Identification of Transcripts Expressed Preferentially in Hemocytes of Ciona intestinalis that can be Used as Molecular Markers

Wakoh, Takeshi; Ikeda, Makoto; Uchino, Ryuji; Azumi, Kaoru; Nonaka, Masaru; Kohara, Yuji; Metoki, Hitoe; Satou, Yutaka; Satoh, Nori; Satake, Masanobu

doi:10.1093/dnares/11.5.345

Cited by 14 publications

(9 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…BM Purple (Roche) was used for the signal detection. ISH was carried out as described previously (44). IHC sections were incubated with anti-VCBP-C (2.0 μg/mL) overnight at 4°C and detected with an Vectastain Elite ABC Kit (Vector Laboratories) and Fast DAB (Sigma).…”

Section: Methodsmentioning

confidence: 99%

A role for variable region-containing chitin-binding proteins (VCBPs) in host gut–bacteria interactions

Dishaw

Giacomelli²,

Melillo³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

114

View full text Add to dashboard Cite

A number of different classes of molecules function as structural matrices for effecting innate and adaptive immunity. The most extensively characterized mediators of adaptive immunity are the immunoglobulins and T-cell antigen receptors found in jawed vertebrates. In both classes of molecules, unique receptor specificity is effected through somatic variation in the variable (V) structural domain. V region-containing chitin-binding proteins (VCBPs) consist of two tandem Ig V domains as well as a chitin-binding domain. VCBPs are encoded at four loci (i.e., VCBPA-VCBPD) in Ciona, a urochordate, and are expressed by distinct epithelial cells of the stomach and intestine, as well as by granular amoebocytes present in the lamina propria of the gut and in circulating blood. VCBPs are secreted into the gut lumen, and direct binding to bacterial surfaces can be detected by immunogold analysis. Affinity-purified native and recombinant VCBP-C, as well as a construct consisting only of the tandem V domains, enhance bacterial phagocytosis by granular amoebocytes in vitro. Various aspects of VCBP expression and function suggest an early origin for the key elements that are central to the dialogue between the immune system of the host and gut microflora.bacteria opsonization | bacteria phagocytosis | immunoglobulin variable domains D ifferent molecular and cellular mechanisms that effect "innate" or "adaptive" immune responses shape immunity to pathogens such as viruses, bacteria, and parasites in all metazoans. The innate immune system includes germline-encoded receptor molecules that recognize widely divergent molecular structures. In contrast, the gene loci that encode the receptor molecules of the adaptive immune system undergo unique rearrangements in individual somatic cells that expand clonally and account for nearlimitless functional variation of receptors. The adaptive immune response is limited to jawed vertebrate species, whereas innate immunity is characteristic of all metazoan phyla.Alternative mechanisms of innate and adaptive immunity have been described in jawless vertebrates, protochordates, and other invertebrates (1, 2). Given the absence of V domain-mediated immunity in jawless vertebrates, the protochordate lineages are particular significant for understanding the origins of V region diversity as a basic form of immune recognition. In the protochordate Branchiostoma floridae (amphioxus), variable region-containing chitin-binding proteins (VCBPs) were described that consist of two variable (V) Ig domains and a single chitin-binding domain (3). VCBPs are encoded by diverse, nonrecombining, haplotypically variable alleles (4-6). Detailed structural studies of VCBPs reveal that the hyperpolymorphic positions are localized on the β-sheet surfaces of the folded V domains (7) and not on the connecting loops, which are the sites of the highest variability in the V domains of Ig and T-cell receptor (TCR). Other innate immune functions (e.g., viral receptor and superantigen-binding sites) are associated with V regi...

show abstract

Section: Methodsmentioning

confidence: 99%

A role for variable region-containing chitin-binding proteins (VCBPs) in host gut–bacteria interactions

Dishaw

Giacomelli²,

Melillo³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

114

View full text Add to dashboard Cite

show abstract

“…Oligonucleotide PCR primers pairs were designed based on FGENESH output sequences. Juvenile C. intestinalis cDNA was prepared as described (47). cDNA sequences were amplified from juvenile cDNA by PCR with Advantage 2 Polymerase (Clontech, Mountain View, CA) using reaction conditions recommended by the manufacturer.…”

Section: Methodsmentioning

confidence: 99%

“…Template cDNA was prepared as described (47). Exon-spanning primer pairs were designed for Ci-Gla1, Ci-Gla2, Ci-Gla3, Ci-Gla4, Ci-GGC, and Ci-VKOR based on the DNA sequences of cloned cDNAs.…”

Section: Analysis Of Gene Expression By Qpcrmentioning

confidence: 99%

Vitamin K-dependent proteins in Ciona intestinalis , a basal chordate lacking a blood coagulation cascade

Kulman

Harris

Nakazawa

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

We have isolated and sequenced several cDNAs derived from the sea squirt Ciona intestinalis that encode vitamin K-dependent proteins. Four of these encode ␥-carboxyglutamic acid (Gla) domain-containing proteins, which we have named Ci-Gla1 through Ci-Gla4. Two additional cDNAs encode the apparent orthologs of ␥-glutamyl carboxylase and vitamin K epoxide reductase. Ci-Gla1 undergoes ␥-glutamyl carboxylation when expressed in CHO cells and is homologous to Gla-RTK, a putative receptor tyrosine kinase previously identified in a related ascidian. The remaining three Gla domain proteins are similar to proteins that participate in fundamental developmental processes, complement regulation, and blood coagulation. These proteins are generally expressed at low levels throughout development and exhibit either relatively constant expression (Ci-Gla1, ␥-glutamyl carboxylase, and vitamin K epoxide reductase) or spatiotemporal regulation (Ci-Gla2, -3, and -4). These results demonstrate the evolutionary emergence of the vitamin K-dependent Gla domain before the divergence of vertebrates and urochordates and suggest novel functions for Gla domain proteins distinct from their roles in vertebrate hemostasis. In addition, these findings highlight the usefulness of C. intestinalis as a model organism for investigating vitamin K-dependent physiological phenomena, which may be conserved among the chordate subphyla.ascidian ͉ ␥-carboxyglutamic acid ͉ urochordate ͉ warfarin

show abstract

“…Although C6-like molecules have been described for amphioxus (Suzuki et al, 2000) and Ciona (Wakoh et al, 2004), their role as complement proteins remains unconfirmed. Molecular analysis of the C6-like gene described for amphioxus reveals a 5' C6 modular structure with a 3' end that is missing key modules characteristic of C6.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, in Ciona the C6-like gene is expressed as a cell surface receptor and whether it has a complement function (Wakoh et al, 2004;Azumi et al, 2003) in either organism is unknown. This amphioxus C6 gene has not been shown to be expressed or functional and may be C6-like only in sequence.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of GcC8 alpha, the functional homologue of human C8 alpha in the shark, Ginglymostoma cirratum

Aybar¹

View full text Add to dashboard Cite

show abstract

Identification of Transcripts Expressed Preferentially in Hemocytes of Ciona intestinalis that can be Used as Molecular Markers

Cited by 14 publications

References 24 publications

A role for variable region-containing chitin-binding proteins (VCBPs) in host gut–bacteria interactions

A role for variable region-containing chitin-binding proteins (VCBPs) in host gut–bacteria interactions

Vitamin K-dependent proteins in Ciona intestinalis , a basal chordate lacking a blood coagulation cascade

Molecular characterization of GcC8 alpha, the functional homologue of human C8 alpha in the shark, Ginglymostoma cirratum

Contact Info

Product

Resources

About