Harnessing Microbial Tools: Escherichia coli as a Vehicle for Neuropeptide Functional Analysis in Caenorhabditis elegans

Animals respond to changes in their environment and internal states via neuromodulation. Neuropeptides modulate neural circuits with flexibility because one gene can produce either multiple copies of the same neuropeptide or different neuropeptides. However, with this architectural complexity, the function of discrete and active neuropeptides is muddled. Here, we design a genetic tool that facilitates functional analysis of individual peptides. We engineered Escherichia coli bacteria to express active peptides, fed loss-of-function Caenorhabditis elegans, and rescued the activity of genes with varying lengths and functions: pdf-1, flp-3, ins-6, and ins-22. Some peptides were functionally redundant, while others exhibit unique and previously uncharacterized functions. We postulate our rescue-by-feeding approach can elucidate the functional landscape of neuropeptides, identifying the circuits and complex peptidergic pathways that regulate different behavioral and physiological processes. Article summaryStudying individual neuropeptides opens new avenues for exploring neuromodulation at a finer resolution. The researchers developed a method to create DNA vectors that encode an endogenous peptide sequence flanked by sequences containing dibasic endopeptidase cleavage sites in Caenorhabditis elegans. The researchers transformed these vectors into bacteria and fed them to C. elegans, which restored wildtype behavior in neuropeptide loss-of-function mutants. The researchers also discovered that neuropeptides from the same gene perform distinct functions, a research area more ready to explore using the presented technology. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=176 HEIGHT=200 SRC="FIGDIR/small/641308v2_ufig1.gif" ALT="Figure 1"> View larger version (58K): org.highwire.dtl.DTLVardef@9153a1org.highwire.dtl.DTLVardef@109788dorg.highwire.dtl.DTLVardef@c97289org.highwire.dtl.DTLVardef@17eb30a_HPS_FORMAT_FIGEXP M_FIG Created in BioRender. DiLoreto, E. (2025) https://BioRender.com/ypbrtlk C_FIG