Asterias Rubens Research Articles

Isolation and characterization of NGFFYamide neuropeptide from Patiria pectinifera pyloric caeca extract

Neuropeptides are small molecules that mediate intercellular signaling and regulate physiological processes. Starfish possess various myoactive neuropeptides, including starfish myorelaxant peptide (SMP) and a calcitonin-type peptide with apical muscle relaxing properties. In this study, we report the purification of a neuropeptide from starfish (Patiria pectinifera) pyloric caeca extract using high-performance liquid chromatography (HPLC) and an in vitro bioassay to screen for fractions and peptides with relaxing effects on P. pectinifera apical muscle preparations. A series of HPLC steps using reversed-phase and cation-exchange columns yielded a purified peptide with muscle-relaxing effects. The purified peptide's structure was determined by LC-MS and Edman degradation, revealing a pentapeptide with an amidated C-terminus (NGFFYamide) and a molecular mass of 646.2930 Da. This is the first report of NGFFYamide purification from P. pectinifera through biochemical methods. The nucleotide sequence encoding the NGFFYamide precursor was determined, showing the presence of a conserved neurophysin domain in the C-terminal region. RT-qPCR results confirmed high expression in radial nerves cord, consistent with previous findings on NG peptides in echinoderms. In vitro pharmacological studies on muscle preparations from P. pectinifera and Asterias amurensis revealed differential relaxing activity of NGFFYamide on apical muscles, while its effects on tube foot preparations were similar in both species. NGFFYamide also induced potent contraction in P. pectinifera cardiac stomach. Comparison of three NG peptides (NGFFYamide, NGFFFamide, and NGIWYamide) on P. pectinifera cardiac stomach revealed varying potency, suggesting class-specific receptor interactions. Tachyphylaxis was observed in P. pectinifera apical muscle but not in A. amurensis, warranting further investigation. Based on these results, it is plausible that NGFFYamide could be involved in regulating locomotion and feeding behavior in P. pectinifera, consistent with findings in Asterias rubens.

Evaluation of Anti-Cancer Potential of Homeopathic Medicines by In-Vitro Study on Human Breast, Lung and Colorectal Cancer Cells

The purpose of this study is to assess the cytotoxic effects of homoeopathic drugs on cancer cell lines by In-vitro experiment. Background: Homoeopathy is a renowned therapeutic approach in many nations across the world. However, the approach has sparked much scientific discussion concerning its efficacy and methods of action. This is particularly true when it comes to the application of homeopathy in cancer treatment. Objective: The purpose of this study was to evaluate the cytotoxic effect of homoeopathic remedies on cancer cells and to emphasise the therapeutic effects of 50 Millesimal potencies. Materials & Methods: Carcinosinum, Conium maculatum, and Asterias rubens were the drugs used in 0/1 potency against the MCF 7 breast cancer cell line. Carcinosinum, Nitric acid, and Ruta were the drugs in 0/1 potency used against the HCT 116 colorectal cancer cell line and the NCI-H460 lung cancer cell line. 90% alcohol medicated on poppy-sized globules, dried fully and used as a control for all cell lines. The cytotoxicity has been assessed using the standard MTT test. Results: The LC50 values of the medicines tested on breast and lung cancer cells were not obtained up to 40000 ppm concentrations, as a result, cytotoxicity evaluation was not possible. The LC50 values of the medicines tested on Colo-rectal cancer cells were obtained around 20,000 ppm concentration and showed a gradual increase in Cell Death with increasing Concentration of exposure of the drugs. As a result, these drugs revealed cytotoxic qualities as well as anti-cancer actions. Conclusion: Despite the objective facts, we cannot reach a definite conclusion about the anti-cancer effectiveness of these drugs in vitro because several aspects of homeopathy's action are unlikely to be replicated in In-vitro study. Determination of reasons for the same or understanding the molecular mechanism of resistance of the cell lines to the cytotoxic effects of the drugs were not within the purview of the study and these can be addressed in future studies. Keywords: Homeopathy, Cancer, MCF-7, HCT 116, NCI-H 460, In-Vitro, Carcinosinum, Conium maculatum, Asteria rubens, Ruta graveolens, Nitric acid, 50 Millesimal Potency, MTT Assay.

