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ABSTRACT: Observations of gonads and oocyte development stages (OS) have been achieved in Calanus helgolandicus females fed different algal diets and starved in filtered sea water under laboratory conditions during 8days. The effects
of 20 diets on egg production rates (EPR), hatching success (HS) and proportion of abnormal larvae (AL, development stages
N1-2) were examined. With the control diet Prorocentrum minimum EPR and HS values were high, while AL was very low, coinciding with intact cell structures in oogonia (OO) and normal OS
(OS1-OS4). With the other diets, oocyte maturation, EPR, HS and AL patterns were partially or totally impaired. Decrease of
EPR coincided with the arrest of OS3 maturation and oocyte degradations, characterised by cell fragmentation, presence of
apoptotic bodies in the OS3, degradation of cytoplasm in OS3 and OS4 and consequently the arrest of OS4 formation. These degradations
were reversible when females were fed a favourable diet. Results reflect the presence of two distinct inhibitory mechanisms.
Inhibition mechanism (1) impairs egg production. It was induced by starvation or by several species belonging to Bacillariophyceae
(Chaetoceros calcitrans, Guinardia delicatula, Guinardia
striata, Odontella regia, Rhizosolenia setigera, Stephanopyxis turris, Thalassiosira pseudonana) and mixed-diatom assemblages (collected in the field) and to the prymnesiophyte Pavlova lutherii. Remarkably other diatoms like Navicula sp., Nitzschia sp., Skeletonema costatum and Thalassiosira rotula did not induce mechanism (1) Inhibitory mechanism (2) affected exclusively HS and AL and was triggered by species independent
of the production of polyunsaturated aldehydes (PUA), which are supposed to have adverse impacts on HS and larval development.
Marine Biology 04/2012; 152(2):415-428. · 2.28 Impact Factor
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ABSTRACT: DNA integrity checkpoint control was studied in the sea urchin early embryo. Treatment of the embryos with genotoxic agents such as methyl methanesulfonate (MMS) or bleomycin induced the activation of a cell cycle checkpoint as evidenced by the occurrence of a delay or an arrest in the division of the embryos and an inhibition of CDK1/cyclin B activating dephosphorylation. The genotoxic treatment was shown to induce DNA damage that depended on the genotoxic concentration and was correlated with the observed cell cycle delay. At low genotoxic concentrations, embryos were able to repair the DNA damage and recover from checkpoint arrest, whereas at high doses they underwent morphological and biochemical changes characteristic of apoptosis. Finally, extracts prepared from embryos were found to be capable of supporting DNA repair in vitro upon incubation with oligonucleotides mimicking damage. Taken together, our results demonstrate that sea urchin early embryos contain fully functional and activatable DNA damage checkpoints. Sea urchin embryos are discussed as a promising model to study the signaling pathways of cell cycle checkpoint, DNA repair and apoptosis, which upon deregulation play a significant role in the origin of cancer.
Cellular and Molecular Life Sciences CMLS 08/2007; 64(13):1723-34. · 6.57 Impact Factor
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ABSTRACT: Two double-labelling methods, Tunel+propidium iodide and AnnexinV-FITC+propidium iodide, have been tested to diagnose cell degradation processes in N1–N2 nauplius stages produced by spawning females of the copepod Calanus helgolandicus fed either non-toxic (PRO: control) or toxic (TR1) diets under laboratory conditions. Observation of labelled samples with a confocal laser-scanning microscope revealed that the maternal-food effect, following absorption of the toxic diet, induced cell apoptosis and necrosis in 80–100% of the offspring, which sooner or later died. Similar cell damages were also observed in 18–50% of the nauplii produced by females incubated in filtered seawater. Such cell degradation processes and high mortality were not observed in nauplii produced by females fed the non-toxic diet. Our protocols allowed detection of cell degradations before death of unhealthy larvae, preferentially with a confocal microscope or a conventional visible-fluorescent-light microscope.
