Increase in cytosolic Ca2+ regulates exocytosis and Cl- conductance in HT29 cells.
Increases of cytosolic Ca2+, as occur with agonists such as ATP, neurotensin (NT), hypotonic cell swelling and ionomycin, enhance the membrane conductance (GM) and hence the input conductance (GI) of HT29 cells. In the present study we have examined whether these increases in GM are paralleled by exocytosis. To this end the membrane capacitance (CM) of HT29 cells was measured by patch clamp techniques. Two methods to monitor CM were used: a direct method (DM) and a phase tracking method (PTM). With the DM the following results were obtained. NT (10(-8) mol/l, n = 9) increased GM and CM significantly from 2.4 +/- 0.3 nS and 23.5 +/- 3 pF to 32 +/- 8 nS and 27.3 +/- 3.1 pF respectively. ATP (10(-4) mol/l, n = 29) had a very similar effect. GM and CM were increased from 5.7 +/- 1 nS and 36 +/- 4.4 pF to 111 +/- 21 nS and 44 +/- 5.4 pF respectively. Hypotonic cell swelling (160 mosmol/l, n = 18) had a comparable effect: GM and CM were increased from 4.9 +/- 1 nS and 30 +/- 4.1 pF to 46 +/- 10 nS and 37 +/- 4.9 pF respectively. Ionomycin (10(-7) mol/l, n = 4) gave similar results. With the PTM it was possible to monitor the rapid changes in GM and CM, as they were induced by ATP (n = 42) and NT (n = 29), with high time resolution.(ABSTRACT TRUNCATED AT 250 WORDS)
Greger
R
R
Physiologisches Institut, Albert-Ludwigs-Universität, Freiburg, Germany.
Allert
N
N
Fröbe
U
U
Normann
C
C
D000255:Adenosine Triphosphate / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D018118:Chloride Channels / Q000187:drug effects / Q000378:metabolism*; D003110:Colonic Neoplasms / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D005089:Exocytosis / Q000502:physiology*; D006801:Humans; D007038:Hypotonic Solutions; D015759:Ionomycin / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D009496:Neurotensin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D018118:Chloride Channels; D007038:Hypotonic Solutions; D009496:Neurotensin; D015759:Ionomycin; D000255:Adenosine Triphosphate; D002118:Calcium