolated rat pancreatic ducts. J Physiol. 1990 Aug;427:471–482. doi: 10.1113/jphysiol.1990.sp018182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ashton N., Evans R. L., Elliott A. C., Green R., Argent B. E. Regulation of fluid secretion and intracellular messengers in isolated rat pancreatic ducts by acetylcholine. J Physiol. 1993 Nov;471:549–562. doi: 10.1113/jphysiol.1993.sp019915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chabardès D., Montégut M., Imbert-Teboul M., Morel F. Inhibition of alpha 2-adrenergic agonists on AVP-induced cAMP accumulation in isolated collecting tubule of the rat kidney. Mol Cell Endocrinol. 1984 Oct;37(3):263–275. doi: 10.1016/0303-7207(84)90096-0. [DOI] [PubMed] [Google Scholar]
  7. Fölsch U. R., Fischer H., Söling H. D., Creutzfeldt W. Effects of gastrointestinal hormones and carbamylcholine on cAMP accumulation in isolated pancreatic duct fragments from the rat. Digestion. 1980;20(4):277–292. doi: 10.1159/000198449. [DOI] [PubMed] [Google Scholar]
  8. Gray M. A., Greenwell J. R., Argent B. E. Secretin-regulated chloride channel on the apical plasma membrane of pancreatic duct cells. J Membr Biol. 1988 Oct;105(2):131–142. doi: 10.1007/BF02009166. [DOI] [PubMed] [Google Scholar]
  9. Gray M. A., Greenwell J. R., Garton A. J., Argent B. E. Regulation of maxi-K+ channels on pancreatic duct cells by cyclic AMP-dependent phosphorylation. J Membr Biol. 1990 May;115(3):203–215. doi: 10.1007/BF01868636. [DOI] [PubMed] [Google Scholar]
  10. Gray M. A., Harris A., Coleman L., Greenwell J. R., Argent B. E. Two types of chloride channel on duct cells cultured from human fetal pancreas. Am J Physiol. 1989 Aug;257(2 Pt 1):C240–C251. doi: 10.1152/ajpcell.1989.257.2.C240. [DOI] [PubMed] [Google Scholar]
  11. Gray M. A., Plant S., Argent B. E. cAMP-regulated whole cell chloride currents in pancreatic duct cells. Am J Physiol. 1993 Mar;264(3 Pt 1):C591–C602. doi: 10.1152/ajpcell.1993.264.3.C591. [DOI] [PubMed] [Google Scholar]
  12. Gray M. A., Winpenny J. P., Porteous D. J., Dorin J. R., Argent B. E. CFTR and calcium-activated chloride currents in pancreatic duct cells of a transgenic CF mouse. Am J Physiol. 1994 Jan;266(1 Pt 1):C213–C221. doi: 10.1152/ajpcell.1994.266.1.C213. [DOI] [PubMed] [Google Scholar]
  13. Habara Y. Influences of ionic environments on ACh-induced secretory responses in isolated perfused pancreas of rats. Jpn J Physiol. 1980;30(4):561–574. doi: 10.2170/jjphysiol.30.561. [DOI] [PubMed] [Google Scholar]
  14. Helmle-Kolb C., Montrose M. H., Stange G., Murer H. Regulation of Na+/H+ exchange in opossum kidney cells by parathyroid hormone, cyclic AMP and phorbol esters. Pflugers Arch. 1990 Jan;415(4):461–470. doi: 10.1007/BF00373624. [DOI] [PubMed] [Google Scholar]
  15. Hidaka H., Inagaki M., Kawamoto S., Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry. 1984 Oct 9;23(21):5036–5041. doi: 10.1021/bi00316a032. [DOI] [PubMed] [Google Scholar]
  16. Hootman S. R., Zukerman J., Kovalcik S. A. Muscarinic receptors in isolated guinea pig pancreatic ducts. Biochem Pharmacol. 1993 Jul 20;46(2):291–296. doi: 10.1016/0006-2952(93)90417-u. [DOI] [PubMed] [Google Scholar]
  17. Hug M., Pahl C., Novak I. Effect of ATP, carbachol and other agonists on intracellular calcium activity and membrane voltage of pancreatic ducts. Pflugers Arch. 1994 Mar;426(5):412–418. doi: 10.1007/BF00388304. [DOI] [PubMed] [Google Scholar]
  18. Kase H., Wakui M., Petersen O. H. Stimulatory and inhibitory actions of VIP and cyclic AMP on cytoplasmic Ca2+ signal generation in pancreatic acinar cells. Pflugers Arch. 1991 Dec;419(6):668–670. doi: 10.1007/BF00370314. [DOI] [PubMed] [Google Scholar]
