The branched mitochondrial respiratory chain from the jellyfish Stomolophus sp2 as a probable adaptive response to environmental changes.

During their long evolutionary history, jellyfish have faced changes in multiple environmental factors, to which they may selectively fix adaptations, allowing some species to survive and inhabit diverse environments. Previous findings have confirmed the jellyfish's ability to synthesize large ATP amounts, mainly produced by mitochondria, in response to environmental challenges. This study characterized the respiratory chain from the mitochondria of the jellyfish Stomolophus sp2 (previously misidentified as Stomolophus meleagris). The in-gel activity from isolated jellyfish mitochondria confirmed that the mitochondrial respiratory chain contains the four canonical complexes I to IV and F0F1-ATP synthase. Specific additional activity bands, immunodetection, and mass spectrometry identification confirmed the occurrence of four alternative enzymes integrated into a branched mitochondrial respiratory chain of Stomolophus sp2: an alternative oxidase and three dehydrogenases (two NADH type II enzymes and a mitochondrial glycerol-3-phosphate dehydrogenase). The analysis of each transcript sequence, their phylogenetic relationships, and each protein's predicted models confirmed the mitochondrial alternative enzymes' identity and specific characteristics. Although no statistical differences were found among the mean values of transcript abundance of each enzyme in the transcriptomes of jellyfish exposed to three different temperatures, it was confirmed that each gene was expressed at all tested conditions. These first-time reported enzymes in cnidarians suggest the adaptative ability of jellyfish's mitochondria to display rapid metabolic responses, as previously described, to maintain energetic homeostasis and face temperature variations due to climate change.

Digestive glycosidases from cannonball jellyfish (Stomolophus sp. 2): identification and temporal-spatial variability.

Jellyfish are economically important organisms in diverse countries, carnivorous organisms that consume various prey (crustaceans, mollusks, bivalves, etc.) and dissolved carbohydrates in marine waters. This study was focused on detecting and quantifying the activity of digestive glycosidases from the cannonball jellyfish (Stomolophus sp. 2) to understand carbohydrate digestion and its temporal-spatial variation. Twenty-three jellyfish gastric pouches were collected in 2015 and 2016 in the Gulf of California in three localities (Las Guásimas, Hermosillo, and Caborca). Nine samples were in intra-localities from Las Guásimas. Chitinase (Ch), ß-glucosidase (ß-glu), and ß-N-acetylhexosaminidase (ß-NAHA) were detected in the gastric pouches. However, cellulase, exoglucanase, α-amylase, polygalacturonase, xylanase, and κ-carrageenase were undetected. Detected enzymes showed halotolerant glycolytic activity (i = 0-4 M NaCl), optimal pH, and temperature at 5.0 and 30-50 °C, respectively. At least five ß-glucosidase and two ß-N-acetylhexosaminidase were detected using zymograms; however, the number of proteins with chitinase activity is not precise. The annual variation of cannonball jellyfish digestive glycosidases from Las Guásimas between 2015-2016 does not show significant differences despite the difference in phytoplankton measured as chlorophyll α (1.9 and 3.4 mg/m3, respectively). In the inter-localities, the glycosidase activity was statistically different in all localities, except for ß-N-acetylhexosaminidase activity between Caborca and Hermosillo (3,009.08 ± 87.95 and 3,101.81 ± 281.11 mU/g of the gastric pouch, respectively), with chlorophyll α concentrations of 2.6, 3.4 mg/m3, respectively. For intra-localities, the glycosidase activity did not show significant differences, with a mean chlorophyll α of 1.3 ± 0.1 mg/m3. These results suggest that digestive glycosidases from Stomolophus sp. 2 can hydrolyze several carbohydrates that may belong to their prey or carbohydrates dissolved in marine waters, with salinity over ≥ 0.6 M NaCl and diverse temperature (4-80 °C) conditions. Also, chlorophyll α is related to glycosidase activity in both seasons and inter-localities, except for chitinase activity in an intra-locality (Las Guásimas).

