Geographical patterns of dominant bivalves and a polychaete in Europe: no metapopulations in the marine coastal zone?

The genetic diversity, differentiation and performance of some dominant invertebrates in the marine coastal zone of Europe are reviewed in order to discuss the use of the metapopulation concept in the marine coastal realm. A consistently high genetic diversity of the species studied (mussels of the Mytilus edulis complex, Baltic clams Macoma balthica and lugworms Arenicola marina), a low differentiation and an almost uniform ecophysiological performance (determined by growth, maximum length, level reserve constituents or stress resistance) all along the coast of Europe do not support the use of the metapopulation concept. Electronic Publication     

Comparison of the two different phosphagen systems in the lugworm Arenicola marina

The different phosphagen systems in the lugworm Arenicola marina, the phosphotaurocyamine/taurocyamine kinase system of the body wall and the phosphocreatine/creatine kinase system of the spermatozoa, have been investigated to answer the question whether the change reflects different functional modes of these phosphagen systems. Enzyme analyses have shown that in contrast to the body wall taurocyamine kinase, creatine kinase of spermatozoa exists in at least two different forms which are compartmented in the mitochondria (creatine kinase I) and in the flagellum (creatine kinase II). Creatine kinase I is strongly attached to cell structures which require detergents and high phosphate concentrations for solubilization. The affinities of taurocyamine kinase and creatine kinase for all substrates are very similar except the extremely high K _m for creatine of both creatine kinase I and II. The level of creatine in spermatozoa is fivefold higher than taurocyamine in the body wall at similar phosphorylation potential (ATP/ADO_free) and ATP-buffer capacity (phosphagen/ATP), reflecting the higher equilibrium constants of the creatine kinase reaction compared to that of the taurocyamine kinase reaction (Ellington 1989). The high creatine concentration gives the phosphocreatine/creatine kinase system an advantage over the phosphotaurocyamine/taurocyamine kinase system for transport of energyrich phosphate at high phosphorylation potential by increasing the radial diffusion flux. The maximum diffusive flux of free ADP in spermatozoa is three orders of magnitude below the respiratory ATP production while the creatine flux would allow an unlimited energy transport over the long diffusion distance. In lugworm body wall, however, the low ATP turnover and the low diffusion distances between mitochondria and myosin-ATPases do not require a phosphagen shuttle.Abbreviations ADP _free cytoplasmic adenosine diphosphate - Ap ₅ A P₁, P₅-di(adenosine-5-) pentaphosphate - AK arginine kinase - CK creatine kinase (EC 2.7.3.2) - DTT dithiothreitol - GAPDH glyceraldehydephosphate dehydrogenase (EC 1.2.1.12) - HOADH 3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) - IEP isoelectric point - MIM mitochondria isolating medium - P _i-free cytoplasmic inorganic phosphate - (P)Arg (phospho)arginine - (P)Cr (phospho)creatine - (P)Tc (phospho)taurocyamine - SEM scanning electron microscopy - TK taurocyamine kinase - TEM transmission electron microscopy     

Glycan-binding profile of a D-galactose binding lectin purified from the annelid, Perinereis nuntia ver. vallata

A lectin recognizing D-galactose was purified from the pacific annelid Perinereis nuntia ver. vallata (Polychaeta) by affinity chromatography. Hemagglutinating activity, with a very low titer suggesting the presence of lectin appeared in the supernatant from the homogenization of body with Tris-buffered saline. However, dialyzed supernatant from the precipitate homogenized by galactose in the buffer revealed strong hemagglutinating activity against human erythrocytes. The crude supernatant was applied onto lactosyl–agarose column, and only the supernatant eluted from precipitate with galactose was obtained a galactose-binding lectin with 32 kDa polypeptide was obtained from the supernatant of the precipitate, extracted in presence of galactose. It suggests that the lectin tightly binds with glycoconjugate as endogenous ligand(s) in the tissue. Hemagglutinating activity against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-galactose, N-acetyl-D-galactosamine, lactose, melibiose, and asialofetuin. Glycan-binding profile of the lectin analyzed by frontal affinity chromatography shows that the lectin recognizes branched complex type N-linked oligosaccharides and both type 1 (Galβ1-3GlcNAc) and type 2 (Galβ1-4GlcNAc) lactosamine. The surface plasmon resonance study of the lectin against asialofetuin showed the k_ass and k_diss values are 5.14 × 10⁴ M^− 1 s^− 1 and 2.9 × 10⁻³ s^− 1, respectively. The partial primary structure of the lectin reveals 182 amino acids with novel sequence.     

