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Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy
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Canonical insulin-like receptor DAF-2 signaling-independent patterning and role for FoxO/DAF- 16 in early embryos

Michael S Mauro, Jessica M Peiser, Sophia L Martin

    Biorxiv : the Preprint Server for Biology
    |August 20, 2026
    PubMed

    Insights

    Forkhead box O (FoxO) transcription factors, like DAF-16 in C. elegans, are crucial for development. This study reveals DAF-16

    Area of Science:

    • Developmental Biology
    • Genetics
    • Cell Biology

    Background:

    • Forkhead box O (FoxO) transcription factors, including DAF-16 in *C. elegans*, are vital for conserved processes like metabolism, aging, and development.
    • Insulin receptor signaling (DAF-2 in *C. elegans*) typically inhibits FoxO/DAF-16, but other regulatory pathways exist.
    • Severe loss-of-function mutations in *daf-2* and *daf-16* together cause high embryonic lethality in *C. elegans*, a phenomenon not understood.

    Purpose of the Study:

    • To investigate the role and localization of DAF-16 during early *C. elegans* embryogenesis.
    • To elucidate the cause of synthetic lethality observed in *daf-2*; *daf-16* double mutants.
    • To identify novel regulatory pathways and functions of DAF-16 in embryonic development.

    Main Methods:

    • Utilized genetics and quantitative imaging techniques in *C. elegans* embryos.
    • Performed lineage tracing to track cell fate and DAF-16 localization.
    • Analyzed mitotic chromosome segregation and morphogenesis in mutant embryos.

    Main Results:

    • DAF-16 exhibited uniform low-level nuclear localization before the 8-cell stage.
    • From the 8-cell to 64-cell stages, DAF-16 became enriched in germ lineage cell nuclei.
    • This germ lineage-specific DAF-16 patterning was dependent on germ fate determinants and PTEN/DAF-18, but independent of DAF-2 activity and maternal age.
    • Severe *daf-16; daf-2* double mutants displayed significant defects in morphogenesis and chromosome segregation from the 1-cell stage.

    Conclusions:

    • Identified a novel germ lineage-specific localization pattern for DAF-16 in early *C. elegans* embryos.
    • Demonstrated a DAF-2 signaling-independent function for DAF-16 critical for early embryogenesis, including morphogenesis and chromosome segregation.
    • The findings shed light on the complex regulation and essential roles of DAF-16 beyond canonical insulin signaling pathways.

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