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At least 19 recordsLinked to original sources
PubMed2004

Nitrite toxicity to crayfish, Astacus leptodactylus, the effects of sublethal nitrite exposure on hemolymph nitrite, total hemocyte counts, and hemolymph glucose.

The 48-h acute toxicity range of nitrite to narrow-clawed crayfish, Astacus leptodactylus was within 22 and 70 mg L(-1) (mean 29.43 mg L(-1)). Environmental chloride (100 mg L(-1) chloride) increased the 48-h toxicity of nitrite to a range of 31 and 80 mg L(-1) (mean 49.20 mg L(-1)). Hemolymph nitrite, total hemocyte counts (THCs), and hemolymph glucose were examined in A. leptodactylus exposed to different sublethal nitrite concentrations. The same parameters were also determined for A. leptodactylus exposed to different sublethal nitrite concentrations with additional environmental chloride. Additionally, hemolymph nitrite and THCs were analyzed for crayfish exposed to nitrite-free water after 24 h following a 48-h exposure to nitrite. In the nitrite-exposed tests, hemolymph nitrite increased directly with water nitrite; however, after recovery, nitrite in hemolymph decreased. In the nitrite plus chloride-exposed tests, the accumulation of nitrite in hemolymph was relatively low compared to the nitrite-exposed tests. Thus, hemolymph to environment ratios of nitrite in the nitrite-exposed tests were higher than those of nitrite plus chloride-exposed tests. THCs decreased following nitrite exposure and, in general, increased after recovery. In the nitrite with chloride exposed and recovery from nitrite tests, THCs increased. Hemolymph glucose levels elevated following nitrite exposure, independent of water nitrite concentrations. However, with environmental chloride nitrite exposure did not cause elevation of hemolymph glucose. Hemolymph nitrite accumulation was found to be closely related to the decrease in THCs and increase in hemolymph glucose.

Animals↗
PubMed2005

Hemolymph clotting in crustaceans: Implications for neuropeptide extraction from invertebrate hemolymph.

The hemolymph of invertebrates contains factors that facilitate clotting as a defense mechanism for injury. However, the clotting process may impair the measurement of hormone titers by interfering with the extraction of peptides. Using hemolymph from freshwater crayfish, our results demonstrate that hemolymph clotting appears to reduce both the amount of an endogenous peptide(s) (Dippu-AST 11-like) extracted from hemolymph, as well as the amount of spiked peptide tracer ([125I]-Dippu-AST 11) recovered from hemolymph. The efficacy of peptide extraction from hemolymph was improved by collecting the hemolymph into a variety of different media prior to hemolymph coagulation. Hemolymph samples collected into media containing the anticoagulant, citrate, had the highest amount of endogenous Dippu-AST 11-like peptide extracted as well as the highest percent recovery of spiked tracer.

Animals↗
PubMed1956

The chemistry of insect hemolymph; organic components of the hemolymph of the silkworm, Bombyx mori, and two other species.

1. Hemolymph was collected for analysis from the silkworm, Bombyx mori, in a series of developmental stages ranging from the second molt to the late pupa. The mean pH of larval hemolymph after collection was found to be 6.45, that of pupal hemolymph, 6.57; in vivo values may be slightly lower. Total dry solids ranged from 5.4 to 10.6 per cent. Total protein ranged from 1.2 to 5.3 per cent, increasing rapidly during the fifth instar. 2. Free amino acids were separated chromatographically and estimated. Of 19 amino acids identified, amounting collectively to 823 to 1497 mg. per 100 ml., glutamine, histidine, and lysine generally occurred in greatest amount. Tryptophan was not detected, and cystine (or cysteine) was found in only one sample. The total free amino acids account for 35 to 55 per cent of the non-protein nitrogen of the plasma. 3. Free sugars, estimated semiquantitatively on chromatograms, comprise glucose, fructose, and sucrose in total amount ranging from about 5 to 40 mg. per 100 ml. Total acid-soluble, ultrafiltrable carbohydrate, estimated as glucose by the anthrone reaction, ranged from 166 to 635 mg. per 100 ml., indicating the presence of low molecular weight sugar derivatives. 4. Inorganic phosphate amounted to 5 to 15 mg. per 100 ml., and acid-soluble organic phosphate to 100 to 200 mg. per 100 ml. The latter fraction includes several substances, of which one was tentatively identified as glucose-6-phosphate and the remainder are as yet unidentified. 5. Single samples of hemolymph were also taken from larvae of the wax moth, Galleria mellonella, and the spruce sawfly, Diprion hercyniae. These contained even higher concentrations of solutes than the silkworm samples, but with a generally similar distribution. The proportions of the free amino acids were different in each species.

