bioRxiv Science⌕ Search
Home/Archive/bioRxiv

bioRxiv · 10.1101/2024.11.11.623018

Combined cellular and proteomics approach suggest differential processing of native and foreign vibrios in the sponge Halichondria panicea

Marulanda-Gomez, A. M.·Mueller, B.·Bayer, K.·Abukhalaf, M.·Tholey, A.·Fraune, S.·Pita, L.·Hentschel, U.
View original source ↗Export metadata ↓

Abstract

Phagocytosis is a conserved cellular mechanism for food uptake, defense and general animal-microbe interactions in metazoans. How the discrimination and subsequent processing of different microbes in marine invertebrates is facilitated remains largely unknown. Thereto, we combined a recently developed phagocytic assay with proteomics analysis to compare the phagocytic activity of the sponge Halichondria panicea upon encounter with a native (i.e., H. panicea isolate; Hal 281) and a foreign (i.e., Nematostella vectensis isolate; NJ 1) Vibrio. The sponge cell fraction was recovered after Vibrio exposure of 30 and 60 min and used for cellular (fluorescence-activated cell sorting and microscopy) and proteomics analyses. While the number of phagocytic active cells was similar between the isolates (p = 0.19), the distribution of vibrios over cell types differed (p = 0.02) with the tendency for a higher accumulation of foreign vibrios in small phagocytic cells than the native vibrios. Initially, both vibrios elicited a mitochondrial proteomic response (e.g., TBXASI, CASP7), whereas after 60 min differentially abundant proteins were dominated by lysosomal proteins (e.g., AGA, FUCA, SCARB2) in response to the native compared to immune-related proteins (e.g., AQP9, H2A, SAMHDI) in response to the foreign vibrio. H. panicea may discriminate native and foreign vibrios post-cellular internalization into choanocytes, followed by differential cellular and molecular processing: Native vibrios are readily transferred to archeocytes and elicit a digestive response, whereas foreign vibrios further accumulate before being transferred and provoke an immune response. These findings provide a mechanism for immune specificity in sponges. ImportanceMetazoans recognize and discriminate between native (i.e., associated and/or commonly encountered) and foreign microbes. In marine invertebrates, the mechanisms of microbial discrimination and immune specificity are not well understood. Phagocytosis is a conserved cellular process from amoeba to humans that facilitates the ingestion and digestion of microbial cells and likely plays a role in this discrimination. To elucidate the molecular and cellular basis of immune specificity in marine invertebrates, we examined the differential phagocytic processing of native and foreign Vibrio isolates in a marine sponge. Our findings revealed that both vibrios provoke a generalist mitochondrial protein-mediated response, while native vibrios trigger a digestive response, and foreign vibrios elicit an immune response after being internalized into sponge cells. This study uncovers a mechanism for immune specificity in an early-divergent metazoan and provides a valuable model for studying the evolution of immunity and its role in animal-microbe interactions.

Explore related subjects

cell biologyPreprint

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Marulanda-Gomez, A. M., Mueller, B., Bayer, K., Abukhalaf, M., Tholey, A., Fraune, S., Pita, L., Hentschel, U.. 2024-11-13. Combined cellular and proteomics approach suggest differential processing of native and foreign vibrios in the sponge Halichondria panicea. https://doi.org/10.1101/2024.11.11.623018

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Deep generative embeddings of gene expression and splicing reposition the interpretation of single-cell transcriptomic signatures

Single-cell transcriptomic analysis predominantly derives cell identity from gene expression analysis, while alternative splicing is processed separately despite its fundamental role for cell homeostasis. To overcome the limits of separate investigations, we developed a probabilistic deep learning framework, Crecerelle, enabling resolution of the contributions of gene expression and alternative splicing in each cell. Crecerelle learns cell embeddings from gene expressions and alternative splicing isoforms, to decipher their mutually dependent impact on the functional characterisation of cells in a data-driven manner, exemplified for the Tabula Muris dataset. This is enabled through a zero-and-N-inflated Dirichlet-Multinomial for a variational autoencoder that learns cell embeddings solely from splicing profiles, as well as a bi-modal variational autoencoder with a relevance-weighted mixture-of-experts variational posterior to consolidate the modality-specific contribution at single-cell level. Crecerelle reveals cell-type-specific isoform markers as well as subpopulations with unique isoforms and uncovers regulatory and disease-associated pathways not detected by gene expression analyses alone. This scalable and interpretable framework thus allows a more holistic study of transcriptomic regulation and will open a route to modality-relevance-weighted investigations across single-cell multiomics datasets and their influence on cellular homeostasis, tissue development and disease phenotypes.

cell biology↗
bioRxiv2026

MHC Molecules on B Cell Microvilli Are Spatially Associated with IL-15Rα

Interleukin-15 (IL-15) trans-presentation (TP) by B cells is an important mechanism of T-cell activation; however, the spatial organisation of interleukin-15 receptor (IL-15R) relative to major histocompatibility complex (MHC) molecules on B-cell microvilli remains poorly understood. As microvilli protrude from the B-cell surface and may serve as sites of initial B cell-T-cell contact, the distribution of IL-15R and MHC molecules within these structures may be important during the earliest stages of T-cell recognition and activation. Here, we investigated the spatial association and molecular proximity of IL-15R with MHC class I and class II molecules on B-cell microvilli before immunological synapse formation, using confocal microscopy, stimulated emission depletion (STED) microscopy, stochastic optical reconstruction microscopy (STORM), and fluorescence lifetime imaging microscopy-based Forster resonance energy transfer (FLIM-FRET). Both MHC class I and class II molecules showed significant spatial association with IL-15R; however, the extent of colocalisation decreased as spatial resolution increased. STED microscopy revealed significant colocalisation between IL-15R and MHC class I, whereas STORM did not detect this association. In contrast, IL-15R and MHC class II remained significantly colocalised at both resolutions. FLIM-FRET further demonstrated molecular proximity between IL-15R and both MHC class I and class II molecules, with higher FRET efficiency observed for MHC class II. Collectively, these findings indicate that IL-15R is spatially organised in proximity to both MHC class I and class II molecules on B-cell microvilli before immunological synapse formation. This arrangement at potential sites of initial B-cell-T-cell contact may facilitate the coordination of IL-15 trans-presentation and antigen presentation during the earliest stages of B-cell-T-cell interactions.

cell biology↗
bioRxiv2026

Pulsed-SILAC in single mouse embryos reveals early embryonic protein synthesis dynamics and phosphosite regulation

Early embryogenesis relies extensively on maternally deposited products until zygotic genome activation, yet the dynamics for the synthesis of new proteins in mammalian embryos remains poorly characterized. To address this, we applied pulsed stable isotope labelling by amino acids in cell culture (pSILAC) combined with narrow-window data-independent acquisition mass spectrometry to single mouse oocytes and embryos to resolve de novo protein synthesis during early embryogenesis. This revealed that the maternal proteome is not a static reservoir, with components of the subcortical maternal complex and amino acid transporters SLC7A1/2 being actively synthesized during the earliest developmental stages. Furthermore, phosphoproteomic analysis identified hundreds of previously unreported phosphosites and extensive regulation during the oocyte-to-embryo transition. Notably, phosphorylation of the PRC2-interacting KLP motif of EZHIP emerged as a potential regulatory mechanism, with modification of this region reducing EZHIP-PRC2 interaction and coinciding with H3K27me3 remodelling. Together, single embryo pSILAC revealed a maternal proteome that is continuously synthesized, recycled, and post-translationally regulated during early embryogenesis.

cell biology↗