RESUMO
Sea anemones are known to produce a diverse array of toxins with different cysteine-rich peptide scaffolds in their venoms. The serine peptidase inhibitors, specifically Kunitz inhibitors, are an important toxin family that is believed to function as defensive peptides, as well as prevent proteolysis of other secreted anemone toxins. In this study, we isolated three serine peptidase inhibitors named Anthopleura cascaia peptide inhibitors I, II, and III (ACPI-I, ACPI-II, and ACPI-III) from the venom of the endemic Brazilian sea anemone A. cascaia. The venom was fractionated using RP-HPLC, and the inhibitory activity of these fractions against trypsin was determined and found to range from 59% to 93%. The spatial distribution of the anemone peptides throughout A. cascaia was observed using mass spectrometry imaging. The inhibitory peptides were found to be present in the tentacles, pedal disc, and mesenterial filaments. We suggest that the three inhibitors observed during this study belong to the venom Kunitz toxin family on the basis of their similarity to PI-actitoxin-aeq3a-like and the identification of amino acid residues that correspond to a serine peptidase binding site. Our findings expand our understanding of the diversity of toxins present in sea anemone venom and shed light on their potential role in protecting other venom components from proteolysis.
RESUMO
Background: Like all cnidarians, the subclass Ceriantharia (Cnidaria, Anthozoa) is known for producing cnidocysts, which mainly serve for prey immobilization, predator defense, and locomotion. Aim: The present study aimed to understand the variability of the cnidom, i.e., the inventory of all cnidocyst types, in the ceriantharians (tube anemones) Ceriantheomorphe brasiliensis (10 individuals) and Cerianthus sp. (seven individuals). Methods: In each individual, 30 intact cnidocysts of each identified type were measured in the following parts of tube anemones: marginal tentacles (four from each individual), labial tentacles (four from each individual), column, actinopharynx and metamesenteries. Each of these structures was divided into three levels (high, middle, and low) and the cnidom was analyzed. Statistical descriptive parameters (mean, standard deviation, minimum and maximum) of the sizes of all types of cnidocysts were calculated. The normality of the data for cnidocyst length was assessed using a Shapiro-Wilk test (α = 0.05). Based on the acceptance or rejection of the normality, either linear models or generalized linear models were used to evaluate variations in cnidocyst lengths. The normality of the cnidocyst length was tested by Shapiro-Wilk, and due to its rejection, generalized linear mixed models were applied to test the cnidocyst lengths variations. Results: The analysis of Ceriantheomorphe brasiliensis revealed 23 categories of cnidocysts, thereby expanding the understanding of its cnidome. The cnidoms of Ceriantheomorphe brasiliensis and Cerianthus sp. presented intraspecific variations, both qualitatively and in the lengths of cnidocysts. The cnidoms of the two species studied also showed qualitative intra-individual variations between different levels (high, middle, low) within each structure of the tube anemone (tentacles, actinopharynx, column and metamesenteries). Some cnidocyst types, such as atrichs from the column of C. brasiliensis, presented a length gradient along the column, from larger lengths at the "low" level to smaller lengths at the "high" level. Conclusions: The cnidom of a tube anemone could be better described if samples are taken at different levels of the structures, as observed in C. brasiliensis. In addition, we can conclude that the cnidocyst lengths of both C. brasiliensis and Cerianthus sp. present intraspecific variation, which is coincident with that observed in actiniarian sea anemones. Moreover, as main conclusion, this work also proved that individuals of tube anemone species could present qualitative intra-structure variations in both the cnidom and cnidocyst lengths. This characteristic appears as an exception in cnidom variations, and has so far not been recorded even in the most studied actiniarian sea anemones. Finally, the intra-structure cnidocyst variations could reveal different functions of the different levels of a particular body part of the organisms.