Marine Biology 01/2003; 143(5):889-895. · 2.28 Impact Factor
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ABSTRACT: Expression of heat shock proteins (hsp) is a homeostatic mechanism induced in both prokaryotic and eukaryotic cells in response to metabolic and environmental insults. A growing body of evidence suggests that in mammals, the hsp response is integrated with physiological responses through neuroendocrine signaling. In the present study, we have examined the effect of noradrenaline (NA) on the hsp70 response in mollusc immune cells. Oyster and abalone hemocytes transfected with a gene construct containing a gastropod hsp70 gene promoter linked to the luciferase reporter-gene were exposed to physiological concentrations of NA, or to various alpha- and beta-adrenoceptor agonists and antagonists. Results show that NA and alpha-adrenergic stimulations induced the expression of luciferase in transfected mollusc immunocytes. Furthermore, exposure of hemocytes to NA or to the alpha-adrenoceptor agonist phenylephrine (PE) resulted in the expression of the inducible isoform of the hsp70 protein. Pertussis toxin (PTX), the phospholipase C (PLC) inhibitor U73122, the protein kinase C (PKC) inhibitor calphostin C, the Ca(2+)-dependent PKC inhibitor Gö 6976 and the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor LY294002 blocked the PE-mediated induction of the hsp70 gene promoter. These results suggest that alpha-adrenergic signaling induces the transcriptionnal upregulation of hsp70 in mollusc hemocytes through a PTX-sensitive G-protein, PLC, Ca(2+)-dependent PKC and PI 3-kinase. Thus, a functional link exists between neuroendocrine signaling and the hsp70 response in mollusc immune cells.
Journal of Cell Science 11/2001; 114(Pt 19):3557-64. · 6.11 Impact Factor
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ABSTRACT: Juvenile oysters Crassostrea gigas cultured in the Bay of Morlaix (France) have suffered unexplained summer mortalities for over a decade. In the present study, we tested the hypothesis that a bacterial pathogen could be responsible for this phenomenon. A first attempt failed to isolate a bacterial pathogen from moribund or weak oysters. Only non-pathogenic, probably opportunistic, bacteria were isolated. As an alternative approach, we focused on oysters presenting reduced stress-response capacities (determined by circulating noradrenaline measurements), a characteristic of juvenile oysters entering an early phase of the disease. Cultures of bacterial isolates on TCBS plates revealed that a Vibrio strain was present in diseased oysters and scarce or absent in healthy oysters. Experimental infections indicated that this Vibrio can cause mortalities of juvenile oysters when injected at concentrations ranging from 10(4) to 10(8) CFU oyster(-1). Similarly to the summer mortality disease, the Vibrio isolate caused higher mortalities at higher temperatures; apparently, it could not be transmitted horizontally, it did not affect adult oysters and it induced stress-response dysfunctions in juvenile oysters. Phenotypic and genotypic characterizations identified the pathogen as Vibrio splendidus. Taken together, the present results satisfy Koch's postulate and suggest that this bacterial strain is probably responsible for the juvenile oyster summer mortalities in the Bay of Morlaix.
Diseases of Aquatic Organisms 10/2001; 46(2):139-45. · 2.20 Impact Factor
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ABSTRACT: Catecholamines (CA) regulate several physiological processes in molluscs. Experiments have been conducted to determine the effects of noradrenaline (NA), the principal CA circulating in bivalve hemolymph, on oyster hemocytephagocytosis. Results show that NA had a dose-dependent inhibitory effect on phagocytosis at physiological concentrations of 0.1 microM and above. The beta-adrenoceptor agonist isoproterenol mimicked the inhibitory effects of NA on phagocytosis, whereas the alpha-adrenoceptor agonist phenylephrine had no significant effect. Furthermore, the beta-adrenoceptor antagonist propanolol, but not the alpha-adrenoceptor antagonist prazosin, prevented the inhibition of phagocytosis by NA. The type IV phosphodiesterase inhibitor rolipram acted synergistically with a suboptimal concentration of isoproterenol to inhibit phagocytosis, and the protein kinase A inhibitor H-89, but not the protein kinase C inhibitor calphostin C, attenuated the effect of isoproterenol. These results show that NA can modulate oyster hemocyte phagocytosis via a beta-adrenergic receptor/cAMP/protein kinase A signaling pathway.
