A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions
IntroductionSponges harbor diverse, specific, and stable microbial communities, but at the same time, they efficiently feed on microbes from the surrounding water column. This filter-feeding lifestyle poses the need to distinguish between three categories of bacteria: food to digest, symbionts to in...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-08-01
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| Series: | Frontiers in Marine Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmars.2023.1176145/full |
| _version_ | 1827871602941886464 |
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| author | Angela M. Marulanda-Gomez Kristina Bayer Lucia Pita Ute Hentschel Ute Hentschel |
| author_facet | Angela M. Marulanda-Gomez Kristina Bayer Lucia Pita Ute Hentschel Ute Hentschel |
| author_sort | Angela M. Marulanda-Gomez |
| collection | DOAJ |
| description | IntroductionSponges harbor diverse, specific, and stable microbial communities, but at the same time, they efficiently feed on microbes from the surrounding water column. This filter-feeding lifestyle poses the need to distinguish between three categories of bacteria: food to digest, symbionts to incorporate, and pathogens to eliminate. How sponges discriminate between these categories is still largely unknown. Phagocytosis is conceivable as the cellular mechanism taking part in such discrimination, but experimental evidence is missing. We developed a quantitative in-vivo phagocytosis assay using an emerging experimental model, the sponge Halichondria panicea.MethodsWe incubated whole sponge individuals with different particles, recovered the sponge (host) cells, and tracked the incorporation of these particles into the sponge cells. Fluorescence-activated cell sorting (FACS) and fluorescent microscopy were used to quantify and verify phagocytic activity, defined here as the population of sponge cells with incorporated particles. Sponges were incubated with a green microalgae to test if particle concentration in the seawater affects the percentage of phagocytic activity, and to determine the timing where the maximum of phagocytic cells are captured in a pulse-chase experiment. Lastly, we investigated the application of our phagocytic assay with other particle types (i.e., fluorescently-labelled bacteria and fluorescent beads).Results and discussionThe percentage of sponge cells that had incorporated algae, bacteria, and beads ranged between 5 to 24%. These phagocytic sponge cells exhibited different morphologies and sizes depending on the type of particle presented to the sponge. Particle incorporation into sponge cells was positively related to algal concentration in the seawater, suggesting that sponge cells adjust their phagocytic activity depending on the number of particles they encounter. Our results further revealed that sponge phagocytosis initiates within minutes after exposure to the particles. Fluorescent and TEM microscopy rectified algal internalization and potential digestion in sponge cells. To our knowledge, this is the first quantitative in-vivo phagocytosis assay established in sponges that could be used to further explore phagocytosis as a cellular mechanism for sponges to differentiate between different microorganisms. |
| first_indexed | 2024-03-12T16:12:30Z |
| format | Article |
| id | doaj.art-7159f79e3e064a97b4e464ec4fa401bb |
| institution | Directory Open Access Journal |
| issn | 2296-7745 |
| language | English |
| last_indexed | 2024-03-12T16:12:30Z |
| publishDate | 2023-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Marine Science |
| spelling | doaj.art-7159f79e3e064a97b4e464ec4fa401bb2023-08-09T12:24:19ZengFrontiers Media S.A.Frontiers in Marine Science2296-77452023-08-011010.3389/fmars.2023.11761451176145A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactionsAngela M. Marulanda-Gomez0Kristina Bayer1Lucia Pita2Ute Hentschel3Ute Hentschel4Research Unit Marine Symbioses, GEOMAR Helmholtz Centre for Ocean Research Kiel, Kiel, GermanyResearch Unit Marine Symbioses, GEOMAR Helmholtz Centre for Ocean Research Kiel, Kiel, GermanyInstitut de Ciències del Mar–Spanish National Research Council (CSIC), Marine Biology and Oceanography, Marine Biogeochemistry, Atmosphere and Climate, Barcelona, SpainResearch Unit Marine Symbioses, GEOMAR Helmholtz Centre for Ocean Research Kiel, Kiel, GermanyChristian-Albrechts-Universität Kiel, Kiel, GermanyIntroductionSponges harbor diverse, specific, and stable microbial communities, but at the same time, they efficiently feed on microbes from the surrounding water column. This filter-feeding lifestyle poses the need to distinguish between three categories of bacteria: food to digest, symbionts to incorporate, and pathogens to eliminate. How sponges discriminate between these categories is still largely unknown. Phagocytosis is conceivable as the cellular mechanism taking part in such discrimination, but experimental evidence is missing. We developed a quantitative in-vivo phagocytosis assay using an emerging experimental model, the sponge Halichondria panicea.MethodsWe incubated whole sponge individuals with different particles, recovered the sponge (host) cells, and tracked the incorporation of these particles into the sponge cells. Fluorescence-activated cell sorting (FACS) and fluorescent microscopy were used to quantify and verify phagocytic activity, defined here as the population of sponge cells with incorporated particles. Sponges were incubated with a green microalgae to test if particle concentration in the seawater affects the percentage of phagocytic activity, and to determine the timing where the maximum of phagocytic cells are captured in a pulse-chase experiment. Lastly, we investigated the application of our phagocytic assay with other particle types (i.e., fluorescently-labelled bacteria and fluorescent beads).Results and discussionThe percentage of sponge cells that had incorporated algae, bacteria, and beads ranged between 5 to 24%. These phagocytic sponge cells exhibited different morphologies and sizes depending on the type of particle presented to the sponge. Particle incorporation into sponge cells was positively related to algal concentration in the seawater, suggesting that sponge cells adjust their phagocytic activity depending on the number of particles they encounter. Our results further revealed that sponge phagocytosis initiates within minutes after exposure to the particles. Fluorescent and TEM microscopy rectified algal internalization and potential digestion in sponge cells. To our knowledge, this is the first quantitative in-vivo phagocytosis assay established in sponges that could be used to further explore phagocytosis as a cellular mechanism for sponges to differentiate between different microorganisms.https://www.frontiersin.org/articles/10.3389/fmars.2023.1176145/fullsponge-microbe symbiosisphagocytosisfluorescence-activated cell sorting (FACS)particle uptakesponge cells |
| spellingShingle | Angela M. Marulanda-Gomez Kristina Bayer Lucia Pita Ute Hentschel Ute Hentschel A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions Frontiers in Marine Science sponge-microbe symbiosis phagocytosis fluorescence-activated cell sorting (FACS) particle uptake sponge cells |
| title | A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions |
| title_full | A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions |
| title_fullStr | A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions |
| title_full_unstemmed | A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions |
| title_short | A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions |
| title_sort | novel in vivo phagocytosis assay to gain cellular insights on sponge microbe interactions |
| topic | sponge-microbe symbiosis phagocytosis fluorescence-activated cell sorting (FACS) particle uptake sponge cells |
| url | https://www.frontiersin.org/articles/10.3389/fmars.2023.1176145/full |
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