Sodium current was blocked in stellate-ganglion cells of both S. officinalis and O. rubescens by subfraction 8 peptide and raw venom (duct and milked), respectively, at dilutions similar to those found to be effective on squid neurons. Gaps have been introduced manually to force alignment to the conserved cysteines (in bold) and to maximize amino acid identity in the intra-cysteine loops (shaded areas). Sepia officinalis; skin coloration; defense; predator--prey; visual perception; Animal coloration plays a key role in many facets of natural and sexual selection ().Camouflage is a widespread phenomenon throughout nature and an important antipredator tactic (2, 3).Camouflaged animals use diverse body patterns to make detection or recognition more difficult (). Search facet . Although the combination of leader sequence and Cys-framework has been used to define at least 15 superfamilies (Olivera and Cruz, 2001; Terlau and Olivera, 2004), the leader sequence is of primary importance (Halai and Craik, 2009). (B) Effect of the subfraction A on INa in a squid neuron. The number of fragments observed was much greater after reduction–alkylation as fragmentation is hindered by intact disulfide bonds. Sepia officinalis Linn's Active constituents: No information regarding Sepia officinalis Linn's antidot is currently available. These toxins block both gastropod and vertebrate Na+ channels and also affect Ca2+ channels at high concentration (Daly et al., 2004). This consists of an initial scan of the full range from 200–2000 mass-to-charge ratio (m/z), a subsequent data-dependent zoomed scan of the 20 m/z window surrounding a precursor ion to decipher charge states, and a final fragmentation scan of the same precursor. This alignment is not shared by cal12a. S. officinalis eat small molluscs (snails, clams, etc), crabs, shrimps and other cuttlefish (!). Predicted primary structures of Cal12.1.1a and Cal12.2a in comparison with selected O-type toxins from other Conus species. The alignment has been centered on the vicinal cysteines and a single gap introduced in the MrVIB sequence. Thank you for your interest in spreading the word on Journal of Experimental Biology. At least 90% of the K+ channels under the currents analyzed are of one type that is blocked by TsTX-Kα, a peptide toxin isolated from the scorpion, Tityus serrulatus (Mathes et al., 1997; Ellis et al., 2001), and no consistent effect on these squid K+ channels was thus observed. 10C) is disulfide-bonded to C4, and the intervening region is thought to be important in conferring target specificity (Daly et al., 2004). Tentacular club with 5 or 6 suckers in each transverse row, the median ones moderately enlarged. Total venom for physiological experiments (Monterey specimens) was extruded from an excised venom duct onto a small piece of plastic film and suspended in 0.5 ml of external recording solution (see below), manually homogenized, heated to 90°C for 5 min and centrifuged at 12,000 g for 15 min. 1). of fractions found in 37 individual snails. Biol. Sciencia Marina 54, 375-388. Its habitat ranges from subtidal waters to depths of 200 meters. With supporting detail drawings A-F. A second-stage chromatographic separation of the active peptides eluting between 43–46 min with a shallow gradient allowed clean separation of two major peaks (peaks A and B in inset to Fig. An additional source of variation in peptide composition should be considered for C. californicus. 10A). The cephalopod is presented with the two long tentacles extended, therefore a dead specimen. The ability of the cuttlefish Sepia officinalis to change its visual appearance allows us to compare the animal's choice of patterns during movement to the predictions of models of motion camouflage. Anim Cogn 8 2005: 27–30. See on tasuta kasutada ja iga artikli või dokumendi saab alla laadida. Venom components of C. californicus and their effects on voltage-gated Na+ currents (INa) at 0 mV in squid neurons using the whole-cell patch-clamp technique. The reaction was allowed to proceed in the dark for 1 h at room temperature, after which the reaction was quenched by adding 10 μl of 200 mmol l–1 dithiothreitol. The common procedure was to add 200 μl of this solution to the venom, sonicate the sample for 10 min, vortex, and then centrifuge the insoluble material into a pellet at 12,000 g for 5 min. Abstract. Potential evolutionary significance of introns, Diversity and evolution of conotoxins based on gene expression profiling of, Identified ion channels in the squid nervous system, A mitogenic peptide amide encoded within the E peptide domain of the insulin-like growth factor IB prohormone, Megafauna of the upper miocene castaic formation, Los Angeles county, California, Analysis of bromotryptophan and hydroxyproline modifications by high-resolution, high-accuracy precursor ion scanning utilizing fragment ions with mass-deficient