Recombinant expression, synthesis, purification, and solution structure of arenicin

Arenicins are 21-residue cationic antimicrobial peptides, isolated from marine polychaeta Arenicola marina. In order to determine a high-resolution three-dimensional structure of arenicin-2, the recombinant peptide was overexpressed as a fused form in Escherichia coli. Both arenicin isoforms were synthesized using the Fmoc-based solid-phase strategy. Recombinant and synthetic arenicins were purified, and their antimicrobial and spectroscopic properties were analyzed. NMR investigation shows that in water solution arenicin-2 displays a prolonged beta-hairpin, formed by two antiparallel beta-strands and stabilized by one disulfide and nine hydrogen bonds. A significant right-handed twist in the beta-sheet is deprived the peptide surface of amphipathicity. CD spectroscopic analysis indicates that arenicin-2 binds to the SDS and DPC micelles, and conformation of the peptide is significantly changed upon binding. Arenicin strongly binds to anionic lipid (POPE/POPG) vesicles in contrast with zwitterionic (POPC) ones. These results suggest that arenicins are membrane active peptides and point to possible mechanism of their selectivity toward bacterial cells.     

Extension of the breeding season and its effects on fertilization and development in two species of lugworm (Arenicola marina and A. defodiens)

In the autumn/winter breeding polychaetes, Arenicola marina and A. defodiens, spawning can be advanced or delayed by a number of months through temperature manipulation of the adults. However, this manipulation may have significant consequences for fertilization rates and embryo developmental success and so in vitro fertilizations were performed to assess the impact of manipulation. Firstly, we used oocytes and sperm obtained from advanced or delayed individuals. For both species, using gametes from 4 weeks advanced individuals did not result in a significant reduction in development, however, gametes from individuals advanced (A. marina only) or delayed by 8 weeks resulted in significantly fewer embryos developing normally. Reciprocal crosses of temperature-manipulated A. marina gametes (from 4 weeks advanced and 4 weeks delayed individuals) with those at the natural spawning time confirmed that the reduction in developmental success in both was attributable to the oocytes. After 5 h post-fertilization, the majority of oocytes from advanced individuals had fertilized, but by 24 h most were abnormal. For fertilizations with gametes from delayed individuals, nearly 100% of the embryos were developing normally after 24 h, but after 144 h significantly more were abnormal in crosses involving oocytes from delayed females. Although both species have reproductive plasticity to extend their breeding season, the significant reduction in the numbers of competent larvae produced as the spawning is delayed or advanced further may be a significant bottleneck in aquaculture and it may also have considerable implications for the long-term reproductive success of a population in response to environmental change.Sympatric populations of the species exist in many locations and the inherent variability in the breeding seasons could allow spawning times to overlap. Artificially delaying A. marina individuals enabled fertilizations to be performed with A. defodiens at the natural spawning time in the laboratory. Both conspecific fertilizations produced 100% trochophore larvae after 120 h, but A. defodiens oocytes failed to fertilize after incubation with A. marina sperm, in comparison to the A. marina oocytes incubated with A. defodiens sperm where 40% developed to the trochophore stage. This asymmetric gamete incompatibility may be one of a suite of mechanisms to minimise hybridisation.     

In vivo nuclear magnetic resonance studies on the lugworm Arenicola marina. I. Free inorganic phosphate and free adenylmonophosphate concentrations in the body wall and their dependence on hypoxia

