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Perturbation of gut bacteria induces a coordinated cellular immune response in the purple sea urchin larva.

ABSTRACT: The purple sea urchin (Strongylocentrotus purpuratus) genome sequence contains a complex repertoire of genes encoding innate immune recognition proteins and homologs of important vertebrate immune regulatory factors. To characterize how this immune system is deployed within an experimentally tractable, intact animal, we investigate the immune capability of the larval stage. Sea urchin embryos and larvae are morphologically simple and transparent, providing an organism-wide model to view immune response at cellular resolution. Here we present evidence for immune function in five mesenchymal cell types based on morphology, behavior and gene expression. Two cell types are phagocytic; the others interact at sites of microbial detection or injury. We characterize immune-associated gene markers for three cell types, including a perforin-like molecule, a scavenger receptor, a complement-like thioester-containing protein and the echinoderm-specific immune response factor 185/333. We elicit larval immune responses by (1) bacterial injection into the blastocoel and (2) seawater exposure to the marine bacterium Vibrio diazotrophicus to perturb immune state in the gut. Exposure at the epithelium induces a strong response in which pigment cells (one type of immune cell) migrate from the ectoderm to interact with the gut epithelium. Bacteria that accumulate in the gut later invade the blastocoel, where they are cleared by phagocytic and granular immune cells. The complexity of this coordinated, dynamic inflammatory program within the simple larval morphology provides a system in which to characterize processes that direct both aspects of the echinoderm-specific immune response as well as those that are shared with other deuterostomes, including vertebrates.

SUBMITTER: Ch Ho E

PROVIDER: S-EPMC5073156 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature

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Perturbation of gut bacteria induces a coordinated cellular immune response in the purple sea urchin larva.

Ch Ho Eric E Buckley Katherine M KM Schrankel Catherine S CS Schuh Nicholas W NW Hibino Taku T Solek Cynthia M CM Bae Koeun K Wang Guizhi G Rast Jonathan P JP

Immunology and cell biology 20160519 9

The purple sea urchin (Strongylocentrotus purpuratus) genome sequence contains a complex repertoire of genes encoding innate immune recognition proteins and homologs of important vertebrate immune regulatory factors. To characterize how this immune system is deployed within an experimentally tractable, intact animal, we investigate the immune capability of the larval stage. Sea urchin embryos and larvae are morphologically simple and transparent, providing an organism-wide model to view immune r ...[more]

PMID: 27192936

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Project description:In this research we present a transcriptomics analysis of the physiological response of a marine calcifier, Strongylocentrotus purpuratus, to ocean acidification, a decline in ocean pH that results from the absorption of anthropogenic carbon dioxide (CO2). Larvae were raised from fertilization to prism stage in seawater with elevated CO2 conditions based upon IPCC emissions scenario B1 (540ppm CO2) and A1FI (1020ppm CO2). Adult S. purpuratus were collected using SCUBA from Goleta Pier (Goleta,CA, USA) and maintained in flowing seawater tables at 15-16°C. Spawning was induced by coelomic injection of 0.5M KCl following standard methods (Strathmann 1987). Eggs were collected from 3 females and separately fertilized by sperm from a single male (i.e. all larvae were full or half-siblings). Each of the three replicate cultures were divided in three (nine cultures total) and held in 10L flow-through containers filled with seawater aerated through a side arm with commercially manufactured air containing three different pCO2: 380ppm CO2 (present day atmospheric CO2 level, pH 8.01 ± 0.01), 540ppm CO2, an optimistic atmospheric CO2 concentration predicted by the Intergovernmental Panel on Climate Change for 2100 (B1 scenario, pH 7.96 ± 0.01) and 1020ppm CO2 (A1FI scenario, pH 7.88 ± 0.01). Larvae were cultured at 15°C until prism stage (40h post-fertilization). At the 40 hour time point, a sample of 60,000 larvae was removed from each of the 9 cultures in the 380, 540 and 1020 ppm treatments and stored in 1 mL Trizol at -80 ºC for later RNA analysis. The larval cDNA sample from each replicate 540ppm CO2 culture was competitively hybridized against the 380ppm CO2 control culture from the same replicate female. Similarly, the larval cDNA sample from each replicate 1020ppm CO2 culture was competitively hybridized against the 380ppm CO2 control culture from the same replicate female. Using dye swaps of technical replicates, treatment effects could be estimated independently of dye effects.

Dataset Information

Perturbation of gut bacteria induces a coordinated cellular immune response in the purple sea urchin larva.

Publications

Perturbation of gut bacteria induces a coordinated cellular immune response in the purple sea urchin larva.

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