An evaluation of acoustic telemetry as a method to study the movements of Asteroidea (Asterias rubens)

BackgroundAcoustic telemetry is an important tool to study the movement of aquatic animals. However, studies have focussed on particular groups of easily tagged species. The development of effective tagging methods for ecologically important benthic species, such as sea stars, remains a challenge due to autotomy and their remarkable capacity to expel any foreign material. We tested three methods to surgically attach acoustic transmitters to the common sea star Asterias rubens; two methods attached the tag to the aboral side of the central body and the third attached the transmitter to the aboral side of an arm. Laboratory experiments evaluated each method in terms of survivability, tag retention, associated injuries, and changes in feeding behaviour and physical condition.ResultsLaboratory results were highly variable; however, all tagging methods caused significant injury to the epidermis and deeper tissue around the attachment site over periods greater than 4 weeks. Attaching a tag by horizontally piercing the central body (method HPC) had minimal effects in the short-term (2–3 weeks) and this method was used for a pilot tagging study in the field, where 10 sea stars were tagged and placed within an existing acoustic telemetry array. Although, the interpretation of field data was challenging due to the characteristic slow movement of sea stars, movements of a similar magnitude to previous studies were identified during the 2–4 weeks after sea stars were tagged and released. However, this apparent period of tagging success was followed by a reduction in movement that, when viewed in conjunction with laboratory results, potentially indicated a deterioration in the sea stars’ physical condition.ConclusionsWhile acoustic telemetry continues to provide novel insights into the ecology of a wide variety of marine species, species-specific effects of tagging should be evaluated before starting field studies. If the autonomous study of benthic movement is to expand beyond hard-bodied macroinvertebrates current methodological and analytical challenges must be addressed.

High-quality chromosome-level genome assembly of the Northern Pacific sea star Asterias amurensis.

Asterias amurensis, a starfish species that is native to countries such as China and Japan, as well as non-native regions like Australia, has raised serious concerns in terms of its impact on ecology and economy. To gain a better understanding of its population genomics and dynamics, we successfully assembled a high-quality chromosome-level genome of A. amurensis using PacBio and Hi-C sequencing technologies. A total of 87 scaffolds assembly with contig N50 length of 10.85Mb and scaffold N50 length of 23.34Mb were obtained, with over 98.80% (0.48 Gb) of them anchored to 22 pseudochromosomes. We predicted 16,673 protein-coding genes, 95.19% of which were functionally annotated. Our phylogenetic analysis revealed that A. amurensis and Asterias rubens formed a clade, and their divergence time was estimated ~ 28 million years ago (Mya). The significantly enriched pathways and Gene Ontology terms related to the amplified gene family were mainly associated with immune response and energy metabolism, suggesting that these factors might have contributed to the adaptability of A. amurensis to its environment. This study provides valuable genomic resources for comprehending the genetics, dynamics, and evolution of A. amurensis, especially when population outbreaks or invasions occur.

An open‐source general purpose machine learning framework for individual animal re‐identification using few‐shot learning

Abstract Animal re‐identification remains a challenging problem due to the cost of tagging systems and the difficulty of permanently attaching a physical marker to some animals, such as sea stars. Due to these challenges, photo identification is a good fit to solve this problem whether evaluated by humans or through machine learning. Accurate machine learning methods are an improvement over manual identification as they are capable of evaluating a large number of images automatically and recent advances have reduced the need for large training datasets. This study aimed to create an accurate, robust, general purpose machine learning framework for individual animal re‐identification using images both from publicly available data as well as two groups of sea stars of different species under human care. Open‐source code was provided to accelerate work in this space. Images of two species of sea star (Asterias rubens and Anthenea australiae) were taken using a consumer‐grade smartphone camera and used as original datasets to train a machine learning model to re‐identify an individual animal using few examples. The model's performance was evaluated on these original sea star datasets which contained between 39–54 individuals and 983–1204 images, as well as using six publicly available re‐identification datasets for tigers, beef cattle noses, chimpanzee faces, zebras, giraffes and ringed seals ranging between 45–2056 individuals and 829–6770 images. Using time aware‐splits, which are a data splitting technique ensuring that the model only sees an individual's images from a previous collection event during training to avoid information leaking, the model achieved high (>99%) individual re‐identification mean average precision for the top prediction (mAP@1) for the two species of sea stars. The re‐identification mAP@1 for the mammalian datasets was more variable, ranging from 83% to >99%. However, this model outperformed published state‐of‐the‐art re‐identification results for the publicly available datasets. The reported approach for animal re‐identification is generalizable, with the same machine learning framework achieving good performance in two distinct species of sea stars with different physical attributes, as well as seven different mammalian species. This demonstrates that this methodology can be applied to nearly any species where individual re‐identification is required. This study presents a precise, practical, non‐invasive approach to animal re‐identification using only basic image collection methods.

