Rohlfs, M. | Search bioRxiv

bioRxiv2025

Ecological context shapes microbial contributions to nutrition and development in Drosophila melanogaster

Microbial symbionts play a critical role in shaping insect development by supplying essential nutrients, influencing host phenotype, and mediating context-dependent mutualistic outcomes. In the fruit fly Drosophila melanogaster, larvae develop in ephemeral breeding substrates such as decaying fruits, where they acquire maternally transmitted bacterial and fungal symbionts. These substrates vary in their structural and chemical properties, potentially altering the composition and function of associated microbial communities. Here, we combine semi-natural microcosm experiments with compound-specific isotope analysis of essential amino acids to assess how variation in fruit substrate and symbiont origin affect larval nutrition, development, and adult phenotypes. We find that microbial community composition and environmental context jointly shape amino acid flow, with larvae deriving variable proportions of their nutrition from plant, bacterial, and fungal sources. These differences translate into distinct developmental outcomes, including shifts in larval development time - either delayed or accelerated, depending on the symbiont-substrate combination - and significant variation in adult body weight. Our findings introduce symbiont-environment interactions as a framework for understanding how ecological context shapes nutritional mutualisms, showing that environmental heterogeneity can alter microbial composition, nutrient sources, and larval performance. Together, these results reveal how ecological context modulates mutualistic dependence and underscores the context sensitivity of host-microbe associations.

ecology↗

bioRxiv2025

Mutations in VPS18 lead to a neutrophil maturation defect associated with disturbed vesicle homeostasis

Neutrophils, the first cells to arrive at the site of inflammation, are rather short-lived cells and thus have to be constantly replenished. During neutrophil development, vesicle dynamics need to be fine-tuned and impaired vesicle trafficking has been linked to failure in neutrophil maturation. Here, we characterized the role of VPS18 as a central core component of CORVET & HOPS tethering complexes for neutrophil development. Using CRISPR/Cas9-engineered Hoxb8 cells with heterozygous mutations in Vps18, we found that VPS18 deficiency interfered with neutrophil development due to tethering complex instability. As a result, vesicle dynamics were impaired with a strong increase in LC3-II and p62 levels, indicating autophagosome accumulation and reduced autophagic flux. With transmission electron microscopy, we verified the increase in autophagosomes and also found irregularly shaped vesicular structures in Vps18 mutants. Subsequently, Vps18 mutant neutrophil progenitors underwent premature apoptosis. We described a novel patient with a heterozygous stop-gain mutation in VPS18 suffering from neutropenia and recurrent infections. To verify our findings in the human system, we used human induced pluripotent stem cells (iPSCs). Upon differentiation into neutrophils, loss of VPS18 resulted in an almost complete absence of iPSC-derived developing neutrophils. Heterozygous VPS18 mutant and patient mutation-harboring iPSCs were characterized by strongly reduced numbers of developing neutrophils. Zebrafish larvae with heterozygous mutations in vps18 were also characterized by significantly reduced neutrophil numbers. This study shows the pivotal impact of VPS18 for adequate vesicle dynamics during neutrophil development which might be relevant in the context of vesicle trafficking during granulopoiesis and congenital neutropenia.

physiology↗