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A novel F4-carbohydrate recognition domain (CRD)–linker–F3-CRD-type bi-CRD Branchiostoma belcheri tsingtauense galectin (BbtGal)-L together with its alternatively spliced mono-CRD isoform BbtGal-S from amphioxus intestine was encoded by a 9488-bp unique gene with eight exons and seven introns. The recombinant proteins of BbtGal were found to have β-galactoside-binding activity, indicating that BbtGal was a member of the galectin family. Phylogenetic analysis of this gene along with its splicing form and genome structure suggested that the BbtGal gene was the primitive form of the chordate galectin family. Real-time polymerase chain reaction analyses (PCR) indicated that BbtGal mRNA was expressed during all stages of embryonic development. In terms of tissue distribution, BbtGal-L mRNA was mainly expressed in the immunity-related organs, such as hepatic diverticulum, intestine, and gill, but BbtGal-S was ubiquitously expressed in all tissues. The expression of BbtGal-L mRNA was elevated after acute challenge with various microorganisms, but BbtGal-L only bound to specific bacteria. The immune function of BbtGal was consistent with its localization both outside and inside the cell. Our study on amphioxus galectin may help further understanding of the evolution of chordate galectin in terms of host–pathogen interaction in the immune system. [ABSTRACT FROM AUTHOR]

Both amphioxus and the sea urchin encode a complex innate immune gene repertoire in their genomes, but the composition and mechanisms of their innate immune systems, as well as the fundamental differences between two systems, remain largely unexplored. In this study, we dissect the mucosal immune complexity of amphioxus into different evolutionary-functional modes and regulatory patterns by integrating information from phylogenetic inferences, genome-wide digital expression profiles, time course expression dynamics, and functional analyses. With these rich data, we reconstruct several major immune subsystems in amphioxus and analyze their regulation during mucosal infection. These include the TNF/IL-1R network, TLR and NLR networks, complement system, apoptosis network, oxidative pathways, and other effector genes (e.g., peptidoglycan recognition proteins, Gram-negative binding proteins, and chitin-binding proteins). We show that beneath the superficial similarity to that of the sea urchin, the amphioxus innate system, despite preserving critical invertebrate components, is more similar to that of the vertebrates in terms of composition, expression regulation, and functional strategies. For example, major effectors in amphioxus gut mucous tissue are the well-developed complement and oxidative-burst systems, and the signaling network in amphioxus seems to emphasize signal transduction/modulation more than initiation. In conclusion, we suggest that the innate immune systems of amphioxus and the sea urchin are strategically different, possibly representing two successful cases among many expanded immune systems that arose at the age of the Cambrian explosion. We further suggest that the vertebrate innate immune system should be derived from one of these expanded systems, most likely from the same one that was shared by amphioxus. [ABSTRACT FROM AUTHOR]

It has been speculated that before vertebrates evolved somatic diversity-based adaptive immunity, the germline-encoded diversity of innate immunity may have been more developed. Amphioxus occupies the basal position of the chordate phylum and hence is an important reference to the evolution of vertebrate immunity. Here we report the first comprehensive genomic survey of the immune gene repertoire of the amphioxus Branchiostoma floridae. It has been reported that the purple sea urchin has a vastly expanded innate receptor repertoire not previously seen in other species, which includes 222 toll-like receptors (TLRs), 203 NOD/NALP-like receptors (NLRs), and 218 scavenger receptors (SRs). We discovered that the amphioxus genome contains comparable expansion with 71 TLR gene models, 118 NLR models, and 270 SR models. Amphioxus also expands other receptor-like families, including 1215 C-type lectin models, 240 LRR and IGcam-containing models, 1363 other LRR-containing models, 75 C1q-like models, 98 ficolin-like models, and hundreds of models containing complement-related domains. The expansion is not restricted to receptors but is likely to extend to intermediate signal transducers because there are 58 TIR adapter-like models, 36 TRAF models, 44 initiator caspase models, and 541 death-fold domain-containing models in the genome. Amphioxus also has a sophisticated TNF system and a complicated complement system not previously seen in other invertebrates. Besides the increase of gene number, domain combinations of immune proteins are also increased. Altogether, this survey suggests that the amphioxus, a species without vertebrate-type adaptive immunity, holds extraordinary innate complexity and diversity. [ABSTRACT FROM AUTHOR]