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Regulation of chloride secretion in HT29 cells studied by X-ray microanalysis
Uppsala universitet, Institutionen för anatomi.
1997 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Several aspects of Cl- secretion in epithelial cells were investigated. Methods to prepare cultured cellsfor X-ray microanalysis, in order to study volume-regulating chloride transport were evaluated. Inaddition, α2A-adrenergic, muscarinic cholinergic, and purinergic Cl- efflux as well as Cl- effluxregulated by G-protein coupled receptors in HT29 cells was investigated. The purpose of the studywas to describe the regulation of Cl- transport in intestinal epithelial cells.

Exposure of HT29 cells to a hypotonic solution resulted in regulatory volume decrease (RVD)accompanied by loss of K and Cl from the cells, whereas exposure to hypertonic solutions resultedin regulatory volume increase (RVI) coupled to gain of Na and Cl. Chloride channel blockers, suchas NPPB and DPC, blocked RVD, accompanied by a decrease in intracellular Na, Cl and Kconcentration, suggesting that chloride channels are important in volume regulating ion transport.

Stimulation with UTP or ATP mainly induced loss of Cl and Na from the cells, concurrent with an increase in Ca. The ion fluxes induced by UTP could be blocked by NPPB and alloxan (aninhibitor of adenylate cyclase), indicating that UTP-mediated Cl- secretion may occur through bothcAMP- and Ca2+-dependent pathways.

The agonists of muscarinic receptors, carbachol and acetylcholine, caused a decrease inintracellular Cl and K concentration. However, P-F-HHSiD, an antagonist of muscarinic 3 receptors,inhibited secretion induced by carbachol or acetylcholine, suggesting that the ion secretion is mediatedby muscarinic 3 receptors. Moreover, the ion secretion could be inhibited by U-73122, an inhibitorof phospholipase C (PLC), indicating that secretion may be regulated by the PLC/Ca2+ dependent pathway.

Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating polypeptide(PACAP) elicited Cl- and K+ efflux, whereas UK14,304 and somatostatin-14 were able to reverse VIPor PACAP induced secretion, giving rise to an increase in the content of intracellular Na and Cl. Theantisecretory effect of UK14,304 and somatostatin-14 is probably involved in the inhibition of bothadenylate cyclase and PLC pathways by relevant receptors coupled to Gi Use of U-73122 and alloxantogether inhibited PACAP-induced Cl- efflux, indicatingthat Cl- secretion may not only be mediatedby cAMP activated Cl- channels, but also by Ca2+ activated Cl- channels.

sted, utgiver, år, opplag, sider
Uppsala: Acta Universitatis Upsaliensis , 1997. , s. 35
Serie
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 0282-7476 ; 732
Emneord [en]
Anatomy, epithelium, chloride channel, HT29 cells, ion transport, volume, X-ray microanalysis, G-protein coupled receptors, α2-adrenoceptors, purinoceptors, muscarinic receptors
Emneord [sv]
Anatomi
HSV kategori
Forskningsprogram
anatomi
Identifikatorer
URN: urn:nbn:se:uu:diva-1220ISBN: 91-554-4103-3 (tryckt)OAI: oai:DiVA.org:uu-1220DiVA, id: diva2:160778
Disputas
1997-12-19, Seminar room C3, Uppsala Biomedical Center, Uppsala, 09:15
Tilgjengelig fra: 1997-11-28 Laget: 1997-11-28 Sist oppdatert: 2018-01-13bibliografisk kontrollert

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