Cybrid models of mtDNA disease and transmission, from cells to mice.

Oxidative phosphorylation (OXPHOS) is the only mammalian biochemical pathway dependent on the coordinated assembly of protein subunits encoded by both nuclear and mitochondrial DNA (mtDNA) genes. Cytoplasmic hybrid cells, cybrids, are created by introducing mtDNAs of interest into cells depleted of endogenous mtDNAs, and have been a central tool in unraveling effects of disease-linked mtDNA mutations. In this way, the nuclear genetic complement is held constant so that observed effects on OXPHOS can be linked to the introduced mtDNA. Cybrid studies have confirmed such linkage for many defined, disease-associated mutations. In general, a threshold principle is evident where OXPHOS defects are expressed when the proportion of mutant mtDNA in a heteroplasmic cell is high. Cybrids have also been used where mtDNA mutations are not known, but are suspected, and have produced some support for mtDNA involvement in more common neurodegenerative diseases. Mouse modeling of mtDNA transmission and disease has recently taken advantage of cybrid approaches. By using cultured cells as intermediate carriers of mtDNAs, ES cell cybrids have been produced in several laboratories by pretreatment of the cells with rhodamine 6G before cytoplast fusion. Both homoplasmic and heteroplasmic mice have been produced, allowing modeling of mtDNA transmission through the mouse germ line. We also briefly review and compare other transgenic approaches to modeling mtDNA dynamics, including mitochondrial injection into oocytes or zygotes, and embryonic karyoplast transfer. When breakthrough technology for mtDNA transformation arrives, cybrids will remain valuable for allowing exchange of engineered mtDNAs between cells.