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Fusion of mycoplasmas: the formation of cell hybrids
M Tarshis1, M Salman, S Rottem
1Department of Membrane and Ultrastructure Research, Hebrew University, Hadassah Medical School, Jerusalem, Israel.
Abstract:
Poly(ethylene glycol) (PEG 8000) can induce cell-cell fusion of Mycoplasma capricolum cells, and it can promote the formation of intergeneric hybrids of various Mycoplasma, Acholeplasma and Spiroplasma species. The extent of fusion was quantitatively evaluated by following the dequenching of octadecylrhodamine fluorescent label incorporated into donor cell membranes after their incubation with recipient cells. The results of dequenching experiments were confirmed by electron microscopy, as well as by angle light-scattering measurements. Fusion appeared to require the presence of Mg2+, but was completely inhibited by either 0.1% glutaraldehyde or 100 microM chlorpromazine, and was partially suppressed by proteolytic enzymes, carbonyl cyanide-m-chlorophenylhydrazone, or thiol reagents.
Insights
Poly(ethylene glycol) (PEG 8000) effectively induces cell fusion in Mycoplasma species, promoting intergeneric hybrid formation. This process requires magnesium ions and is inhibited by specific chemicals and enzymes.
Area of Science:
- Microbiology
- Cell Biology
- Biotechnology
Background:
- Mycoplasma species are bacteria lacking cell walls, posing challenges for genetic manipulation.
- Cell-cell fusion is a key technique for creating hybrid organisms and studying cellular interactions.
Purpose of the Study:
- To investigate the efficacy of Poly(ethylene glycol) (PEG 8000) in inducing cell-cell fusion among Mycoplasma, Acholeplasma, and Spiroplasma species.
- To quantitatively assess the extent of fusion and identify factors influencing the process.
Main Methods:
- Utilized octadecylrhodamine fluorescent labeling to quantify cell fusion by monitoring dequenching.
- Confirmed fusion extent using electron microscopy and angle light-scattering measurements.
- Investigated the effects of various chemical and enzymatic treatments on fusion.
Main Results:
- Poly(ethylene glycol) (PEG 8000) successfully induced cell-cell fusion in Mycoplasma capricolum and promoted intergeneric hybrid formation.
- Fusion efficiency was quantitatively measured using fluorescence dequenching, corroborated by microscopy and light scattering.
- Fusion was dependent on Mg2+ and inhibited by glutaraldehyde, chlorpromazine, proteolytic enzymes, and thiol reagents.
Conclusions:
- Poly(ethylene glycol) (PEG 8000) is a potent agent for inducing Mycoplasma cell fusion and generating intergeneric hybrids.
- Understanding fusion requirements and inhibitors provides insights for optimizing cell fusion techniques in mycoplasmal research.