Supercritical CO2 for efficient extraction of high-quality starfish (Asterias rubens) oil

The growing demand for healthy marine lipids and the unmatched supply demand required diversification of their sources. Characterization of starfish oil has shown that it is a new possible source of marine lipids. However, optimized extraction conditions are needed to extract high quality oil. Hence, this study aimed to extract starfish oil using supercritical carbon dioxide (ScCO2) and optimize the extraction process using response surface methodology (RSM). Temperature, pressure, and ethanol flow rate were the most influential parameters for oil extraction. The RSM model predicted 19.40% and 25.40% for extraction yield and phospholipids content respectively at 40 °C, 350 bar, and 2.61 mL/min of ethanol flow rate. The result of validation extraction showed close agreement with the model. Moreover, the oils were characterized by fatty acid composition, peroxide value, free fatty acid, and tocopherol contents. The current study showed that high-quality lipids could be extracted from underutilized species using ScCO2 extraction.

Asterias rubens in High Dilutions exhibiting Anti-Cancer Activity in Breast Cancer Cell Line: An in vitro Study

Asterias rubens constitutes saponins that represents a class of steroid oligoglycoside namely asterosaponins that has broad biological functions inclusive of anti-cancerous activity. The most common malignancy as well as the second largest cause of mortality related to cancer in women is breast carcinoma. The prime objective of this research is to assess the anti-cancer activity of Asterias rubens in its higher dilutions on MCF-7 cells and to evaluate the process of angiogenesis. In vitro cytotoxicity was assessed by MTT and SRB assay. To evaluate the long-term survival capacity and proliferative ability of MCF-7 cell line, colony forming assay was performed. CAM assay was performed to evaluate the angiogenic potential of Asterias rubens. It was observed that Asterias rubens showed significant anti-cancer activity. In the in vivo study, chick embryo treated with Asterias rubens showed angiogenesis. Asterias rubens in higher dilutions exhibited anti-cancer activity, signifying its utility as supportive medicine in cancer treatment.

Evidence of introgression, ecological divergence and adaptation in Asterias sea stars.

Hybrid zones are important windows into the evolutionary dynamics of populations, revealing how processes like introgression and adaptation structure population genomic variation. Importantly, they are useful for understanding speciation and how species respond to their environments. Here, we investigate two closely related sea star species, Asterias rubens and A. forbesi, distributed along rocky European and North American coastlines of the North Atlantic, and use genome-wide molecular markers to infer the distribution of genomic variation within and between species in this group. Using genomic data and environmental niche modelling, we document hybridization occurring between northern New England and the southern Canadian Maritimes. We investigate the factors that maintain this hybrid zone, as well as the environmental variables that putatively drive selection within and between species. We find that the two species differ in their environmental niche breadth; Asterias forbesi displays a relatively narrow environmental niche while conversely, A. rubens has a wider niche breadth. Species distribution models accurately predict hybrids to occur within environmental niche overlap, thereby suggesting environmental selection plays an important role in the maintenance of the hybrid zone. Our results imply that the distribution of genomic variation in North Atlantic sea stars is influenced by the environment, which will be crucial to consider as the climate changes.

Gonadotropic activity of a second relaxin-type peptide in starfish

In starfish, a relaxin-like gonad-stimulating peptide (RGP) acts as a gonadotropin that triggers gamete maturation and spawning. In common with other relaxin/insulin superfamily peptides, RGP consists of an A- and a B-chain, with cross-linkages mediated by one intra- and two inter-chain disulfide bonds. In this study, a second relaxin-like peptide (RLP2) was identified in starfish species belonging to the orders Valvatida, Paxillosida, and Forcipulatida. Like RGP, RLP2 precursors comprise a signal peptide and a C-peptide in addition to the A- and B-chains. However, a unique cysteine motif [CC-(3X)-C-(10X)-C] is present in the A-chain of RLP2, which contrasts with the cysteine motif in other members of the relaxin/insulin superfamily [CC-(3X)-C-(8X)-C]. Importantly, in vitro pharmacological tests revealed that Patiria pectinifera RLP2 (Ppe-RLP2) and Asterias rubens RLP2 (Aru-RLP2) trigger shedding of mature eggs from ovaries of P. pectinifera and A. rubens, respectively. Furthermore, the potencies of Ppe-RLP2 and Aru-RLP2 as gonadotropic peptides were similar to those of Ppe-RGP and Aru-RGP, respectively, and the effect of RLP2 exhibited partial species-specificity. These findings indicate that two relaxin-type peptides regulate spawning in starfish and therefore we propose that RGP and RLP2 are renamed RGP1 and RGP2, respectively.

