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Isolation, Fixation, and Immunofluorescence Imaging of Mouse Adrenal Glands
08:37

Isolation, Fixation, and Immunofluorescence Imaging of Mouse Adrenal Glands

Published on: October 2, 2018

Mutations in cyclic AMP-dependent protein kinase and corticotropin (ACTH)-sensitive adenylate cyclase affect adrenal

P A Rae, N S Gutmann, J Tsao

    Proceedings of the National Academy of Sciences of the United States of America
    |April 1, 1979
    PubMed

    Insights

    Mutant adrenocortical cells reveal key signaling pathways. Cyclic AMP-dependent protein kinase activity, not just adenylate cyclase, is crucial for corticotropin-stimulated steroidogenesis.

    Area of Science:

    • Endocrinology
    • Molecular Biology
    • Cell Signaling

    Background:

    • Adrenocortical tumor cells (Y1) are a model for studying steroidogenesis.
    • Corticotropin stimulates steroid production via intracellular signaling cascades.

    Purpose of the Study:

    • To investigate the roles of adenylate cyclase and cyclic AMP-dependent protein kinase in corticotropin-stimulated steroidogenesis.
    • To elucidate the signaling pathway components essential for hormone action.

    Main Methods:

    • Isolation and characterization of two distinct mutant Y1 cell lines: Y1(Kin) with altered protein kinase activity and Y1(Cyc) with diminished adenylate cyclase activity.
    • Assessment of steroidogenic responses to corticotropin and cyclic nucleotides in mutant cell lines.

    Main Results:

    • Y1(Kin) mutants showed steroidogenic responses that paralleled cyclic AMP-dependent protein kinase activity.
    • Y1(Cyc) mutants displayed reduced steroidogenesis in response to corticotropin but maintained responsiveness to cyclic nucleotides.
    • These findings differentiate the roles of adenylate cyclase and protein kinase in the pathway.

    Conclusions:

    • Adenylate cyclase and cyclic AMP-dependent protein kinase are indispensable components of the corticotropin-stimulated steroidogenic pathway.
    • The data suggest a sequential or parallel involvement of these enzymes in mediating hormone action.
    • This study clarifies the molecular requirements for adrenocortical cell stimulation.

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