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PC12 variants deficient in catecholamine transport.

C M Bitler, M B Zhang, B D Howard

    Journal of Neurochemistry
    |October 1, 1986
    PubMed
    Summary
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    Researchers created PC12 cell variants lacking dopamine transporter function. MPTP-resistant variants showed multiple deficiencies, including impaired dopamine storage and nerve growth factor response, indicating broader impacts beyond transport.

    Area of Science:

    • Neuroscience
    • Cell Biology
    • Pharmacology

    Background:

    • PC12 cells are a model system for studying neuronal function.
    • Transporter-mediated uptake is crucial for neurotransmitter regulation.
    • N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a neurotoxin that targets dopamine transporters.

    Purpose of the Study:

    • To isolate and characterize PC12 cell variants with defects in dopamine transporter-mediated uptake.
    • To investigate the functional consequences of impaired dopamine transport in these variants.
    • To identify molecular differences between wild-type and variant PC12 cells.

    Main Methods:

    • Isolation of PC12 cell variants resistant to guanethidine or MPTP.
    • Measurement of dopamine uptake kinetics (Vmax, Km).

    Related Experiment Videos

  • Assessment of cellular responses to nerve growth factor.
  • Protein labeling with [3H]xylamine and Northern blot analysis for mRNA levels.

    • Main Results:

    • Guanethidine-resistant variants showed decreased Vmax for dopamine uptake.
    • MPTP-resistant variants completely lacked dopamine uptake capacity.
    • MPTP-resistant cells exhibited deficiencies in dopamine/acetylcholine storage and nerve growth factor response.
    • [3H]xylamine labeling revealed reduced protein labeling in variants, with three proteins missing in MPTP-resistant cells.
    • Northern blots indicated reduced mRNA levels for specific species in MPTP-resistant cells.

    Conclusions:

    • Dopamine transporter deficiency can be modeled using PC12 cell variants.
    • MPTP-resistant variants display pleiotropic defects beyond dopamine transport.
    • These variants provide tools to study dopamine transporter function and its cellular impact.