Assuntos
Antozoários , Anêmonas-do-Mar , Humanos , Animais , NematocistoRESUMO
Background: Ceriantharia is a subclass of the phylum Cnidaria, which comprises tube-dwelling marine invertebrates. This subclass is composed of three families, including Arachnactidae, with two known genera. Currently, the genus Arachnanthus has five valid species recorded from Australia, the Mediterranean Sea and both the Southern and Northern Pacific Ocean. However, at the moment, there is no record of organisms of this family from the South Atlantic Ocean. Besides that, the life cycle of any species of the genus Arachnanthus is known. The present study describes a new species of the genus Arachnanthus and its life cycle, based on specimens from Uruguay and South of Brazil. Methods: Larvae were collected by plankton net in Rio Grande-Brazil and the development and external morphology of these specimens were observed in the laboratory during two years, and subsequently described. Additionally, nine adult ceriantharians correspondent to the larvae from Rio Grande were collected in Uruguay and their external and internal anatomies, and cnidome were described. Results: Arachnanthus errans sp. nov. exhibited a free-swimming, short-lived cerinula larvae that spent short-time on the plankton. The larva developed into small and translucent polyps with a short actinopharynx, one pair of mesenteries attached to a siphonoglyph, and a medium first pair of metamesenteries. Further, the adult polyp displayed an unprecedented locomotion behavior in Ceriantharia that is first reported here, it can crawl under and in between the sediment.
Assuntos
Antozoários , Animais , Oceano Atlântico , Larva , Estágios do Ciclo de Vida , Oceano PacíficoRESUMO
Sea anemones are known to produce a diverse array of toxins with different cysteine-rich peptide scaffolds in their venoms. The serine peptidase inhibitors, specifically Kunitz inhibitors, are an important toxin family that is believed to function as defensive peptides, as well as prevent proteolysis of other secreted anemone toxins. In this study, we isolated three serine peptidase inhibitors named Anthopleura cascaia peptide inhibitors I, II, and III (ACPI-I, ACPI-II, and ACPI-III) from the venom of the endemic Brazilian sea anemone A. cascaia. The venom was fractionated using RP-HPLC, and the inhibitory activity of these fractions against trypsin was determined and found to range from 59% to 93%. The spatial distribution of the anemone peptides throughout A. cascaia was observed using mass spectrometry imaging. The inhibitory peptides were found to be present in the tentacles, pedal disc, and mesenterial filaments. We suggest that the three inhibitors observed during this study belong to the venom Kunitz toxin family on the basis of their similarity to PI-actitoxin-aeq3a-like and the identification of amino acid residues that correspond to a serine peptidase binding site. Our findings expand our understanding of the diversity of toxins present in sea anemone venom and shed light on their potential role in protecting other venom components from proteolysis.
RESUMO
Phylum Cnidaria comprises more than 10,000 species and around 10% of them are represented by sea anemones. These animals are underexplored sources of molecules, possessing structurally diverse toxins that can act over a diverse range of pharmacological targets, including enzymes. Sea anemones represent almost 96% of the manually annotated toxins from the phylum, but until now only 5% of its species have been studied about their toxin content. In the present work, the venoms of the sea anemones Anthopleura cascaia and Aulactinia veratra were studied and accessed through mass spectrometry analysis for searching serine peptidase inhibitors. The arsenal of toxins from both venoms was elucidated. Additionally, venom’s fractions were screened for inhibitory activity over trypsin, using time-course fluorescence-based kinetic assays or Mass spectrometry-based analysis. Beyond that, the spatial distribution of serine peptidase inhibitors in both sea anemones’ tissues were shown through Mass Spectrometry Imaging by MALDITOF. In the analysis of toxins composition, it was seen that A. cascaia venom presents at least three types of toxins: cytolysins, phospholipases and a toxin similar to natterin. For A. veratra, the classification based on blastp hit similarity and relying on domain architecture of the toxin’s sequences (translated transcripts) was performed. The thorough examination over toxins sequences led to the identification of 59 proteins and peptides belonging to 14 known toxin’s families of sea anemones and to the acknowledge of 20 peptides presenting 18 new cysteine scaffolds. The venom of this sea anemone mainly relies on neurotoxins from ShK-like, β-defensins, SCRiP, ICK, EGF-like types and on serine peptidase inhibitors from Kazal and Kunitz types. Furthermore, serine peptidase inhibitors from both venoms were isolated and present main distribution over tentacles, mesenterial filaments and pedal disc of these sea anemones, suggesting the preferential stock of these toxins. In conclusion, the methodological approaches applied in this study were able of identifying the presence of serine peptidase inhibitors on the venom and tissue of sea anemones through chromatographic techniques followed by enzymatic assays, and MALDI-Imaging.
