Dataset Information

Using magnetic nanoparticles to explore symbiotic interactions

ABSTRACT: All plants and animals form symbiotic associations with microbes, yet many of the underlying mechanisms associated with these interactions remain uncharacterized. There are inherent challenges to studying the cellular and metabolic interactions between eukaryotes and their microbial symbionts, thus new methodologies that enable the discovery of symbiotic processes are continually needed. Here, we explored the use of magnetic nanoparticles (MNPs) as a tool to track aspects of the host innate immune response to symbionts under both ex vivo and in vivo conditions. The symbiotic association between the Hawaiian bobtail squid Euprymna scolopes and its bioluminescent partner Vibrio fischeri was used as a model to explore the potential of MNPs as non-toxic, manipulable agents to investigate aquatic symbiotic associations. Results suggest that host cells can be effectively labeled with MNPs under ex vivo conditions and that the particles can be visualized and tracked within the host animal in vivo using magnetic particle imaging. Proteomic and metabolomic analyses also revealed minimal changes to the host innate immune cells after uptake of MNPs in the presence and absence of V. fischeri. Together, these results suggest that MNPs have minimal biochemical impact on the host cells and can serve as an effective tool to explore aquatic symbiotic interactions.

INSTRUMENT(S):

ORGANISM(S): Euprymna Scolopes (hawaiian Bobtail Squid)

TISSUE(S): Hemocyte

SUBMITTER: Jen Liddle

LAB HEAD: Jeremy Balsbaugh

PROVIDER: PXD074352 | Pride | 2026-05-25

REPOSITORIES: Pride

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			Action	DRS
	20230211_jcl_NVijayan_Nyholm_156-HL_800ng_180min_Wat25cmBEH_1.raw	Raw
	20230211_jcl_NVijayan_Nyholm_156-O_800ng_180min_Wat25cmBEH_1.raw	Raw
	20230211_jcl_NVijayan_Nyholm_165-MA_800ng_180min_Wat25cmBEH_1.raw	Raw
	20230211_jcl_NVijayan_Nyholm_165-O_800ng_180min_Wat25cmBEH_1.raw	Raw
	20230211_jcl_NVijayan_Nyholm_166-HL_800ng_180min_Wat25cmBEH_1.raw	Raw

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Publications

Using magnetic nanoparticles to explore symbiotic interactions.

Guillen Matus Dulce G DG Koch Eric J EJ Vijayan Nidhi N Good Hayden J HJ Samples Robert M RM Rinaldi-Ramos Carlos M CM Arnold David P DP Nyholm Spencer V SV Balunas Marcy J MJ Foster Jamie S JS

Scientific reports 20260401 1

All plants and animals form symbiotic associations with microbes, yet many of the underlying mechanisms associated with these interactions remain uncharacterized. There are inherent challenges to studying the cellular and metabolic interactions between eukaryotes and their microbial symbionts, thus new methodologies that enable the discovery of symbiotic processes are continually needed. Here, we explored the use of magnetic nanoparticles (MNPs) as a tool to track aspects of the host innate immu ...[more]

PMID: 41922716

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Project description:The squid, Euprymna scolopes, and the luminescent bacterium, Vibrio fischeri, form a highly specific beneficial light organ symbiosis. Not only does the host have to select V. fischeri from the environment, but it must also prevent subsequent colonization by non-symbiotic microorganisms. Host macrophage-like hemocytes are believed to play a role in mediating the symbiosis with V. fischeri. Previous studies have shown that the colonization state of the light organ influences the host’s hemocyte response to the symbiont. To further understand the molecular mechanisms behind this process, two quantitative mass spectrometry-based proteomic techniques, isobaric tags for relative and absolute quantification (iTRAQ) and label-free spectral counting, were used to compare and quantify the adult hemocyte proteomes from colonized (sym) and uncolonized (antibiotic-treated/cured) squid. Overall, iTRAQ allowed for the quantification of 1,024 proteins with 2 or more peptides. Thirty-seven unique proteins were determined to be significantly different between sym and cured hemocytes (p-value < 0.05), with 20 more abundant proteins and 17 less abundant in sym hemocytes. The label-free approach resulted in 1,241 proteins that were identified in all replicates. Of 185 unique proteins present at significantly different amounts in sym hemocytes (as determined by spectral counting), 92 were more abundant and 93 were less abundant. Comparisons between iTRAQ and spectral counting revealed that 30 of the 37 proteins quantified by iTRAQ exhibited similar trends identified by the label-free method. Both proteomic techniques mutually identified 16 proteins that were significantly different between the two groups of hemocytes (p-value <0.05). The presence of V. fischeri in the host light organ influenced the abundance of proteins associated with the cytoskeleton, adhesion, lysosomes, proteolysis, and the innate immune response. These data provide evidence that colonization by V. fischeri alters the hemocyte proteome and reveals proteins that may be important for maintaining host-symbiont specificity.

Project description:Beneficial microbial symbionts are often horizontally acquired by their animal hosts from environmental sources, requiring the symbionts to complete a lifestyle transition from free-living in the environment to association with host tissues. In the model symbiosis between the Hawaiian bobtail squid and its microbial symbiont Vibrio fischeri, one mechanism used to make this transition during host colonization is the formation of biofilm-like aggregates on host mucosa. Extensive work has previously been conducted to isolate the critical factors controlling V. fischeri biofilm formation, yet much remains unknown regarding the full breadth of the biofilm-associated regulon. Here, we probed in vitro models of biofilm formation using transcriptomics, to identify novel regulatory pathways active within biofilms of the V. fischeri type strain ES114. Through comparing the gene-sets which became differentially regulated in multiple biofilm models, we discovered a shared set of 232 genes which demonstrated similar patterns in expression relative to uninduced controls. These genes contained representatives of multiple exopolysaccharide loci, genes involved in flagellar motility, and a diverse collection of other genes. Follow-up analysis suggested that these transcriptomic changes reflected true phenotypic effects, including changes in motility and cyclic-di-GMP production in biofilm-induced backgrounds. Beyond characterizing the shared biofilm response, we additionally profiled the regulatory activity of the sensor kinase RscS. This sensor kinase has previously been characterized to function as a phospho-donor within an established biofilm-inducing phospho-relay, yet our data suggests that RscS moonlights in at least one other phospho-relay that integrates downstream signaling from a homolog of the Vibrio cholerae response regulator VpsR, without a need for its established signaling partners. Overall, this study adds to our understanding of the genes involved in V. fischeri biofilm regulation, while revealing new regulatory pathways branching from previously characterized signaling networks.