mass tags. Diversity in toxin structure and specificity is reflected in the feeding behavior of different Conus species. This separation targeted the late-eluting species, including those constituting the doublet, as described above. Identification of the residue, L-6-bromotryptophan, in peptides from, Post-translationally modified neuropeptides from, Novel gamma-carboxyglutamic acid-containing peptides from the venom of, Differentiation of venoms of predatory marine gastropods: divergence of orthologous toxin genes of closely related, Species-level phylogeography and evolutionary history of the hyperdiverse marine gastropod genus, Ecological release and venom evolution of a predatory marine snail at Easter Island, Molecular genetics of ecological diversification: duplication and rapid evolution of toxin genes of the venomous gastropod, Evolutionary diversification of multigene families: allelic selection of toxins in predatory cone snails, Gene expression and feeding ecology: evolution of piscivory in the venomous gastropod genus, Origins of diverse feeding ecologies within, Geographic variation in venom allelic composition and diets of the widespread predatory marine gastropod, Conotoxin modulation of voltage-gated sodium channels, Cholinergic receptor in Aplysia neurons: activation by a venom component from the marine snail Conus californicus, Venom of marine snail Conus californicus: biochemical studies of a cholinomimetic component, Interaction of a toxin from the scorpion Tityus serrulatus with a cloned K+ channel from squid (sqKv1A), Venomous cone snails: molecular phylogeny and the generation of toxin diversity, Potassium channels: from scorpion venoms to high-resolution structure, Control of the spatial distribution of sodium channels in giant fiber lobe neurons of the squid, Fast and slow activation kinetics of voltage-gated sodium channels in molluscan neurons, Evolution of voltage-gated Na(+) channels, KNOTTIN: the knottin or inhibitor cystine knot scaffold in 2007, Agitoxin footprinting the shaker potassium channel pore, Isolation and characterization of three novel Gla-containing, Conotoxins of the O-superfamily affecting voltage-gated sodium channels, Sequencing and mass profiling highly modified conotoxins using global reduction/alkylation followed by mass spectrometry, Intraspecific variation of venom injected by fish-hunting, Screening for post-translational modifications in conotoxins using liquid chromatography/mass spectrometry: an important component of conotoxin discovery, Bromocontryphan: post-translational bromination of tryptophan, Direct cDNA cloning of novel conopeptide precursors of the O-superfamily, muO conotoxins inhibit NaV channels by interfering with their voltage sensors in domain-2, Proteomic analysis of post-translational modifications, Fast inactivation of delayed rectifier K conductance in squid giant axon and its cell bodies, Spatial localization of calcium channels in giant fiber lobe neurons of the squid (Loligo opalescens), Cone venom – from accidental stings to deliberate injection, A new family of conotoxins that blocks voltage-gated sodium channels, The charybdotoxin family of K+ channel-blocking peptides, A novel conotoxin framework with a helix-loop-helix (Cs alpha/alpha) fold, Speciation of cone snails and interspecific hyperdivergence of their venom peptides. Presented by Médi-T. SEPIA OFFICINALIS. Sepia officinalis Linnaeus, 1758. kingdom Animalia > phylum Mollusca > class Cephalopoda > subclass Coleoidea > superorder Decapodiformes > order Sepiida > family Sepiidae > genus Sepia > species Sepia officinalis. Cysteines defining framework 12 and absolutely conserved amino acids are indicated by the darker gray shading. Given the strong structural constraints that are likely to be imparted by the absolutely and/or highly conserved regions of Cal12.1 toxins (i.e. (N=3). Sepia / Sep. 4. If diversity in the Cal12.1 family does involve recombination in vivo, this particular family must be unusually susceptible to this mechanism, because Cal12.2 sequences show no sign of the modular type of variation evident in the Cal12.1 family. 7. Sepia officinalis. Seven fractions were collected manually and used for bioassays as described in ‘Electrophysiology’ below. Although the approximate time of elution had previously been established for the active fraction of interest, differences in LC parameters employed between systems required another identification of the active peptides. 8 provides an alignment of the toxin-coding regions for 15 Cal12.1 variants that were found in more than one snail, or in separate amplifications from a single snail. As depicted in Fig. Man verwandte ihn als erwärmendes, menstruationsförderndes, fieber und harntreibendes Mittel. Conotoxin precursors contain a hydrophobic leader (signal) sequence and pro-peptide region, both of which are removed by proteolysis during biosynthesis, and the primary structure of the signal sequence is generally highly conserved between toxins having identical Cys-frameworks. Information provided on this Web site is neither intended nor implied to be a substitute
