Elimination of mitochondrial elements and improved viability in hybrid cells

Abstract

Experiments were carried out to determine whether the mitochondria-specific dye rhodamine-6G (R6G) can affect transmission of cytoplasmic determinants in mammalian cells. When one parental cell type was treated with R6G prior to fusion with an untreated partner, the subsequent hybridization frequencies in both intra- and interspecific crosses were not adversely affected, even though R6G was extremely toxic to the parental cells. In addition, cells lethally treated with R6G could be rescued by fusion with cytoplasm alone from untreated cells. When chloramphenicol (CAP) resistant cells were used as the R6G-treated parent, the expression of CAP resistance in hybrids and cybrids was greatly reduced. Thus R6G can be used to control the input of cytoplasmic determinants into fused cells. In the interspecific (Chinese hamster x mouse) crosses, it was also seen that the majority of hybrids which had not been R6G pretreated grew poorly or degenerated after a short time. In contrast, nearly all hybrids in crosses where the hamster parent was R6G pretreated grew vigorously. The concomitant elimination of inviability and loss of mitochondrial determinants in R6G-pretreated hybrids suggests that interactions involving mitochondrial gene products or components can influence growth characteristics in interspecific somatic cell hybrids.