Catostylus tagi: partial rDNA sequencing and characterisation of nematocyte structures using two improvements in jellyfish sample preparation

Background More than 200 Scyphozoa species have been described, but few have been properly studied regarding their chemical and genetic characteristics.Catostylus tagi, an edible Scyphozoa and the sole European Catostylidae, occurs in summer at Tagus and Sado estuaries. Neither a systematic comparison between the two Catostyluscommunities nor a chemical approach on their nematocytes had been carried out yet.Methods In order to achieve these purposes, optimisation of DNA extraction and of histochemical staining procedures were developed.Catostylus specimens from Tagus and Sado estuaries were compared by ribosomal 18S, 28S, and ITS1 partial sequencing. The morphochemistry of nematocytes was studied by optical and electronic microscopy.Results Macroscopic and molecular results indicated that both communities belong to the same species, C. tagi. The hematoxylin and eosin staining allowed the visualisation of nematocyst genesis and indicated a basic character for the macromolecules on the shaft of euryteles and on the tubule of isorhizae and birhopaloids. By Massons trichrome procedure, the basic properties of the tubules were confirmed and a collagenous profile for the toxins was suggested. Results of the alcian blue staining showed that the outer membrane of nematocyte may consist of macromolecules with acidic polysaccharides, consistent with NOWA and nematogalectin glycoproteins detected in Hydra, but also with poly-gamma-glutamate complex, chitin-like polysaccharides and hyaluronic acids. Through the von Kossa assays, calcium was detected; its position suggested interactions with polysaccharides of the membrane, with proteins of the contractile system or with both.Conclusions The optimisation of sample preparation for DNA extraction may facilitate further studies on little known jellyfish species. The improvement of the smear procedure simplified the use of stained reactions in zooplankton. Moreover, it was shown that good slide images might be acquired manually. The development of specific reactions, with traditional dyes and others, can give important contributions to clarify the chemical nature of the components of nematocytes. The characterisation of nematocyst toxins by staining tests is a goal to achieve.(AU)

Catostylus tagi: partial rDNA sequencing and characterisation of nematocyte structures using two improvements in jellyfish sample preparation

Background More than 200 Scyphozoa species have been described, but few have been properly studied regarding their chemical and genetic characteristics.Catostylus tagi, an edible Scyphozoa and the sole European Catostylidae, occurs in summer at Tagus and Sado estuaries. Neither a systematic comparison between the two Catostyluscommunities nor a chemical approach on their nematocytes had been carried out yet.Methods In order to achieve these purposes, optimisation of DNA extraction and of histochemical staining procedures were developed.Catostylus specimens from Tagus and Sado estuaries were compared by ribosomal 18S, 28S, and ITS1 partial sequencing. The morphochemistry of nematocytes was studied by optical and electronic microscopy.Results Macroscopic and molecular results indicated that both communities belong to the same species, C. tagi. The hematoxylin and eosin staining allowed the visualisation of nematocyst genesis and indicated a basic character for the macromolecules on the shaft of euryteles and on the tubule of isorhizae and birhopaloids. By Massons trichrome procedure, the basic properties of the tubules were confirmed and a collagenous profile for the toxins was suggested. Results of the alcian blue staining showed that the outer membrane of nematocyte may consist of macromolecules with acidic polysaccharides, consistent with NOWA and nematogalectin glycoproteins detected in Hydra, but also with poly-gamma-glutamate complex, chitin-like polysaccharides and hyaluronic acids. Through the von Kossa assays, calcium was detected; its position suggested interactions with polysaccharides of the membrane, with proteins of the contractile system or with both.Conclusions The optimisation of sample preparation for DNA extraction may facilitate further studies on little known jellyfish species. The improvement of the smear procedure simplified the use of stained reactions in zooplankton. Moreover, it was shown that good slide images might be acquired manually. The development of specific reactions, with traditional dyes and others, can give important contributions to clarify the chemical nature of the components of nematocytes. The characterisation of nematocyst toxins by staining tests is a goal to achieve.

The extract of the jellyfish Phyllorhiza punctata promotes neurotoxic effects.