General and Comparative Endocrinology 07/2001; 122(3):252-9. · 3.27 Impact Factor
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ABSTRACT: Oysters are permanently exposed to various microbes, and their defense system is continuously solicited to prevent accumulation of invading and pathogenic organisms. Therefore, impairment of the animal's defense system usually results in mass mortalities in cultured oyster stocks or increased bacterial loads in food products intended for human consumption. In the present study, experiments were conducted to examine the effects of stress on the juvenile oyster's resistance to the oyster pathogen Vibrio splendidus. Oysters (Crassostrea gigas) were challenged with a low dose of a pathogenic V. splendidus strain and subjected to a mechanical stress 3 days later. Both mortality and V. splendidus loads increased in stressed oysters, whereas they remained low in unstressed animals. Injection of noradrenaline or adrenocorticotropic hormone, two key components of the oyster neuroendocrine stress response system, also caused higher mortality and increased accumulation of V. splendidus in challenged oysters. These results suggest that the physiological changes imposed by stress, or stress hormones, influenced host-pathogen interactions in oysters and increased juvenile C. gigas vulnerability to Vibrio splendidus.
Applied and Environmental Microbiology 06/2001; 67(5):2304-9. · 3.83 Impact Factor
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ABSTRACT: Catecholamines (CA) are known to be present in the microenvironment of molluscan immunocytes. In the present study, experiments were conducted to determine the effects of noradrenaline (NA), the principal CA circulating in bivalve hemolymph, on the luminol-dependent chemiluminescence (CL) of oyster Crassostrea gigas hemocytes. Results show that NA had a dose-dependent inhibitory effect on the CL-response at the physiological concentration of 0.1 microM and above. The alpha-adrenoceptor agonist phenylephrine had no significant effect on the CL-response whereas the beta-adrenoceptor agonist isoproterenol mimicked the inhibitory effects of NA on the CL-response. The beta-adrenoceptor antagonist propanolol, but not the alpha-adrenoceptor antagonist prazosin, prevented the negative effects of NA on the CL-response. Taken together, these results show that beta-adrenergic receptors are present at the surface of oyster hemocytes and allow NA to down-regulate the CL-response.
Developmental & Comparative Immunology 06/2001; 25(4):285-9. · 3.27 Impact Factor
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ABSTRACT: Failure of female reproductive capacity in the copepod Calanus helgolandicus was related to number and combination of the phytoplankton species in the diets. The maternal food effects were detectable at different levels: fecundity, oogenesis and hatching. Fecundity and hatching were normal with two single (ca. Isochrysis galbana and Prorocentrum minimum) and one mixed (Phaeodactylum tricornutum+Dunaliella tertiolecta+Pavlova lutherii+I. galbana+P. minimum) diets. With the single P. lutherii diet, fecundity decreased, but hatching remained optimal. The daily egg production and hatching rates decreased significantly in females fed the other single P. tricornutum, D. tertiolecta and mixed (P. tricornutum+D. tertiolecta+P. lutherii+I. galbana) diets, or starved. The fecundity decrease coincided with gonad atresia, which was reversible when P. tricornutum and P. lutherii diets were replaced by P. minimum diet. It was irreversible when D. tertiolecta was replaced by P. minimum, leading to female sterilization expressed by the deterioration of OS3 and OS2 oocytes, as a function of the feeding duration. We assume that atresia of female gonads was caused by the limitation of essential nutrients in food, such as fatty acids, which induced catabolism and recycling of yolk reserves and thus, maintenance of gonad integrity and low spawning rates. With the D. tertiolecta diet, abnormally high increase of ornithine concentrations in eggs showed that the ornithine metabolism and polyamine pathway were affected during oogenesis, leading atresia of oocytes to be deeply disturbed and followed up by necrosis of the gonads.
