L panorama of the transcripts responsible for the physiological events that
L panorama of the transcripts responsible for the physiological events that take place in the venom gland, since those data were generated from a non-normalized cDNA library. Although there are many sequences from other fish in databases ?such as Takifugu rubripes, D. rerio, S. salar, Oryzias latipes and Gasterosteus aculeatus ?this is the first description of a fish venom gland transcriptome, which will help to support comparative studies for other fish venom glands. The primary structure of natterins was PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28607003 obtained by a transcriptomic approach using a representative cDNA library constructed from T. nattereri venom glands. First, several ESTs were obtained and processed by bioinformatics tools and revealed a major group (18 ) of related sequences unknown in gene or protein sequence databases. This group that included sequences showing the N-terminus of isolated natterins was called the natterin family. Analysis of this family allowed us to identify five related sequences, which we named 1? natterin and P-natterin. Natterin 1 and 2 sequences include the Nterminus of the isolated natterins (Figure 4). Furthermore,Previous work on the biochemical characterization and molecular identification of venom PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26866270 toxins, and evaluation of the inflammatory response in tissue injured by T. nattereri venom led to the construction of a methodological tool. It was useful for a better understanding of the RR6 supplier importance of the innate cellular response in the control of toxins and consequently in restoration of the integrity of damaged tissue. The injection of venom into footpad of mice increases the mRNA for IL-1, IL-6 and TNF- genes as well as the release of these soluble cytokines. Moreover, absence of leukocytes infiltration in the intraplantar region of footpad of mice after venom application was also reported [15]. Moreover, Pareja-Santos et al. [16] showed that T. nattereri venom alters the extracellular matrix structure of mouse footpad tissue by the activation of matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9), and it decreases collagen fiber production during the healing phase. It was also shown that the venom affects the cytoskeleton organization and pseudopodia formation of epithelial cells in in vitro system. Recently, Komegae et al. [17] demonstrated the specific ability of nattectin to bind types I and V collagens and natterins to bind and cleave type I collagen as well as type IV collagen, disrupting cell attachment and HeLa cells survival. Natterins have cytotoxic effect on both adherent cells or on those in suspension, showing direct induction of necrosis that is followed by cell detachment. Nattectin improves integrin-mediated HeLa cell adhesion and resistance to apoptosis by its binding to RGDdependent integrins, especially the 1 subunit. Based on our studies, we reported that extracellular matrix (ECM) components as well as the integrin 1 subunit areLopes-Ferreira et al. Journal of Venomous Animals and Toxins including Tropical Diseases 2014, 20:35 http://www.jvat.org/content/20/1/Page 7 ofFigure 4 Primary structure of toxins was obtained by a transcriptomic approach using a representative cDNA library constructed from T. nattereri venom glands. This approach also allowed us to prove the existence of natterins (1? and P), a novel family of kininogenases, and a toxic C-type lectin nattectin.targets for natterins and nattectin. The ECM degradation or remodeling activities exerted by these toxins affect cell-cell and cell-ECM adhesion and sur.