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Artifacts caused by cell microinjection.

D S Miller, Y T Lau, S B Horowitz

    Proceedings of the National Academy of Sciences of the United States of America
    |March 1, 1984
    PubMed
    Summary
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    Microinjection into frog oocytes can cause artifactual solute leakage and diffusion gradients in aqueous solutions. Incubating cells in paraffin oil prevents these artifacts, ensuring accurate experimental results.

    Area of Science:

    • Cell Biology
    • Developmental Biology
    • Oocyte Physiology

    Background:

    • Microinjection is a common technique for introducing substances into cells.
    • Accurate interpretation of microinjection experiments relies on maintaining intracellular homeostasis.

    Purpose of the Study:

    • To investigate the effects of microinjection on the intracellular concentrations of sodium (Na), potassium (K), and water in Rana pipiens oocytes.
    • To assess the impact of microinjection on the distribution of injected radiolabeled sucrose.
    • To determine if microinjection artifacts can be avoided by altering the incubation medium.

    Main Methods:

    • Cryomicrodissection was used to measure Na, K, water, and radiolabeled sucrose.
    • Oocytes were microinjected and incubated in either Ringer's solution (aqueous) or paraffin oil.

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  • Intracellular solute and water distributions were compared between injected and uninjected oocytes.

    • Main Results:

    • Microinjection into Ringer's solution led to loss of K and sucrose, and gain of Na in the nucleus, ooplasm, and injection site, indicating solute leakage and diffusion gradients.
    • Despite leakage, oocytes underwent meiotic maturation when exposed to progesterone.
    • Microinjection and incubation in paraffin oil prevented changes in Na, K, and water concentrations, preserving normal solute distributions.

    Conclusions:

    • Microinjection in aqueous media can introduce artifacts, including solute loss and redistribution, which may not be apparent from overall cellular function.
    • Incubating microinjected oocytes in paraffin oil effectively prevents these artifacts, ensuring the integrity of intracellular solute and water concentrations.
    • These findings highlight the importance of considering the incubation medium in microinjection studies to avoid misinterpretation of experimental results.