The cytoplasmic concentrations of free inorganic phosphate and free AMP in the body wall of the lugworm Arenicola marina were estimated in order to verify their proposed regulatory role in glycogenolysis during anaerobiosis (Kamp 1993). Using in vivo ³¹P nuclear magnetic resonance spectroscopy the concentration of free inorganic phosphate was determined to be 4.7±0.7 mmol·1^-1 (±standard deviation, n=3) varying with season (Juretschke and Kamp 1995). These values were two to three times lower than those measured in perchloric acid extracts. In contrast, values for the phosphagen phosphotaurocyamine assessed biochemically in the extracts and by in vivo nuclear magnetic resonance were very similar. During the transition from normoxia to hypoxia the concentration of free inorganic phosphate increased to the same extent and at the same rate as the concentration of phosphotaurocyamine decreased. A discrepancy was also found for the biochemically determined AMP and ADP concentrations in the extract and those derived from the equilibrium constants of the taurocyamine kinase (ADP_free) and adenylate kinase (AMP_free) reactions. Calculated concentrations of ADP_free and AMP_free in normoxic specimens were about two or even four orders of magnitude lower than the values determined in extracts. During hypoxia the concentrations of AMP and ADP increase moderately when measured biochemically in extracts, while the values for ADP_free and AMP_free rise three- and nine-fold during the first 3 h of hypoxia. Thereafter, the levels stay constant due to a progressive acidosis. If during hypoxia pH_i is stabilized by addition of buffering substances to the incubation medium, both ADP_free and AMP_free rise continuously. The significant changes found for the concentrations of free inorganic phosphate and AMP_free support their assumed regulatory role in glycogenolysis during anaerobiosis, though these AMP_free values seem too low to actually activate glycogen phosphorylase. The strong effect of pH_i on the levels of ADP_free and AMP_free suggest a mechanism by which acidosis prevents a continuing increase of glycogenolysis (ATP-producing pathway) during prolonged anaerobiosis (protective effect of acidosis).Abbreviations ADP _free cytoplasmic adenosine diphosphate - AK adenylate kinase - AMP _free cytoplasmic adenosine monophosphate - CK creatine kinase - GPase glycogen phosphorylase - MW molecular weight - Int P _i integral of P_i-signal - NMR nuclear magnetic resonance - P _i-free cytoplasmic inorganic phosphate - PCA perchlori acid - PFK 6-phosphofructokinase - PME phosphomonoester - PPA phenylphosphonic acid - (P)Tc (phospho)taurocyamine - S _f saturation factor - Sw sea water - TcK taurocyamine kinase - TRA triethanolamine - TRIS tris(hydroxymethyl)aminomethane     

In vivo nuclear magnetic resonance studies on the lugworm Arenicola marina. II Seasonal changes of metabolism

In vivo ³¹P- and ¹³C-NMR spectra of the lugworm Arenicola marina (Polychaeta, Annelida) gathered between 1988 and 1994 at different times of the year were evaluated for seasonally dependent metabolic changes. Beside the typical ³¹P-NMR signals of ATP and (phospho)taurocyamine, other seasonally dependent signals were observed: from January to March an intensive signal at 1.4–1.8 ppm was identified as inorganic phosphate compartmented in an acidic intestinal lumen. Between April and September signals at 1.2–1.4 ppm were assigned to phosphodiester. Starting in July males showed a second phosphagen signal [(phospho)creatine of spermatozoa, cf. Kamp and Juretschke (1989a)] whose intensity increased until spawning in September. The (phospho)taurocyamine/ATP ratio was also dependent on the season. In January or February the ratio reached 11, while in summer and autumn the ratio was between 4 and 5. As verified by biochemical assays this effect was caused by a significant decrease of ATP in the lugworm body wall during winter (December–February). The phosphagen (phospho)taurocyamine and the respective unphosphorylated guanidine taurocyamine remained constant throughout the year. Levels of free inorganic phosphate incurred similar changes to ATP. ¹³C-NMR spectra of lugworms showed a dramatic change in lipid stores. They were below the detection limit between January and March but developed into the most intensive signals during summer. The most abundant amino acids, glycine and alanine, were observed throughout the year while glycogen could not be detected in the ¹³C-NMR spectra. After treating tissue extracts with amyloglucosidase, the signals of the hydrolytic product glucose were recorded indicative of NMR-invisible glycogen stores.Abbreviations AN adenine nucleotides - NMR nuclear magnetic resonance - P _i inorganic phosphate - (P)Cr (phospho)creatine - PDE phosphodiester - PME phosphomonoester - PPA phenylphosphonic acid - (P)Tc (phospho)taurocyamine     

Contrasting breeding periodicity of nearby populations of the lugworm,Arenicola marina (Annelida,Polychaeta)

Summary

The breeding cycles of lugworm taken from three locations on the east coast of Ireland have been investigated through examination of coelomic fluid samples which contain the gametes, taken at monthly intervals. These reveal major differences in the timing and duration of breeding between the populations despite their geographic propinquity. The differences in breeding cycle cannot be ascribed to varietal or species differences and may arise from the different habitats occupied by the lugworm populations studied.