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Related Experiment Videos

Control of elastin synthesis

W Burnett, K Yoon, A Finnigan-Bunick

    The Journal of Investigative Dermatology
    |July 1, 1982
    PubMed
    Summary

    Researchers developed a probe to measure elastin messenger RNA (mRNA) in chick embryos. They found that elastin mRNA levels increase during development and in response to hydrocortisone, directly impacting elastin synthesis.

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    Area of Science:

    • Molecular Biology
    • Developmental Biology
    • Biochemistry

    Background:

    • Elastin is a crucial protein for tissue elasticity, particularly in blood vessels.
    • Understanding the regulation of elastin synthesis during embryonic development is essential for cardiovascular health.

    Purpose of the Study:

    • To isolate and characterize elastin messenger RNA (mRNA) from chick embryo aortas.
    • To develop a method for quantifying elastin mRNA levels and their correlation with elastin synthesis.
    • To investigate the effect of hydrocortisone on elastin mRNA abundance and protein synthesis.

    Main Methods:

    • Synthesis of complementary DNA (cDNA) from mRNA using reverse transcriptase and nuclease treatment.
    • Cloning of elastin cDNA into plasmid pBR322 and transformation of E. coli.
    • Selection of elastin cDNA clones using in situ hybridization and hybrid-selected translation.
    • Sequencing of the cDNA insert and its use as a probe for blot hybridization.
    • Quantification of elastin mRNA levels in developing chick embryos and after hydrocortisone administration.

    Main Results:

    • A recombinant plasmid, pWB1, containing a 230 bp elastin cDNA insert was isolated.
    • This clone's mRNA stimulated the synthesis of a 72,000 molecular weight protein immunoprecipitable as tropoelastin.
    • Blot hybridization identified a 3.5 kb elastin mRNA species that increased significantly between embryonic days 7 and 14.
    • Elastin mRNA levels and synthesis rates paralleled each other during development.
    • Hydrocortisone administration increased both tropoelastin synthesis and elastin mRNA levels.

    Conclusions:

    • Elastin synthesis during chick embryo development is regulated by the abundance of elastin mRNA.
    • Hydrocortisone treatment influences elastin synthesis by modulating elastin mRNA content.
    • The developed cDNA probe is a valuable tool for studying elastin gene expression.

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