Effect of electromagnetic fields from renewable energy subsea power cables on righting reflex and physiological response of coastal invertebrates

Offshore renewables are expanding, yet more information is required to understand their possible impacts on the environment. Little is known about the effects of Electromagnetic Fields (EMF) from subsea power cables on marine life. This study simulated an EMF of 500 μT, as modelled for an export cable over a rocky shore, where the industry standard cable burial would not be possible. Righting reflex, refractive index of haemolymph/coelomic fluid, and total haemocyte/coelomocyte counts were measured for four coastal invertebrates (Asterias rubens, Echinus esculentus, Necora puber, and Littorina littorea). No significant differences were found in either behavioural or physiological responses. This was the first study to investigate EMF exposure on righting reflex, and the first ever EMF study on edible sea urchins and periwinkles, and only one of a couple for common starfish and velvet crabs. It therefore, provides valuable data for environmental impact assessments, marine spatial planning, and commercial fisheries.

Structural and functional properties of collagen isolated from lumpfish and starfish using isoelectric precipitation vs salting out.

The possibility of replacing the very time and resource demanding salting out (SO) method with isoelectric precipitation (IP) during collagen extraction from common starfish and lumpfish was investigated. The effect of IP on yield, structural and functional properties of the collagens was therefore compared with SO. Application of IP resulted in a higher or similar collagen mass yield compared with SO from starfish and lumpfish, respectively. However, the purity of collagens recovered with IP was lower than those recovered with SO. Replacing SO with IP did not affect polypeptide pattern and tropohelical structural integrity of collagen from the two resources as revealed with SDS-PAGE and FTIR analysis. Thermal stability and fibril formation capacity of collagens recovered with IP were also well preserved. Overall, the results showed that the IP can be a promising resource smart alternative for the classic SO precipitation during collagen extraction from marine resources.

Localization of relaxin-like gonad-stimulating peptide expression in starfish reveals the gonoducts as a source for its role as a regulator of spawning.

Oocyte maturation and gamete release (spawning) in starfish are triggered by relaxin-like gonad-stimulating peptide (RGP), a neuropeptide that was first isolated from the radial nerve cords of these animals. Hitherto, it has generally been assumed that the radial nerve cords are the source of RGP that triggers spawning physiologically. To investigate other sources of RGP, here we report the first comprehensive anatomical analysis of its expression, using both in situ hybridization and immunohistochemistry to map RGP precursor transcripts and RGP, respectively, in the starfish Asterias rubens. Cells expressing RGP precursor transcripts were revealed in the ectoneural epithelium of the radial nerve cords and circumoral nerve ring, arm tips, tube feet, cardiac stomach, pyloric stomach, and, most notably, gonoducts. Using specific antibodies to A. rubens RGP, immunostaining was revealed in cells and/or fibers in the ectoneural region of the radial nerve cords and circumoral nerve ring, tube feet, terminal tentacle and other arm tip-associated structures, body wall, peristomial membrane, esophagus, cardiac stomach, pyloric stomach, pyloric caeca, and gonoducts. Our discovery that RGP is expressed in the gonoducts of A. rubens proximal to its gonadotropic site of action in the gonads is important because it provides a new perspective on how RGP may act as a gonadotropin in starfish. Thus, we hypothesize that it is the release of RGP from the gonoducts that triggers gamete maturation and spawning in starfish, while RGP produced in other parts of the body may regulate other physiological/behavioral processes.

Sea Star Wasting Disease in captive common sea stars (Asterias rubens): Examinations, therapy attempts and course of disease

Sea Star Wasting Disease (SSWD) affects several wild and captive starfish species. It is generally thought to be infectious, with environmental conditions favouring its progression. Captive Asterias rubens with varying degrees of epidermal ulceration, limb autotomy and eversion of viscera were examined. Antibiotic treatments by injections and baths did not reduce mortality.

