Electrical measurements of complement-mediated membrane damage in cultured nerve and muscle cells.
Cultured neuroblastoma-glioma hybrid cells and primary mouse and rat muscle cells were studied by using intracellular microelectrodes to monitor membrane electrical potential and resistance changes during complement-mediated lysis. The cell membrane was TNP modified under mild conditions and subsequently coated with rabbit IgG anti-TNP, with no electrical changes observed. However, upon addition of guinea pig C the membrane potential dropped from approximately -50 mv to less than -12 mv within a few minutes, with parallel decreases in electrical resistance. Ten to 60 min after these electrical changes the cells became stainable with trypan blue. No electrical changes or trypan blue staining was observed in the absence of antibody or with heat-inactivated C. With more dilute C so that only a fraction of cells became trypan blue positive, all cells nevertheless showed the membrane electrical changes; surviving cells recovered their original membrane properties within 1 hr. Thus the C-mediated damage to the membrane measured electrically is not in itself sufficient to ensure the subsequent death of the cell. The early electrical changes observed appear to be comparable to increases in 86Rb efflux measured by others.
D000818:Animals; D000906:Antibodies; D002462:Cell Membrane / Q000276:immunology; D002470:Cell Survival; D002478:Cells, Cultured; D003165:Complement System Proteins / Q000378:metabolism*; D004558:Electric Stimulation; D005910:Glioma / Q000276:immunology; D006168:Guinea Pigs; D008564:Membrane Potentials; D051379:Mice; D009132:Muscles / Q000378:metabolism*; D009447:Neuroblastoma / Q000276:immunology; D009474:Neurons / Q000378:metabolism*; D011817:Rabbits; D051381:Rats
D000906:Antibodies; D003165:Complement System Proteins