Phyllorhiza punctata (P. punctata) is a jellyfish native to the southwestern Pacific. Herewith we present the biochemical and pharmacological characterization of an extract of the tentacles of P. punctata. The tentacles were subjected to three freeze-thaw cycles, homogenized, ultrafiltered, precipitated, centrifuged and lyophilized to obtain a crude extract (PHY-N). Paralytic shellfish poisoning compounds such as saxitoxin, gonyautoxin-4, tetrodotoxin and brevetoxin-2, as well as several secretory phospholipase A(2) were identified. PHY-N was tested on autonomic and somatic neuromuscular preparations. In mouse vas deferens, PHY-N induced phasic contractions that reached a peak of 234 ± 34.7% of control twitch height, which were blocked with either 100 µ m of phentolamine or 1 m m of lidocaine. In mouse corpora cavernosa, PHY-N evoked a relaxation response, which was blocked with either L-N(G) -Nitroarginine methyl ester (0.5 m m) or 1 m m of lidocaine. PHY-N (1, 3 and 10 µg ml(-1) ) induced an increase in tonus of the biventer-cervicis neuromuscular preparation that was blocked with pre-treatment of galamine (10 µ m). Administration of 6 mg kg(-1) PHY-N intramuscularly produced death in broilers by spastic paralysis. In conclusion, PHY-N induces nerve depolarization and nonspecifically increases neurotransmitter release.

[Lipid variation in oocytes of the jellyfish Stomolophus meleagris (Scyphozoa: Rhizostomeae) from Las Guasimas Lagoon, Mexico, during gonadal development]. / Variación lipídica en los ovocitos de la medusa Stomolophus meleagris (Scyphozoa: Rhizostomeae), durante el desarrollo gonádico, en la laguna Las Guásimas, Sonora, México.

The jellyfish Stomolophus meleagris has potential for commercial exploitation but there is little information on their reproductive biology. This paper seeks to evaluate some biochemical and demographic characteristics of the species. Samples were taken monthly during 2005 and 2006. Jellyfish collected in 2005 were used to describe the characteristics and quantity of oocyte triglycerides and phospholipids with the Sudan black technique, and to ascertain the degree of gonadal development and sex ratio by the hematoxylin-eosin technique. The 2006 jellyfish were used to determine the size at first maturity and protein and total lipids contents. Four stages of development in both sexes were determined, with a continuous gamete development. The highest percentage of mature organisms was recorded in April. The proportion of sexes was 0.7:1.3. We found higher concentrations of triglycerides than phospholipids in the cytoplasm. There was a positive correlation between triglycerides and the diameter of the oocyte. The size at first maturity for both sexes was 105 mm. The highest protein and lipids contents were obtained in April and March respectively.

Variación lipídica en los ovocitos de la medusa Stomolophus meleagris (Scyphozoa: Rhizostomeae), durante el desarrollo gonádico, en la laguna Las Guásimas, Sonora, México

Lipid variation in oocytes of the jellyfish Stomolophus meleagris (Scyphozoa: Rhizostomeae) from Las Guasimas Lagoon, Mexico, during gonadal development. The jellyfish Stomolophus meleagris has potential for commercial exploitation but there is little information on their reproductive biology. This paper seeks to evaluate some biochemical and demographic characteristics of the species. Samples were taken monthly during 2005 and 2006. Jellyfish collected in 2005 were used to describe the characteristics and quantity of oocyte triglycerides and phospholipids with the Sudan black technique, and to ascertain the degree of gonadal development and sex ratio by the hematoxylin-eosin technique. The 2006 jellyfish were used to determine the size at first maturity and protein and total lipids contents. Four stages of development in both sexes were determined, with a continuous gamete development. The highest percentage of mature organisms was recorded in April. The proportion of sexes was 0.7:1.3. We found higher concentrations of triglycerides than phospholipids in the cytoplasm. There was a positive correlation between triglycerides and the diameter of the oocyte. The size at first maturity for both sexes was 105mm. The highest protein and lipids contents were obtained in April and March respectively. Rev. Biol. Trop. 58 (1): 119-130. Epub 2010 March 01.