Nonconsumptive Effects of Crustaceans and an Echinoderm on Spat of the Eastern Oyster Crassostrea virginica (Gmelin, 1791)

Predation by crustaceans and echinoderms is a significant driver of mortality for sessile invertebrates in near-shore marine ecosystems. As a response, shellfish growers use predator exclusion cages to grow their product. Oysters grown in these cages and clams in and under nets have been observed to exhibit thinner, brittle shells, which is not optimal for market, nor for those being grown for restoration purposes. Here, the nonconsumptive predator effects of several native and nonnative predators on shell morphology of Crassostrea virginica oyster spat. Juvenile (1–3 mm) seed oysters were exposed separately to chemical cues of five different predators and one nonpredator crab were investigated and compared with control (no exposure) spat. Significant effects of predator presence on the total wet weight (g) of the spat was demonstrated, with the highest increase for spat exposed to black-fingered mud crabs (Panopeus, 1.70 mg d–1) and the biggest decreases for spat exposed to the European green crabs (Carcinus maenas—1.34 mg d–1) and the sea stars (Asterias rubens—1.47 mg d–1). Changes to oyster length (mm) and shell density (crushing resistance) were not significant. Implications for aquaculturists and wild bivalve populations are that growth could be negatively affected by the lack of predators on these important organisms.

Predators regulate the taxonomic structure of mixed Mytilus edulis L. and M.trossulus Gould settlements in the shallow waters of the White Sea

Cryptic mussel species Mytilus edulis (Me) and M.trossulus (Mt) form mixed settlements in shallow waters of the White Sea. The proportion of Mt and Me in local settlements varies in broad limits but the factors regulating taxonomic composition of mixed populations are poorly understood. Present study focuses on the assessment of the sea star (Asterias rubens) predation influence on the Mt-Me ratio in natural habitats. For this purpose, we’ve conducted a series of field experiments. For mussels placed on experimental units (ceramic plates submerged in starfish-infested habitats), we’ve estimated a probability of being eaten by starfish. We’ve constructed a linear model that includes several predictors which may influence the probability of being eaten (starfish biomass, mussel size, mussel species, prey abundance, Mt-Me ratio on experimental units and abundance of conspecifics). The analysis has revealed that the probability of being eaten expectedly increases with sea stars’ biomass increasing, but it is higher for Mt in comparison with Me. This indicates Mt as a preferable prey. Negative correlations between the probability of being eaten and shell size, number of conspecifics and mussel abundance were also revealed. No significant dependence on Mt-Me ratio was found. Additionally, we’ve taken field samples which revealed a significant decrease in the proportion of Mt in mussel patches after a massive starfish attack. This indicates that starfish ate out Mt as its preferable prey. These results show that predators can contribute to the regulation of taxonomic composition of mixed Mt-Me settlements in the White Sea.

Morphology and Settling Velocity of Sea Stars (Asterias rubens)

This study presents morphological data and estimates of the settling velocity and drag coefficient of sea stars (Asterias rubens) caught in the Limfjord, Denmark. A geometric model describing the sea star is presented and the thickness and arm width are determined as linear functions of arm length. The volume and mass of the sea stars is well predicted by the geometric model and is in agreement with the experimental measurements. The mean sea star density is determined to be 1095 kg/m3, the mean drag coefficient is estimated to be 2.3 and the settling velocity is shown to vary with the square root of the its size.

Immunohistochemical localisation of vasopressin/oxytocin-type, corazonin-type and luqin-type neuropeptide expression in the starfish Asterias rubens using antibodies to the C-terminal region of precursor proteins

Neuropeptides derived from larger precursor proteins are secreted as signalling molecules by neurons and regulate diverse physiological and behavioural processes in animals. Recently, we reported the discovery of ArCRZ (HNTFTMGGQNRWKAG-NH2) and ArLQ (EEKTRFPKFMRW-NH2)—novel neuropeptides in the starfish Asterias rubens that are orthologs of arthropod corazonins and molluscan luqins, respectively. However, our efforts to generate antibodies to ArCRZ and ArLQ have been unsuccessful, precluding immunohistochemical analysis of their expression. Here, we investigated an alternative experimental approach for neuropeptide immunohistochemistry by generating antibodies to peptides corresponding to the C-terminal region of the precursor proteins. As proof of principle, we generated antibodies to the C-terminal region of the precursor of the vasopressin/oxytocin-type neuropeptide asterotocin and show that these reveal immunostaining in A. rubens that is very similar to that observed with asterotocin antibodies. Furthermore, antibodies to the C-terminal region of the ArCRZ precursor (ArCRZP) and the ArLQ precursor (ArLQP) produced patterns of immunostaining consistent, respectively, with the distribution of ArCRZP and ArLQP transcripts revealed by mRNA in situ hybridisation. Detailed immunohistochemical analysis revealed widespread expression of ArCRZP and ArLQP in A. rubens, including the central nervous system, digestive system and the body wall and its associated appendages (e.g. tube feet), providing a neuroanatomical framework for investigation and interpretation of the pharmacological actions of ArCRZ and ArLQ in A. rubens. Furthermore, our findings provide a basis for use of antibodies to the C-terminal region of neuropeptide precursor proteins in other species where the production of antibodies to the bioactive neuropeptides is unsuccessful.

