Regulation of high-conductance anion channels by G proteins and 5-HT(1A) receptors in CHO cells

Allen W. Mangel, John R. Raymond, J. Gregory Fitz

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

This study addresses the mechanisms responsible for regulation of high- conductance anion channels by GTP binding proteins in Chinese hamster ovary (CHO) cells. Single-channel currents were measured in inside-out membrane patches using patch-clamp techniques. Anion-selective channels with a unitary conductance of 381 ± 8 pS activated spontaneously in 48% of excised patches. In patches with no spontaneous channel activity, addition of GppNHp, a nonhydrolyzable analogue of GTP, activated channels in 8 of 12 studies, and in patches with spontaneous channel activity, GppNHp increased open probability in 4 of 4 experiments. In contrast, GDPβS, a nonhydrolyzable GDP analogue, inhibited both spontaneous and GppNHp-induced channel activity. In patches without spontaneous channel activity, addition of cholera toxin activated channels in five of eight studies. Interestingly, pertussis toxin had a similar effect, activating channels in five of seven previously quiescent patches. To further evaluate the possible role of inhibitory G proteins in channel regulation, activity was measured in cell-attached patches in cells transfected with the serotonin 5-HT(1A) receptor, which is coupled to effector mechanisms through a pertussis toxin-sensitive G protein. Stimulation of 5-HT(1A)-transfected cells with the receptor agonist (±)-8- hydroxy-2-(di-n-propylamino)tetralin caused a transient decrease in open probability in either standard or high-potassium solutions. In aggregate, these findings suggest that both cholera and pertussis toxin-sensitive G proteins contribute to regulation of high-conductance anion channels in CHO cells.

Original languageEnglish (US)
Pages (from-to)F490-F495
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume264
Issue number3 33-3
StatePublished - Apr 16 1993

Keywords

  • chloride ion channels
  • cholera toxin
  • patch clamp
  • pertussis toxin
  • serotonin 5-HT(1A) receptors

ASJC Scopus subject areas

  • Physiology

Fingerprint Dive into the research topics of 'Regulation of high-conductance anion channels by G proteins and 5-HT(1A) receptors in CHO cells'. Together they form a unique fingerprint.

  • Cite this