Journal of Experimental Marine Biology and Ecology 05/2001; 259(1):85-107. · 1.88 Impact Factor
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ABSTRACT: The stress response is a series of coordinated physiological reactions increasing an organism's capacity to maintain homeostasis in the presence of threatening agents. This fundamental process is known to involve hormonal signaling to rapidly modulate key physiological functions in vertebrates, but data are lacking concerning neuroendocrine responses to stress in invertebrates. The present study examined circulating catecholamine (CA) responses to stress in oysters. Mechanical disturbances (consisting of shaking the animals) and temperature or salinity variations were applied to the animals because these three types of stressors are commonly encountered by oysters in aquaculture or in their natural habitat. Results show that both circulating noradrenaline (NA) and dopamine (DA) concentrations increased in response to stress. The catecholaminergic response to acute mechanical stressors was rapid (less than 5 min), transient (a return to basal CA levels was observed after 60-90 min), and reflected both the intensity and duration of the perturbation. In contrast, responses to temperature and salinity variations were long lasting (up to 72 h). CA concentrations varied from 1.61 +/- 0.30 ng NA/ml and 0.41 +/- 0.05 ng DA/ml to maximal values of 22.07 +/- 0.97 ng NA/ml and 2.24 +/- 0.19 ng DA/ml. Such CA concentrations are known to induce physiological responses in bivalves, suggesting that stress-induced NA and DA changes exert a regulatory function in oysters.
General and Comparative Endocrinology 05/2001; 122(2):181-8. · 3.27 Impact Factor
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ABSTRACT: Catecholamines and pro-opiomelanocortin (POMC)-derived peptides, some of the central regulators of the stress-response systems of vertebrates, are also present in invertebrates. However, studies are needed to determine how these hormones participate in the organisation of neuroendocrine stress-response axes in invertebrates. Our present work provides evidence for the presence of an adrenergic stress-response system in the oyster Crassostrea gigas. Noradrenaline and dopamine are released into the circulation in response to stress. Storage and release of these hormones take place in neurosecretory cells presenting morphological and biochemical similarities with vertebrate chromaffin cells. Both in vivo and in vitro experiments showed that applications of the neurotransmitters acetylcholine or carbachol caused no significant release of noradrenaline or dopamine. Moreover, the nicotinic antagonists hexamethonium and &agr; -bungarotoxin and the muscarinic antagonist atropine caused no significant inhibition of catecholamine release in stressed oysters. Adrenocorticotropic hormone (ACTH) induced a significant release of noradrenaline, but the release of dopamine in response to ACTH was not significant. These results suggest that, unlike that of vertebrates, the adrenergic stress-response system of oysters is not under the control of acetylcholine and that other factors, such as the neuropeptide ACTH, might control this system.
Journal of Experimental Biology 05/2001; 204(Pt 7):1247-55. · 3.00 Impact Factor
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ABSTRACT: The contents of free amino acid (FAA) and total amino acid (TAA) pools were determined in the eggs, embryos and N1 and N2
nauplii generated by Calanus helgolandicus females fed either the diatom Phaeodactylum tricornutum or the dinoflagellate Prorocentrum minimum for 3 and 10 d. With both diets, egg production rates increased by a factor of 5 to 10, and free amino acid (FAA) and total
amino acid (TAA) contents were double those measured in eggs spawned by wild females. Higher levels were measured for almost
all amino acids, except methionine, taurine, glutamine, glutamic acid, aspartic acid and ornithine. When embryos developed
to live nauplii, 50 to 72% of the initial concentration of the FAA content was used. There was no preferential utilisation
of essential over non-essential amino acids. The TAA pool also varied with development. The mean FAA:TAA ratio remained between
11 and 19. With the Prorocentrum minimum diet, hatching success remained constantly >85%, whereas it declined to 0% at the end of the 10 d incubation period with
Phaeodactylum tricornutum. After 10 d of feeding on this alga, the FAA content of developing embryos increased significantly, indicating liberation
of amino acids due to proteolysis. None of these embryos developed to hatching. Inhibition of hatching success related to
the ingestion of P. tricornutum was not due to a lack of any amino acid. The results indicate that the chemical composition of freshly spawned copepod eggs
is sensitive to the maternal diet.