Common Sea Star (Asterias rubens) Coelomic Fluid Changes in Response to Short-Term Exposure to Environmental Stressors

Common sea stars (Asterias rubens) are at risk of physiological stress and decline with projected shifts in oceanic conditions. This study assessed changes in coelomic fluid (CF) blood gases, electrolytes, osmolality, and coelomocyte counts in adult common sea stars after exposure to stressors mimicking effects from climate change for 14 days, including decreased pH (−0.4 units, mean: 7.37), hypoxia (target dissolved oxygen ~1.75 mg O2/L, mean: 1.80 mg O2/L), or increased temperature (+10 °C, mean: 17.2 °C) and compared sea star CF electrolytes and osmolality to tank water. Changes in CF blood gases, electrolytes, and/or coelomocyte counts occurred in all treatment groups after stressor exposures, indicating adverse systemic effects with evidence of increased energy expenditure, respiratory or metabolic derangements, and immunosuppression or inflammation. At baseline, CF potassium and osmolality of all groups combined were significantly higher than tank water, and, after exposures, CF potassium was significantly higher in the hypoxia group as compared to tank water. These findings indicate physiological challenges for A. rubens after stressor exposures and, given increased observations of sea star wasting events globally, this provides evidence that sea stars as a broad group are particularly vulnerable to changing oceans.

Elevated temperature linked to signs associated with sea star wasting disease in a keystone European species, Asterias rubens

Sea star wasting disease (SSWD) refers to a suite of gross signs affecting Asteroidea species. These include epidermal lesions, everted viscera, arm autotomy, and ultimately, full body disintegration leading to mortality. The common sea star Asterias rubens is a keystone species in the coastal Northeast Atlantic and may be susceptible to the disease. While the precise aetiology of SSWD remains poorly understood, environmental instability, including rising sea temperatures, has been linked to SSWD outbreaks. To investigate this connection, an experiment was conducted to quantify disease sign expression in A. rubens under elevated temperature. We exposed sea stars to either elevated temperature (18°C) or a control treatment (12°C) for a 14 d period. We quantified the presence of disease signs associated with SSWD, the progression of signs, and survival of individuals. Elevated temperature induced a greater number of signs consistent with SSWD and also resulted in mortality for some of those animals. Furthermore, larger individuals were more likely to show increased signs of disease. Our results provide evidence that signs associated with SSWD increase with elevated temperature.

Starfish smooth muscle relaxing activity of SALMFamide isotype peptide and its analog derived from starfish, Asterias rubens

An organism’s physiological processes and behaviors are regulated by neuropeptides and hormone peptides. The first neuropeptide identified from echinoderms is SALMFamide. The two most well-studied SALMFamide neuropeptides are S1 and S2, which possess myoactivity on apical muscle, tube feet, and the cardiac stomach of starfishes. However, neuropeptide candidates identified from SALMFamide’s precursor protein sequence have not been investigated. This study aims to compare the bioactivity of SALMFamide neuropeptides from the starfish Asterias rubens using various starfish muscle preparations. In this study, the bioactivity of the L-type SALMFamide neuropeptides from the starfish A. rubens, AYHTGLPFamide (SALMFa-A) and the derivative AYHSALMFamide (SALMFa-B) was investigated. The neuropeptides were applied on Asterias amurensis apical muscle, tube feet, which revealed that the neuropeptides exhibit relaxing activity on apical muscle but no activity on tube feet. The native SALMFa- A peptide had lower relaxing activity on the apical muscle compared to the derivative peptide SALMFa-B. The relaxing activity of two neuropeptides also was compared with those on the apical muscle of Patiria pectinifera, which revealed relaxing activity as well as SALMFamide-S1 and S2 neuropeptides. Moreover, the investigation of SALMFa-A and SALMFa-B peptides’ bioactivity on P. pectinifera cardiac stomach muscle also showed slight relaxing activity.