We have identified the toxin-coding region of Cal12.1.1a in 17 (of 37) individual snails analyzed by RT-PCR (see below) and in 15 clones derived from PCR amplifications of pooled RNA. However, significant identity does exist in the N-terminal leader sequence between Cal12.1 and Cal12.2 members and a number of framework 6/7 (C-C-CC-C-C) conotoxins (Fig. and W.F.G.). Such a loop could potentially present this segment of charged residues as a reaction surface that is critical to target specificity, similar to the situation thought to exist in MrVIB. Er diente als Milzpflanze, gegen die Pest, bei Rückenschmerzen, Husten und Augenleiden. It is prepared from the dried liquid contained in the ink bag of cuttlefish. and Ushira – Vetiver – Vetiveria zizanioides The bone may be of oblong, elliptical or oval shaped flat skeleton, hard and brittle, highly pulverisable. Lengths of the inter-Cys segments in the Cal12 sequences fit within the range of the O-superfamily, with the exception of the C1-C2 segment. Variation in the Cal12.1 family is clearly different from that in Cal12.2, where it may represent allelic variation (Duda and Palumbi, 2000; Duda and Lee, 2009; Duda et al., 2009) or the diversity among paralogous members of a gene family that have been subject to gene duplication and rapid evolution (Duda and Palumbi, 1999; Conticello et al., 2001; Duda, 2008; Puillandre et al., 2010). In most cases RNA was extracted from the venom duct plus its venom content, because this procedure substantially increased the yield of RNA (our unpublished data). A second-stage purification was carried out using a C18 narrow-bore column (2.0 mm) with a 1% gradient of solvent B and a flow rate of 250 μl min–1 and monitored at 210 nm. Fraction B (right) contains purified cal12b at 5194 Da. At least 15 homologues of these toxins were found by molecular cloning, and constitute the Cal12.1 family of cDNAs. cannot be held responsible for your actions nor any conditions resulting thereof. FAO, FI and 18 Sep 1978 Santa Cruz de Tenerife (Canary Islands) Meeting of the Ad Hoc Working Group on the Assessment of Cephalopod Stocks (Sepia officinalis )(Linn é, 1758.) This leads us to propose a nomenclature for members of the Cal12.1.1–3 subtypes by denoting individual members of a subtype by a letter suffix. ), Kandeksu (Saccharum spontaneum Linn. Enter multiple addresses on separate lines or separate them with commas. It preys primarily on molluscs and worms, with gastropods and bivalves being the major prey types in one reported study of gut contents (Kohn, 1966). Helen Skaer, Mike O'Donnell and Julian Dow remember Simon in their affectionate Obituary. Täältä saat lunastaa bonuksen ja ilmaiskierroksia sekä lukea lisää kasinosta. Unfortunately the only gut-content analysis reported for C. californicus was based on La Jolla specimens. Voltage-clamp experiments were carried out with neurons from other cephalopods as well as gastropods (data not illustrated). Leader and propeptide sequences are identical for Cal12.1.1a and 12.1.1b. For example, μ-type toxins (M-superfamily, Cys-framework 3) and μO-toxins (O-superfamily, framework 6) preferentially block different isoforms of voltage-gated Na+ channels in nerve and muscle (Terlau and Olivera, 2004; Daly et al., 2004; McIntosh et al., 1995). (A,B) The NSI-IT mass spectra collected for the peptide in the native (A) and reduced–alkylated (B) form. Bull. Despite the high degree of sequence diversity in the toxin-coding region of Cal12.1 cDNAs, there is virtually no variation in the leader and propeptide regions. (B) Toxin-coding regions of Cal12.1.1a and Cal12.2a. Derived from cuttlefish ink, Sepia has a broad range of action over the female organism, and is one of Samuel Hahnemann’s greatest contributions to the homeopathic … Given these differences and the uncertainties in secondary structure, we feel that assignment of cal12a and cal12b peptides (and all the other Cal12.1 and Cal12.2 members) to a specific functional type of conotoxins remains an open question. While children may be lively and excitable, adults are inclined to weariness, indifference, hardness, and irritability. Peptides eluting in the complex region between 30–37 min also had blocking activity against squid Na+ channels, but these effects were essentially irreversible, and fractions in this region were not studied further. 