Journal: Biomolecules
Article Title: The Zebrafish Perivitelline Fluid Provides Maternally-Inherited Defensive Immunity
doi: 10.3390/biom10091274
Figure Lengend Snippet: ( A ) The Venn diagram shows the number of exclusive and shared proteins present in the perivitelline fluid (PVF) at different stages: early blastula (yellow), at 1 dpf (purple) and 2 dpf (green). The 85 sequences present at all developmental stages are considered as constitutive of PVF composition. ( B ) Enriched protein classes (according to their Gene Ontology) detected in the PVF. The overrepresentation analysis reveals a marked overrepresentation of protein classes related to immune defense at all stages (Fisher’s test; p < 0.05). ( C ) Identity of proteins (or gene ID) present in the PVF during embryonic development and their presence (purple) or absence (yellow) at different stages. The presence of a protein at 0 dpf suggests a maternal origin, since samples were taken before the MBT; the absence of a protein at this stage suggests a zygotic origin, while the presence before and after MBT, suggests a mixed maternal-zygotic origin. The gene list associated with each protein found in the PVF was organized by their protein superfamilies, families, and/or molecular functions. For example, the A2M superfamily corresponds to the “serine protease inhibitors” in B. ( D ) Zebrafish PVF can agglutinate live bacterial cells. The images show in vitro agglutination assays using fluorescent E. coli and S. typhimurium , which cluster in the presence of PVF. ( E ) PVF protects the zebrafish embryo from bacterial infection. Zebrafish embryos challenged at (i) 1 dpf and (ii) 2 dpf. The result reveals an augmented survival rate in the presence of PVF (Kruskal–Wallis test: **** p < 0.0001; *** p < 0.001; * p < 0.05). ( F ) The three main elements of early embryonic defense against pathogens in fish. The PVF constitutes the central element in a tripartite system of embryonic defenses against environmental pathogens. If an organism, chemical compound, or even a physical element from the exterior succeeds in crossing the chorion, it still must pass through the PVF to reach the embryo.
Article Snippet: Escherichia coli (DH5α) and Salmonella typhimurium (ΔaroA) transformed with the pDiGc vector (#59322 Addgene) to obtain GFP (green fluorescent protein) constitutive expression, were cultured on LB (Luria-Bertani) medium with ampicillin (100 μg/mL) at 37 °C.
Techniques: In Vitro, Agglutination, Infection
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