La medusa S. meleagris, ha mostrado potencial de explotación pero hay escasa información sobre su biología reproductiva. El presente trabajo pretende conocer el contenido de los triglicéridos y fosfolípidos en los ovocitos durante el desarrollo gonadal, así como la proporción de sexos, talla de primera madurez y la concentración de proteínas y lípidos totales en la medusa. Se realizaron muestreos mensuales durante 2005 y 2006. A las medusas recolectadas en 2005, se les aplicó la técnica del sudán negro, para describir las características del ovocito y la cantidad triglicéridos y fosfolípidos y la técnica de hematoxilina-eosina para conocer el grado de desarrollo gonádico y la proporción de sexos. Las medusas del 2006 se emplearon para determinar la talla de primera madurez y el contenido de proteínas y lípidos totales. Se observaron cuatro fases de desarrollo en ambos sexos, con un desarrollo gamético continúo. El mayor porcentaje de organismos maduros se registró en abril. La proporción de sexos fue de 0.7:1.3. Se encontró mayor concentración de triglicéridos que de fosfolípidos en el citoplasma. Se obtuvo una correlación positiva entre triglicéridos y el diámetro del ovocito. La talla de primera madurez para ambos sexos fue de 105 mm. El mayor contenido de proteínas se obtuvo en abril y para lípidos en marzo.

Glyco- and sphingophosphonolipids from the medusa Phyllorhiza punctata: NMR and ESI-MS/MS fingerprints.

The medusa Phyllorhiza punctata has been found in Brazilian waters where it is an exotic species, having arrived in ballasts from the Indo-Pacific Ocean in the general region of North Australia and Indonesia. Fatty acids of the intact animal and its component umbrella, oral arms, and mucus were identified. Two different groups of glycolipids and a sphingolipid were isolated by silica-gel column chromatography and characterized using GC-MS, ESI-MS, 1D, 2D (13)C, (1)H and (31)P NMR spectroscopy. They were sulfoquinovosyldiacylglycerol (SQDG), monogalactosyldiacylglycerol (MGDG), and ceramide aminoethylphosphonate (CAEP). The CAEP long chain base (LCB) and its polar head group (PHG) formed by partial hydrolysis, were analyzed by ESI-MS/MS. The probable origin of MGDG and SQDG in the jellyfish is the result of an endosymbiotic association with a microalga of the Dinoflagellate group, since these lipids are commonly found in photosynthetic membranes.

Toxicity and mAChRs binding activity of Cassiopea xamachana venom from Puerto Rican coasts.

A separation of toxic components from the upside down jellyfish Cassiopea xamachana (Cx) was carried out to study their cytotoxic effects and examine whether these effects are combined with a binding activity to cell membrane receptors. Nematocysts containing toxins were isolated from the autolysed tentacles, ruptured by sonication, and the crude venom (CxTX) was separated from the pellets by ultracentrifugation. For identifying its bioactive components, CxTX was fractionated by gel filtration chromatography into six fractions (named fraction I-VI). The toxicity of CxTX and fractions was tested on mice; however, the hemolytic activity was tested on saline washed human erythrocytes. The LD50 of CxTX was 0.75 microg/g of mouse body and for fraction III, IV and VI were 0.28, 0.25 and 0.12 microg/g, respectively. Fractions I, II and V were not lethal at doses equivalent to LD50 1 microg/g. The hemolytic and phospholipase A2 (PLA2) activities of most fractions were well correlated with their mice toxicity. However, fraction VI, which contains the low molecular mass protein components (< or =10 kDa), has shown no PLA2 activity but highest toxicity to mice, highest hemolytic activity, and bound significantly to the acetylcholine muscarinic receptors (mAChRs) isolated from rat brain. The results suggested that fraction VI contains proteinaceous components contributing to most of cytolysis as well as membrane binding events. Meanwhile, fraction IV has shown high PLA2 that may contribute to the venom lethality and paralytic effects.