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Related Experiment Videos

Bacterial mesosomes. Real structures or artifacts?

M T Silva, J C Sousa, J J Polónia

    Biochimica Et Biophysica Acta
    |August 4, 1976
    PubMed
    Summary
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    Ultrastructural studies reveal that large bacterial mesosomes are artifacts of osmium tetroxide (OSO4) fixation, not natural structures. Alternative fixation methods suggest a continuous cytoplasmic membrane in gram-positive bacteria.

    Area of Science:

    • Microbiology
    • Cell Biology
    • Microscopy

    Background:

    • Mesosomes have been historically described as invaginations of the cytoplasmic membrane in gram-positive bacteria.
    • The Ryter-Kellenberger procedure, a common fixation method, involves an initial prefixation step with osmium tetroxide (OSO4).

    Purpose of the Study:

    • To investigate the role of OSO4 fixation conditions in the observed ultrastructure of mesosomes in gram-positive bacteria.
    • To determine if mesosomes are true cellular structures or artifacts of specific fixation techniques.

    Main Methods:

    • Comparative ultrastructural analysis of Bacillus cereus and Streptococcus faecalis using various fixation protocols.
    • Kinetic study of membrane morphological changes during OSO4 prefixation.
    • Assessment of OSO4's effect on protoplast integrity and intracellular potassium (K+) leakage.

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    Main Results:

    • Large, complex mesosomes were observed exclusively when using 0.1% OSO4 prefixation, characteristic of the Ryter-Kellenberger method.
    • OSO4 prefixation rapidly damaged bacterial membranes, causing protoplast lysis and K+ leakage, suggesting it induces artifactual structures.
    • Fixation with glutaraldehyde or higher concentrations of OSO4, or with uranyl acetate, resulted in minimal or no large mesosomes, showing only simple membrane invaginations or none at all.

    Conclusions:

    • The formation of large mesosomes is an artifact induced by the membrane-damaging effects of 0.1% OSO4 prefixation.
    • The study casts doubt on the existence of both large and small mesosomes as genuine structures in gram-positive bacteria.
    • It is proposed that gram-positive bacteria possess a continuous cytoplasmic membrane without complex infoldings, challenging established models of bacterial membrane organization.