1). Ions were desorbed with a pulsed nitrogen laser (337 nm) and extracted at 20 kV with a 150 ns delay. ... Biol. 3B) shows that the intra-cysteine loops have essentially no amino acid identity and sizeable differences in length. In cuttlefish, the attack includes three components: attention, positioning, and seizure. Packaging Sepia (Sepia) officinalis Linnaeus, 1758 Sepia filliouxi Lafont, 1869 Sepia officinalis mediterranea Ninni, 1884 Sepia rugosa Bowdich, 1822 Sepia vicellius Gray, 1849 Sepia … Other sequences (see Discussion) are: C. vexillum 8 (Kauferstein et al., 2005), PVIA [C. purpurascens (West et al., 2005)], MrVIB [C. marmoreus (McIntosh et al., 1995)]. Diseases (Herbs). Sepia officinalis ingår i släktet Sepia och familjen Sepiidae. A low signal-to-noise ratio prevented clear identification of additional residues. The functional effects of the 6-Cys toxin, Conus vexillum 8, are not known, but this sequence also encodes a framework 6/7 conotoxin (Kauferstein et al., 2005). The peptides cal12a and cal12b correspond to the cDNA sequences Cal12.1.1a and Cal12.1.1b, respectively. We have never identified a clone for any Cal12.1 member in our cDNA library, so sequences corresponding to these amino acids and the 5′ UTR are not available. A diverse family of novel peptide toxins from an unusual cone snail. The Bahia Asuncion snails inhabited the sand beneath a sea-grass bed during low tide and appeared to forage more widely on an adjacent rocky area at high tide. Please log in to add an alert for this article. Inward current that remains at the end of a voltage step (and the inward tail current after the step) flow primarily through Ca+ channels (McFarlane and Gilly, 1996), and these channels are not appreciably affected. Osnovna škola Stobre č 14 ZBIRKA MORSKIH BESKRALJE Ž NJAKA Pu ževi PUZLATKA / PETROVO UHO (Haliotis tuberculata lamellosa) ( Lamarck , 1822.) The cuttlebone is relatively ellipsoid in shape. Breitarm-Sepia (Sepia latimanus) Gewöhnlicher Tintenfisch (Sepia officinalis) Metasepia Hoyle, 1885 Prachtsepia (Metasepia pfefferi) Sepiella Gray, 1849; Sepiadariidae Fischer, 1882 Sepiadarium Steenstrup, 1881; Sepioloidea d’Orbigny, 1845; Idiosepiidae Appelöf, 1898 (von manchen Autoren wird diese Familie auch als Ordnung akzeptiert). They are a migratory species that spend the summer and spring inshore for spawning and then move to depths of 100 to 200m during autumn and winter. Venom from 30 ducts (Monterey snails) was pooled and homogenized in acidified acetone (40:6:1, acetone:water:HCl). A single example of Cal12.1.2h was found in one snail. An active fraction that was eluted for ∼43 min was found to have a doublet peak. site or the information on links from this site to diagnose or treat a health problem
amino acids 9–18, 23–28 and 37–45) and the modular diversity in other regions (amino acids 19–23 and 34–37), it is possible that all isoforms block voltage-gated Na+ channels. In the most extreme case, two Bahia Asuncion snails were each found to have five different Cal12.1 mRNAs with only Cal12.1.1a and Cal12.1.3a in common. Der Zwergtintenfisch-Nachwuchs … Each fraction was individually tested for bioactivity; both peptides reversibly blocked squid Na+ channels at 0 mV with an estimated inhibitor constant (Ki) of 10–20 nmol l–1 (Fig. ‘Milked’ venom was also collected by arousing a snail with a small piece of squid (L. opalescens) skin (internal layer) stretched over a 0.5 ml microfuge tube and enticing a venom injection into the vial (Jakubowski et al., 2005). Effects on Na+ channels in other taxa, including worms and vertebrates, also remain to be determined. We therefore deemed a sequence to be valid only if it could not have been a theoretical recombination product of other sequences found in the same snail. (A) Effect of crude duct venom, diluted 1:100 in the external recording solution. However, whether animals show disruptive patterns is dependent not only on object size but also on their body size. Mass spectra were interpreted manually. J. Linn. It is also known as “African marigold” and has been a subject of several chemical and pharmacological studies. displayed on this site without such supervision, you are solely and entirely responsible
Biol. The number of sequences independently obtained with RT-PCR analysis from an individual snail varied from 19 to 56 (mean ± s.d.=35±9; total number of sequences=1295). HOMŒOPATHIC MATERIA MEDICA. In general, the variable regions form the active surface that defines target specificity. This idea is based on a similar modular structure of scorpion toxins that block voltage-gated potassium (Kv1) channels, with selectivity for specific channel isoforms being dictated by amino-acid side chains in variable regions that are flanked by highly conserved sequences (Ellis et al., 2001; Garcia et al., 2001; Gross and MacKinnon, 1996; Miller, 1995). GenBank accession numbers are given below the name of each Cal12.2 variant. This yielded a fraction with biological activity that produced a single chromatographic peak. Paired fins run from behind the head to the tip of the body. As discussed in the Results, this sequence characterizes the peptides cal12a and cal12b, with position 17 being a modified tryptophan residue. The species undergoes seasonal migrations between inshore waters during spring and summer and medium shelf grounds (about 100 m depth) during autumn and winter. 10A). Samudraphena (Internal-cell of Sepia officinalis. Amplifications from individual snails also yielded sequences (in ∼25% of the animals) that differed from Cal12.1.1a at only two nucleotides in the toxin-coding region. Unioonpeedia on mõiste kaardi või semantiline võrgustik, korraldati nagu entsüklopeedia - sõnastik. does not including the head and the arms), but most are smaller, around 20 to 30 cm. Services advises you to always seek the advice of your physician or other qualified