  19. Madden M. E., Sarras M. P., Jr The pancreatic ductal system of the rat: cell diversity, ultrastructure, and innervation. Pancreas. 1989;4(4):472–485. doi: 10.1097/00006676-198908000-00013. [DOI] [PubMed] [Google Scholar]
  20. Maruyama Y. Activation and desensitization mechanisms of muscarinic current response in single pancreatic acinar cells of rats. J Physiol. 1989 Oct;417:343–359. doi: 10.1113/jphysiol.1989.sp017805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matthews J. B., Awtrey C. S., Hecht G., Tally K. J., Thompson R. S., Madara J. L. Phorbol ester sequentially downregulates cAMP-regulated basolateral and apical Cl- transport pathways in T84 cells. Am J Physiol. 1993 Oct;265(4 Pt 1):C1109–C1117. doi: 10.1152/ajpcell.1993.265.4.C1109. [DOI] [PubMed] [Google Scholar]
  22. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  23. Novak I., Greger R. Electrophysiological study of transport systems in isolated perfused pancreatic ducts: properties of the basolateral membrane. Pflugers Arch. 1988 Jan;411(1):58–68. doi: 10.1007/BF00581647. [DOI] [PubMed] [Google Scholar]
  24. Novak I., Greger R. Properties of the luminal membrane of isolated perfused rat pancreatic ducts. Effect of cyclic AMP and blockers of chloride transport. Pflugers Arch. 1988 May;411(5):546–553. doi: 10.1007/BF00582376. [DOI] [PubMed] [Google Scholar]
  25. Pahl C., Novak I. Effect of vasoactive intestinal peptide, carbachol and other agonists on the membrane voltage of pancreatic duct cells. Pflugers Arch. 1993 Aug;424(3-4):315–320. doi: 10.1007/BF00384358. [DOI] [PubMed] [Google Scholar]
  26. Petersen O. H., Ueda N. Secretion of fluid and amylase in the perfused rat pancreas. J Physiol. 1977 Jan;264(3):819–835. doi: 10.1113/jphysiol.1977.sp011696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Salido G. M., Francis L. P., Camello P. J., Singh J., Madrid J. A., Pariente J. A. Effects of phorbol esters and secretin on pancreatic juice secretion in the anaesthetized rat. Gen Pharmacol. 1990;21(4):465–469. doi: 10.1016/0306-3623(90)90699-m. [DOI] [PubMed] [Google Scholar]
  28. Stuenkel E. L., Hootman S. R. Secretagogue effects on intracellular calcium in pancreatic duct cells. Pflugers Arch. 1990 Aug;416(6):652–658. doi: 10.1007/BF00370610. [DOI] [PubMed] [Google Scholar]
  29. Stuenkel E. L., Machen T. E., Williams J. A. pH regulatory mechanisms in rat pancreatic ductal cells. Am J Physiol. 1988 Jun;254(6 Pt 1):G925–G930. doi: 10.1152/ajpgi.1988.254.6.G925. [DOI] [PubMed] [Google Scholar]
  30. Tamaoki T., Nomoto H., Takahashi I., Kato Y., Morimoto M., Tomita F. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase. Biochem Biophys Res Commun. 1986 Mar 13;135(2):397–402. doi: 10.1016/0006-291x(86)90008-2. [DOI] [PubMed] [Google Scholar]
  31. Vaandrager A. B., van den Berghe N., Bot A. G., de Jonge H. R. Phorbol esters stimulate and inhibit Cl- secretion by different mechanisms in a colonic cell line. Am J Physiol. 1992 Feb;262(2 Pt 1):G249–G256. doi: 10.1152/ajpgi.1992.262.2.G249. [DOI] [PubMed] [Google Scholar]
  32. Villanger O., Veel T., Raeder M. G. Secretin causes H+/HCO3- secretion from pig pancreatic ductules by vacuolar-type H(+)-adenosine triphosphatase. Gastroenterology. 1995 Mar;108(3):850–859. doi: 10.1016/0016-5085(95)90460-3. [DOI] [PubMed] [Google Scholar]
  33. Warhurst G., Higgs N. B., Tonge A., Turnberg L. A. Stimulatory and inhibitory actions of carbachol on chloride secretory responses in human colonic cell line T84. Am J Physiol. 1991 Aug;261(2 Pt 1):G220–G228. doi: 10.1152/ajpgi.1991.261.2.G220. [DOI] [PubMed] [Google Scholar]
  34. Zhao H., Star R. A., Muallem S. Membrane localization of H+ and HCO3- transporters in the rat pancreatic duct. J Gen Physiol. 1994 Jul;104(1):57–85. doi: 10.1085/jgp.104.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES