Microelectrode measurements of the transmembrane potential in baby hamster kidney, (Chinese hamster ovary), NG108-15 neuroblastoma and Swiss 3T3 cells at 37.0 or 43.0 degrees C.
Hyperthermia affects the physical state and function of the plasma membrane. This could alter the transmembrane potential (Vm) and associated functions in a manner that promotes cell killing. Previous investigations have reported differing results of the effect of heat on Vm, possibly due to artifacts associated with the methods employed to measure Vm indirectly. One such artifact is a membrane depolarization induced by cationic probes, as demonstrated in this paper. In this study, glass microelectrodes were used to avoid these artifacts and to make direct electrical measurements of Vm. Following 25 min-30 min at 43.0 degrees C, The mean Vm of Chinese hamster ovary (CHO) cells, within clusters of six or more cells, decreased from -16 +/- 5 to -38 +/- 6 mV, and remained at these levels during incubation times up to 3 h. All CHO cells resumed a normal Vm within 4.5 h after returning to 37.0 degrees C, regardless of the time of exposure at 43.0 degrees C (0.5 to 3.0 h, with survival levels of 0.7 and 0.001, respectively). The membrane hyperpolarization decreased with cell to cell contact to where isolated cells exhibited no hyperpolarization. CHO cultures with different cell densities (number of cells per cm2), and thus differing degrees of cell to cell contact, were heated and then subjected to the colony formation assay. The degree of cell to cell contact at the time of heating had no effect on survival. Hence, the heat-induced, cell contact dependent hypolarization of CHO cell membranes was unrelated to clonogenic survival.(ABSTRACT TRUNCATED AT 250 WORDS)
Borrelli
M J
MJ
Department of Radiation Oncology, William Beaumont Hospital, Royal Oak, MI 48073.
Rausch
C M
CM
D016475:3T3 Cells; D000818:Animals; D016466:CHO Cells; D002232:Carbocyanines / Q000494:pharmacology; D002448:Cell Adhesion; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D005456:Fluorescent Dyes / Q000494:pharmacology; D006358:Hot Temperature; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008839:Microelectrodes; D014407:Tumor Cells, Cultured
D002232:Carbocyanines; D005456:Fluorescent Dyes; C018528:3,3'-dipentyl-2,2'-oxacarbocyanine