Evaluating DNA damage response through immunofluorescence staining of primordial germ cells in Caenorhabditis elegans L1 larva

Summary C. elegans L1 larvae have two well-defined primordial germ cells embedded in a niche comprising two somatic gonad precursor cells. Thus, C. elegans provides an ideal model for studying intercellular signaling in response to DNA damage. However, existing staining protocols are focused on worms in later developmental stages and are not optimized for the L1 larvae. Here, we present a revised protocol for assessing the DNA damage response utilizing immunofluorescence staining specifically in C. elegans L1 larva. For complete details on the use and execution of this protocol, please refer to Ou et al. (2019).
Graphical abstract
Highlights • Primordial germlines in C. elegans L1 larva as ideal stem cell-niche interaction models • Evaluating the intercellular DDR within microenvironment by PGC/SGP immunostaining • Optimized staining and quantification protocols for C. elegans L1 larva
C. elegans L1 larvae have two well-defined primordial germ cells embedded in a niche comprising two somatic gonad precursor cells. Thus, C. elegans provides an ideal model for studying intercellular signaling in response to DNA damage. However, existing staining protocols are focused on worms in later developmental stages and are not optimized for the L1 larvae. Here, we present a revised protocol for assessing the DNA damage response utilizing immunofluorescence staining specifically in C. elegans L1 larva.