Studies on human porin XXI: gadolinium opens Up cell membrane standing porin channels making way for the osmolytes chloride or taurine-A putative approach to activate the alternate chloride channel in cystic fibrosis.
We recently proposed that cell-membrane-integrated vertebrate porin/voltage-dependent anion-selective channel (VDAC) forms part of the outwardly rectifying chloride channel (ORCC) complex that may be involved in volume regulation. The results we present here support this thesis. According to light scattering measurements micromolar concentrations of Gd(3+) induce cell swelling of human healthy and cystic fibrosis (CF) B-lymphocyte cell lines in isotonic Ringer solution. In high-potassium Ringer solution additional swelling is observed. Gd(3+) induces excessive cell swelling of cell lines in hypotonic Ringer solutions, containing 70 mM NaCl or 135 mM taurine, respectively. The gadolinium effect is lost when NaCl is replaced by Na-gluconate. Using video camera monitoring we show that HeLa cells also swell in micromolar concentrations of Gd(3+) in isotonic taurine Ringer solution. The dose-dependent effect of the agonist was always blocked by extracellular application of anti-human type-1 porin antibodies. Together with data on a decreasing effect of micromolar amounts of gadolinium on the voltage dependence of reconstituted human porin the results prove the involvement of porin channels in the swelling behavior in different cell lines. As a mechanism we propose that ionic gadolinium opens up plasmalemma-integrated porin channels, chloride or taurine then following their concentration gradients into the cells. Furthermore, our data argue for a single pathway for inorganic and organic osmolytes during regulatory volume decrease after cell swelling. There is indirect evidence that porin forms part of the cystic fibrosis relevant ORCC channel. Gadolinium thus may work to open the alternate chloride channel in CF.
Thinnes
F P
FP
Abteilung Immunchemie, Max-Planck-Institut für Experimentelle Medizin, Hermann-Rein-Strasse 3, Göttingen, D-37075, Germany. Thinnes@EXMDI1.MPIEM.GWDG.DE
Hellmann
K P
KP
Hellmann
T
T
Merker
R
R
Schwarzer
C
C
Walter
G
G
Götz
H
H
Hilschmann
N
N
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D048429:Cell Size / Q000187:drug effects; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology; D002712:Chlorides / Q000493:pharmacokinetics; D003550:Cystic Fibrosis / Q000503:physiopathology; D004305:Dose-Response Relationship, Drug; D005682:Gadolinium / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D007038:Hypotonic Solutions / Q000494:pharmacology; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology; D007552:Isotonic Solutions / Q000494:pharmacology; D008565:Membrane Proteins / Q000187:drug effects / Q000502:physiology; D018272:Porins / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D000077331:Ringer's Solution; D013654:Taurine / Q000493:pharmacokinetics; D013997:Time Factors
D000911:Antibodies, Monoclonal; D018118:Chloride Channels; D002712:Chlorides; D007038:Hypotonic Solutions; D007473:Ion Channels; D007552:Isotonic Solutions; D008565:Membrane Proteins; D018272:Porins; D013654:Taurine; D000077331:Ringer's Solution; D005682:Gadolinium