Marine Biology 05/1999; 134(1):89-98. · 2.28 Impact Factor
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ABSTRACT: ABSTRACT: Egg production rates (EPR) by Calanus helgolandicus females were investigated with specimens sampled weekly, from April to November 2003 and from March to October 2004, at a station located in the English Channel off Roscoff. Comparison of results between 1994, 2003 and 2004 showed that C. helgolandicus was a late spawner in 1994 and became an early spawner in 2003 and 2004. In all cases high variations in EPR were observed, which could not be correlated to phytoplankton biomass, expressed as diatom, chlorophyll a, particulate carbon and nitrogen concentrations in 2003 and 2004. Neither were they correlated to food quality, expressed as C/N ratio. To explain this mismatch between EPR and food concentration, a series of mixed phytoplankton species dominated by diatoms (≥11 μm filtrate representing natural diatom assemblages: NDA) and 7 single diatom species, all occurring during blooms in the field, were assayed as diets with C. helgolandicus females. Ingestion of diatoms by females was estimated by faecal pellet production rates and complementary scanning electron microscopy examinations of diatom remains in pellets. Depending on diatom species in diets, EPR was either increased or depressed 2 to 3 d after food uptake by females had started. The EPR decrease was reversible, when diatom diets were replaced by the dinoflagellate Prorocentrum minimum. This effect was also observed when females were transferred to natural phytoplankton populations from the English coast of the Channel close to Plymouth, where food composition in the field differed compared to that off Roscoff. EPR ceased completely when the concentration of NDA diets was artificially increased, but recovered after a shift to a dinoflagellate diet. These results indicate that phytoplankton dominated by diatoms can impair C. helgolandicus egg production in the field. This effect was not related to the production of polyunsaturated aldehydes by diatoms. Limitations due to unidentified essential compounds not provided by the metabolism of diatoms, or unknown diatom-derived toxins, were probably involved.
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ABSTRACT: Field estimates of the production and viability of eggs of the copepod Calanus helgolandicus were conducted weekly during 1993 and 1994 at two fixed stations in the western English Channel. The stations were located in the coastal zone off Roscoff (NW France) and Plymouth (SW England). These areas are characterized by homogeneous and seasonally stratified waters, respectively. In these two different environments, the egg production rate varied seasonally and was significantly correlated with chlorophyll a concentration, but not with surface temperature. Significant correlations between fecundity and particulate carbon and nitrogen were found only off Plymouth. The seasonal pattern of fecundity differed between the two sites. Off Plymouth, spawning occurred earlier, lasted longer and was significantly higher than in Roscoff waters. In both localities, fecundity was generally below the maximum specific egg production rate (∼60 eggs female−1 day−1 implying that adult females were permanently under, or close to, limiting food conditions. Egg viability showed seasonal variations in both localities, ranging between 20 and 100%. However, in 1994, the amplitude of egg viability variation was lower in stratified waters. The results show that the potential fecundity of C.helgolandicus was different between mixed and stratified waters of the western English Channel. Food quality as well as quantity appear to be important for the reproductive success of this copepod species.
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Marine Ecology - Progress Series, v.245, 33-45 (2002).
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ABSTRACT: Catecholamines (CA) are known to be present in the microenvironment of molluscan immunocytes. In the present study, experiments were conducted to determine the effects of noradrenaline (NA), the principal CA circulating in bivalve hemolymph, on the luminol-dependent chemiluminescence (CL) of oyster Crassostrea gigas hemocytes. Results show that NA had a dose-dependent inhibitory effect on the CL-response at the physiological concentration of 0.1 μM and above. The α-adrenoceptor agonist phenylephrine had no significant effect on the CL-response whereas the β-adrenoceptor agonist isoproterenol mimicked the inhibitory effects of NA on the CL-response. The β-adrenoceptor antagonist propanolol, but not the α-adrenoceptor antagonist prazosin, prevented the negative effects of NA on the CL-response. Taken together, these results show that β-adrenergic receptors are present at the surface of oyster hemocytes and allow NA to down-regulate the CL-response.
Developmental & Comparative Immunology.
