Lai C N, Gallick G E, Arlinghaus R B, Becker F F
J Cell Physiol. 1984 Oct;121(1):139-42. doi: 10.1002/jcp.1041210117.
Normal rat kidney cells (NRK) infected with the temperature-sensitive (ts) transformation mutant of Moloney murine sarcoma virus yielded a clone of cells, 6m2, that exhibited a transformed morphology at 33 degrees C and a normal morphology at 39 degrees C. Transmembrane potential (Em) was measured fluorometrically using a cyanine dye diS-C3-(5). Fluorescence was inversely correlated with Em. Cells at 33 degrees C had lower Em. Em changes were recorded within 15 minutes of temperature shift from 33 degrees C to 39 degrees C in both directions, increasing in the 33 degrees C to 39 degrees C direction and decreasing in the 39 degrees C to 33 degrees C direction. Uninfected NRK cells when shifted under the same condition exhibited small fluorescence changes in the 33 degrees C to 39 degrees C direction. Shifting from 39 degrees C to 33 degrees C resulted in Em changes similar to those in 6m2 cells. Also studied was a cell line infected with a spontaneous revertant of the ts mutant, designated 54-5A4; it was transformed at both temperatures. Shifting from 33 degrees C to 39 degrees C in both directions yielded small changes. Transmembrane potential changes in 6m2 cells precede other transformation-specific changes that occur after a temperature shift.
感染莫洛尼鼠肉瘤病毒温度敏感(ts)转化突变体的正常大鼠肾细胞(NRK)产生了一个细胞克隆,即6m2,该克隆在33℃时呈现转化形态,在39℃时呈现正常形态。使用花青染料diS-C3-(5)通过荧光法测量跨膜电位(Em)。荧光与Em呈负相关。33℃的细胞Em较低。在温度从33℃双向转移到39℃的15分钟内记录到Em变化,在从33℃到39℃的方向上增加,在从39℃到33℃的方向上减少。未感染的NRK细胞在相同条件下转移时,在从33℃到39℃的方向上表现出微小的荧光变化。从39℃转移到33℃导致的Em变化与6m2细胞中的变化相似。还研究了一种感染ts突变体自发回复体的细胞系,命名为54-5A4;它在两个温度下均被转化。双向从33℃转移到39℃产生微小变化。6m2细胞中的跨膜电位变化先于温度转移后发生的其他转化特异性变化。