A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions

Marulanda-Gómez, Ángela; Bayer, Kristina; Pita, Lucía; Hentschel, Ute

doi:10.3389/fmars.2023.1176145

Cited by 3 publications

(1 citation statement)

References 71 publications

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“…Long-term experiments in which sponges are evaluated after the filtering experiment to determine whether strains can be established in the host would also be helpful, although this may first require the development of axenic sponge models which has been attempted but remains unresolved (41,42). Alternatively, assays to study how sponges differentiate between food and symbionts, for example, may prove insightful (43).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of vectors for gene expression inPseudovibriobacteria and their application inAplysinamarine sponge studies

Dai,

Pita,

Eustáquio

2024

Preprint

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The filter feeding capacity of marine sponges contributes to biogeochemical cycling and they are also involved in habitat formation, properties that are critical to marine ecology. Sponge-associated microbes are crucial to the functional roles provided by sponges. α-Proteobacteria belonging to thePseudovibriogenus have been isolated from many different marine sponge genera and have been proposed to contribute to sponge health. We recently reported specialized metabolites we named pseudovibriamides fromPseudovibrio brasiliensisAb134. The pseudovibriamide encodingpppgene cluster is found in two thirds ofPseudovibriogenomes. Pseudovibriamides coordinate motility and biofilm formation, behaviors that are known to be important for host colonization. Although reverse genetics methods to delete genes via homologous recombination have been established, no self-replicative vectors have been reported forPseudovibrio. We show that plasmid vectors containing three different broad-host-range replicons, RSF1010, RK2, and pBBR1, can be used inP. brasiliensisfor fluorescent protein expression and consequent labeling. We then applied GFP and mCherry expressing strains to answer the question of whether pseudovibriamides affect the uptake ofP. brasiliensisbyAplysina aerophobasponges.P. brasiliensiscell counts decreased in the sponge aquaria at an equivalent rate for wild-type and pseudovibriamide-defective ΔpppAmutant strains, indicating that the sponge filters each strain indiscriminately under the conditions tested. Yet, the filtering capacity varied for each sponge individual tested, stressing the importance of performing experiments with wild-type and mutant bacterial strains in the same aquarium to allow for rigorous conclusions, which is now enabled with the methods established here.ImportanceMarine sponges are ecosystem engineers. They transform nutrients into a bioavailable form for other marine organisms. Microbes are critical to the functional roles provided by sponges because they expand the metabolic capabilities of the sponge host. Yet, most of our knowledge on sponge microbes comes from genomic studies, since cultivability and the ability to perform genetics with sponge bacterial isolates is limited. The genusPseudovibrioof α-Proteobacteria has consistently been isolated from marine sponges and it has been hypothesized to contribute to marine sponge health. Moreover,Pseudovibriobacteria are a source of antibiotics and other secondary metabolites with the potential to be developed into pharmaceuticals. Here we established vectors for the expression of fluorescent proteins inPseudovibriobacteria and demonstrated their utility inin vivostudies with marine sponges. The availability of genetic tools is important to enable us to explore the emerging ecological and biotechnological potential ofPseudovibriobacteria.

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Section: Discussionmentioning

confidence: 99%

Evaluation of vectors for gene expression inPseudovibriobacteria and their application inAplysinamarine sponge studies

Dai,

Pita,

Eustáquio

2024

Preprint

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The Buds of Oscarella lobularis (Porifera, Homoscleromorpha): A New Convenient Model for Sponge Cell and Evolutionary Developmental Biology

Rocher,

Vernale,

Fierro‐Constaín

et al. 2024

J Exp Zool Pt B

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The comparative study of the four non‐bilaterian phyla (Cnidaria, Placozoa, Ctenophora, and Porifera) provides insights into the origin of bilaterian traits. To complete our knowledge of the cell biology and development of these animals, additional non‐bilaterian models are needed. Given the developmental, histological, ecological, and genomic differences between the four sponge classes (Demospongiae, Calcarea, Homoscleromorpha, and Hexactinellida), we have been developing the Oscarella lobularis (Porifera, class Homoscleromorpha) model over the past 15 years. Here, we report a new step forward by inducing, producing, and maintaining in vitro thousands of clonal buds that now make possible various downstream applications. This study provides a full description of bud morphology, physiology, cells and tissues, from their formation to their development into juveniles, using adapted cell staining protocols. In addition, we show that buds have outstanding capabilities of regeneration after being injured and of re‐epithelization after complete cell dissociation. Altogether, Oscarella buds constitute a relevant all‐in‐one sponge model to access a large set of biological processes, including somatic morphogenesis, epithelial morphogenesis, cell fate, body axes formation, nutrition, contraction, ciliary beating, and respiration.

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Host control of the microbiome: Mechanisms, evolution, and disease

Wilde,

Slack,

Foster

2024

Science

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Many species, including humans, host communities of symbiotic microbes. There is a vast literature on the ways these microbiomes affect hosts, but here we argue for an increased focus on how hosts affect their microbiomes. Hosts exert control over their symbionts through diverse mechanisms, including immunity, barrier function, physiological homeostasis, and transit. These mechanisms enable hosts to shape the ecology and evolution of microbiomes and generate natural selection for microbial traits that benefit the host. Our microbiomes result from a perpetual tension between host control and symbiont evolution, and we can leverage the host’s evolved abilities to regulate the microbiota to prevent and treat disease. The study of host control will be central to our ability to both understand and manipulate microbiotas for better health.

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A novel in-vivo phagocytosis assay to gain cellular insights on sponge-microbe interactions

Cited by 3 publications

References 71 publications

Evaluation of vectors for gene expression inPseudovibriobacteria and their application inAplysinamarine sponge studies

Evaluation of vectors for gene expression inPseudovibriobacteria and their application inAplysinamarine sponge studies

The Buds of Oscarella lobularis (Porifera, Homoscleromorpha): A New Convenient Model for Sponge Cell and Evolutionary Developmental Biology

Host control of the microbiome: Mechanisms, evolution, and disease

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