Plasma proteome responses in zebrafish following λ-carrageenan-Induced inflammation are mediated by PMN leukocytes and correlate highly with their human counterparts

Abstract

Regulation of inflammation is a critical process for maintaining physiological homeostasis. The λ-carrageenan (λ-CGN) is a mucopolysaccharide extracted from the cell wall of red algae (Chondrus crispus) capable of inducing acute intestinal inflammation, which is translated into the production of acute phase reactants secreted into the blood circulation. However, the associated mechanisms in vertebrates are not well understood. Here, we investigated the crucial factors behind the inflammatory milieu of λ-CGN-mediated inflammation administered at 0, 1.75, and 3.5% (v/w) by i.p. injection into the peritoneal cavity of adult zebrafish (ZF) (Danio rerio). We found that polymorphonuclear leukocytes (neutrophils) and lymphocytes infiltrating the ZF peritoneal cavity had short-term persistence. Nevertheless, they generate a strong pattern of inflammation that affects systemically and is enough to produce edema in the cavity. Consistent with these findings, cell infiltration, which causes notable tissue changes, resulted in the overexpression of several acute inflammatory markers at the protein level. Using reversed-phase high-performance liquid chromatography followed by a hybrid linear ion-trap mass spectrometry shotgun proteomic approach, we identified 2938 plasma proteins among the animals injected with PBS and 3.5% λ-CGN. First, the bioinformatic analysis revealed the composition of the plasma proteome. Interestingly, 72 commonly expressed proteins were recorded among the treated and control groups, but, surprisingly, 2830 novel proteins were differentially expressed exclusively in the λ-CGN-induced group. Furthermore, from the commonly expressed proteins, compared to the control group 62 proteins got a significant (p < 0.05) upregulation in the λ-CGN-treated group, while the remaining ten proteins were downregulated. Next, we obtained the major protein-protein interaction networks between hub protein clusters in the blood plasma of the λ-CGN induced group. Moreover, to understand the molecular underpinnings of these effects based on the unveiled protein sets, we performed a bioinformatic structural similarity analysis and generated overlapping 3D reconstructions between ZF and humans during acute inflammation. Biological pathway analysis pointed to the activation and abundance of diverse classical immune and acute phase reactants, several catalytic enzymes, and varied proteins supporting the immune response. Together, this information can be used for testing and finding novel pharmacological targets to treat human intestinal inflammatory diseases.

Complement System Inhibitory Drugs in a Zebrafish (Danio rerio) Model: Computational Modeling

“…The zebrafish model has been considered for studying the complement system since some of the components involved in the activity and regulation of the pathways of this system have been identified and cloned, including the proteins C1q [ 51 ], C4 [ 52 ], factor B [ 53 ], C3 [ 54 ], MBL [ 55 ], factor H [ 56 ], properdin [ 57 ] and CD59 [ 58 ]. Other regulatory factors of the complement system have been observed in studies on the induction of inflammatory responses in zebrafish [ 59 ]. Furthermore, high-resolution genome mapping and comparisons with the human genome have shown that the human genome has a similarity of approximately 70% with orthologous genes of zebrafish, thus emphasizing the biological reliability of this model [ 60 ].…”

Cyberlindnera jadinii yeast as a functional protein source: Modulation of immunoregulatory pathways in the intestinal proteome of zebrafish (Danio rerio)

“…Proteins are the main components of the physiological and metabolic pathways, and the establishment of protein-protein interaction networks and proteome profiling allows the identification of new biomarkers, disease related pathways and pathological processes. Changes in blood plasma proteome have been investigated in Atlantic salmon ( Salmo salar ) [ 13 ] and zebrafish [ 18 ], but data on changes in the proteome of intestinal tissues following dietary challenge are scarce. Kiron, Kathiresan [ 19 ] investigated alterations in the intestine mucus proteome induced by a functional diet containing a feed additive (nucleotides, antioxidant, vitamins C and E, beta-glucan) in Atlantic Salmon.…”