We excluded additional sequences based on two major concerns – potential DNA polymerase errors and recombination of mixed template species during PCR and cloning. Alternatively, if C2 and C6 of the Cal12.1-encoded peptide were linked as depicted in Fig. Flow from the microbore HPLC was directly sprayed into the LCQ Deca IT mass spectrometer through the electrospray ionization (ESI) source with a spray voltage of 4.3 kV and nebulizing gas flow of 40 units, facilitating desorption. Some of the more intense parent masses are labeled based on the collected mass spectra from each run, and different snails clearly express different peptides. Marine cone snails inject prey with paralytic peptide toxins, and more than 500 extant Conus species produce an enormous number of unique Cys-rich conotoxins (Gray et al., 1988; Norton and Olivera, 2006; Rockel et al., 1995). In four snails it was the only transcript found (19–35 sequences), but in three snails it was not detected at all (22–48 clones sequenced). Testing gastropods that were actually preyed on by C. californicus would be valuable, as would the testing of Ca2+ and K+ currents in ecologically relevant gastropods. 2D), and this subfraction appeared to contain a single peptide species (Fig. By contrast, this study examines Conus californicus, which is distantly related to the Indo-Pacific species (Duda and Kohn, 2005; Duda et al., 2001; Espiritu et al., 2001). Using molecular cloning of cDNAs and mass spectrometry, we examined peptides isolated from venom ducts to elucidate the sequences and post-translational modifications of two eight-cysteine toxins (cal12a and cal12b of type 12 framework) that block voltage-gated Na+ channels. Prominent catches of unidentified cuttlefishes (Sepia spp. Leader and propeptide sequences of O-type peptides (C-C-CC-C-C framework) from other Conus species are also shown, and identities with Cal12.1.1a are indicated by the light gray shading. Animal Cognition 9 2006. If you undertake any treatment methods
All organic solvents used in MS were of high performance liquid chromatography (HPLC) grade or better (Thermo Fisher Scientific, Waltham, MA, USA). These variants are therefore included in this report. Paired fins run from behind the head to the tip of the body. Conus canonicus). One substitution (GAA to GAT) leads to a conservative change of Glu to Asp at position 21, but the second substitution (GGT to GGC) is synonymous. Over 100 specimens were collected. Sepia sp. The solvent gradient was 50 μl min–1 from 20–25% B for the first 5 min, 25–40% B for 60 min, and then ramped up to 80% B to flush the column of any remaining compounds. PCR products were purified and sequenced using the nested T3/T7 primers (Table 1). Note the increased fragmentation after the disulfide bonds are broken. (D) Effect of purified subfraction 8 peptide at a concentration of ∼25 nmol l–1. vol. No 5′ UTR sequence is available. The sample (5–10 μl) was then frozen at –80°C. Fins are starting directly at the anterior edge of the mantle and they are extending beyond the edge noticeably, reaching anteriorly to level of posterior edge of eyes; fins widen at posterior part of the mantle. Moreover, five of the 15 Cal12.1 variants were found only in Bahia Asuncion snails. A high degree of diversity clearly exists in the Cal12.1 family of cDNAs, but the 15 different variants reported in this paper represent a conservative estimate. Single-duct venom liquid chromatography–mass spectrometry Later purifications focused on venom isolated from single ducts (Monterey snails). (B) Amino acid alignments of the toxin-coding regions of known peptides with the 8-Cys, type-12 framework. Three additional Cal12.2 variants were also found in the cDNA library. The region of amino acids 19–22 allows division into subfamilies (Cal12.1–1.3), with each subfamily having a ‘signature’ consensus sequence indicated by the outlined boxes. Calendula officinalis Linn. and by award no. Biol. Although such a diverse diet suggests that C. californicus may express novel toxins, this species has been largely overlooked (Cottrell and Twarog, 1972; Elliott and Kehoe, 1978; Elliott and Raftery, 1979; Whysner and Saunders, 1963; Whysner and Saunders, 1966).