Role of H+ ions in volume and voltage of epithelial cell nuclei.
Condensation of chromatin depends upon the ion composition in the cell nucleus. We tested in isolated nuclei of Madin-Darby canine kidney cells the influence of various ions on nuclear volume (i. e. DNA packing) and intranuclear voltage. After isolation, nuclei were superfused with cytosolic solutions in which Na+, K+, Ca2+ and H+ ions were varied. With video-imaging and microelectrode techniques nuclear volume and intranuclear potential were measured in response to the various ions. In control cytosolic solution, isolated nuclei exhibited an intranuclear electrical potential of -6.5 +/- 0.5 mV (relative to a reference electrode in the cytosolic solution) corresponding to a nuclear volume of 250 +/- 10 fl (n = 104). Changing the Na+, K+ or free Ca2+ concentration in the superfusate in the physiological range resulted in minor changes of volume and intranuclear potential whereas pH altered both parameters dramatically. Nuclear swelling and intranuclear negative voltage increased with alkalinization and decreased when pH was reduced. An intact nuclear envelope was found to be no prerequisite for maintaining intranuclear negativity, indicating that the composition and functional state of nuclear chromatin rather than specific ion permeabilities of the nuclear envelope determine nuclear electrical potential. We present a model that explains nuclear volume and voltage on the basis of interaction between negatively charged DNA and positively charged histones of the nuclear chromatin.
Oberleithner
H
H
Department of Physiology, University of Würzburg, Germany.
Schuricht
B
B
Wünsch
S
S
Schneider
S
S
Püschel
B
B
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002412:Cations; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure*; D002843:Chromatin / Q000737:chemistry / Q000648:ultrastructure; D004247:DNA / Q000737:chemistry; D004285:Dogs; D004563:Electrochemistry; D004594:Electrophysiology; D004848:Epithelium / Q000648:ultrastructure; D006657:Histones / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000648:ultrastructure*; D009685:Nuclear Envelope / Q000502:physiology / Q000648:ultrastructure; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D011522:Protons; D012964:Sodium / Q000494:pharmacology
D002412:Cations; D002843:Chromatin; D006657:Histones; D011522:Protons; D011092:Polyethylene Glycols; D017830:Octoxynol; D004247:DNA; D012964:Sodium; D011188:Potassium; D002118:Calcium