O filo Cnidaria é composto por mais de 10.000 espécies e cerca de 10% destas são anêmonas-do-mar. Estes animais são considerados fontes subexploradas de moléculas, possuindo um diverso arsenal de toxinas que podem agir sobre diferentes alvos farmacológicos, incluindo enzimas. Toxinas de anêmonas-do-mar representam cerca de 96% das toxinas anotadas para o filo Cnidaria, embora apenas 5% de suas espécies tenham sido estudadas quanto à composição de toxinas até o momento. Neste trabalho elucidamos por espectrometria de massas a composição da peçonha das anêmonas Anthopleura cascaia e Aulactinia veratra, buscando a identificação de inibidores de serinopeptidases. O arsenal de toxinas para ambas anêmonas foi elucidado. Ainda, descrevemos as etapas de purificação envolvidas na busca de inibidores e a seleção destes candidatos por meio da inibição da atividade da tripsina, avaliada por duas técnicas distintas։ Cinética enzimática e Espectrometria de massas. Adicionalmente, descrevemos a localização de candidatos a inibidores no tecido das anêmonas através do Imageamento por espectrometria de massas. Na análise sobre a composição de toxinas destas anêmonas, vimos que a peçonha da A. cascaia apresentou a existência três tipos de toxinas incluindo citolisinas, fosfolipases e naterinas. Para a espécie A. veratra, a classificação de toxinas baseadas no blastp hit e na arquitetura de domínios das toxinas foi realizada. Esta análise revelou a presença de 59 proteínas e peptídeos pertencentes a 14 famílias de toxinas de anêmonasdo-mar; além do reconhecimento de 20 peptídeos apresentando 18 novos scaffolds de cisteínas. A peçonha desta anêmona é principalmente composto por neurotoxinas do tipo ShK-like, β-defensinas, SCRiP, ICK, EGF-like e inibidores de serinopeptidases. Os dados obtidos mostram que ambas anêmonas são ricas fontes de inibidores de serinopeptidases, especialmente tipo Kunitz e Kazal. Tais inibidores apresentam distribuição na região dos tentáculos, mesentério e disco pedal das anêmonas, o que pode indicar o estoque preferencial destas toxinas. E conclusão, o conjunto de abordagens metodológicas empregadas neste trabalho foi capaz de atender os objetivos propostos: identificar a presença de inibidores de serinopeptidases na peçonha e tecido de anêmonas, tanto por fracionamento cromatográfico seguido de ensaio enzimático, quanto por MALDI-Imaging.
RESUMO
The Puerto Velero Marina, located on the central Caribbean coast of Colombia, is an example of an artificial structure that serves as a substrate for a vulnerable community such as cnidarians (Hydrozoa and Anthozoa). This study provides the first assessment of corals and other cnidarians inhabiting artificial marine habitats in a marginal environment of the Caribbean of Colombia. The Puerto Velero Marina was built into a 7 km2 sand spit generated by sedimentation at the mouth of the Magdalena River. In this study were recorded 14 cnidarian species, among which were found 48 small colonies of scleractinian corals such as Porites, Siderastrea, Phyllangia, and Astrangia. This initial biodiversity assessment of the artificial structure provides a baseline that highlights the importance of further monitoring programs to identify non-native species that could reach this kind of hard structures.
Assuntos
Antozoários , Hidrozoários , Animais , Região do Caribe , Colômbia , EcossistemaRESUMO
The diversity of Ceriantharia is known from studies formally describing species from the late 18th Century onwards. However, no nomenclators including a list and discussion of all valid species have been produced since a list discussed by Carlgren in 1912. The present nomenclator presents a complete list of adult species of Ceriantharia of the World, including a discussion on each species. It includes the three families (Arachnactidae, Botrucnidiferidae, Cerianthidae) and the currently accepted 54 species based on their adult form. This study serves as a presentation of the "state-of-the-art" list of species of Ceriantharia, and includes a species identification key to support taxonomic identification. Additional in-depth species-by-species investigations for almost all cerianthid species is still needed, as the information available for most of these species is quite superficial.