Amino Acids↗
PubMed1983

Agglutinins in the horseshoe crab hemolymph: purification of a potent agglutinin of horse erythrocytes from the hemolymph of Tachypleus tridentatus, the Japanese horseshoe crab.

Agglutinins from Tachypleus (Tachypleus tridentatus, the Japanese horseshoe crab) hemolymph were isolated by affinity chromatography on BSM-coupled Sepharose 4B. The agglutinins showed multiple species and were composed of eight heterogeneous subunits with molecular weights of 45,000, 42,000, 41,000, 39,000, 33,000, 29,000, 27,000, and 22,000 as determined by SDS-polyacrylamide gel electrophoresis. The affinity-isolated agglutinins were fractionated into four groups by gel filtration on a Fractogel TSK (Toyopearl) HW-65 column, and these were designated as Tachypleus tridentatus agglutinin (TTA)-I, -II, -III, and -IV in the order of elution. These agglutinins were demonstrated to be heterogeneous as judged by their specificity towards horse erythrocytes, subunit structures, and immunological properties. TTA-III showed a potent agglutination activity towards horse erythrocytes and was further purified by gel filtration on a Cellulofine GC-700 column. The purified TTA-III is a highly purified (46,000-fold) protein composed of homogeneous subunits (Mr, 42,000) as judged by SDS-polyacrylamide gel electrophoresis and immunological analysis.

Animals↗
PubMed1982

Studies on hemolymph nodes. I. Histology of the renal hemolymph node of the rat.

The structure of the left renal hemolymph node of young adult male Swiss albino rats has been studied by optical microscopy of semithin plastic embedded sections. Afferent lymphatics were found, opening into the subcapsular sinus through valved ostia, in a large majority of nodes. Free erythrocytes were found in the afferent lymphatics and in the subcapsular sinus. In the intermediary and medullary sinuses, erythrocytes were mostly attached to sinus macrophages. The efferent lymphatics contained few erythrocytes. All the sinuses macrophages. The efferent lymphatics contained few erythrocytes. All the sinuses contained a stained precipitate of lymph protein. The node is hypoactive in its general form but does not differ essentially from typical lymph nodes, except that it receives, through its afferent lymphatics, lymph containing numerous erythrocytes.

Animals↗
PubMed1982

Studies on hemolymph nodes. III. Renal lymph as a major source of erythrocytes in the renal hemolymph node of rats.

The content of erythrocytes in the intrinsic lymphatics of the kidneys of young adult male Swiss-Albino rats was studied by optical and transmission electron microscopy, in material fixed by vascular perfusion. Erythrocytes were consistently found in lymphatics associated with interlobular, arcuate, interlobar and hilar arteries. They are presumed to have entered lymphatics by diapedesis through peritubular capillaries and veins with which they are frequently in intimate contact. The findings confirm the view that, in the rat, the kidney is an important source of the erythrocytes found in the renal hemolymph node.

Animals↗
PubMed1993

Effects of parasitism by the braconid wasp Cotesia congregata on host hemolymph proteins of the tobacco hornworm, Manduca sexta.

Parasitism by the braconid wasp Cotesia congregata causes major alterations in the hemolymph proteins of host tobacco hornworm larvae. Earlier studies showed that the total amount of hemolymph protein is reduced during parasitism, beginning almost immediately after the host is parasitized. Simultaneously, parasitism induces synthesis of large amounts of novel proteins that appear in the blood as early as 1-2 h post-parasitization. The present report confirms earlier studies describing the presence of novel proteins in last instar hosts, and also characterizes the effects of parasitism in altering the titers of several endogenous host hemolymph proteins normally produced by the fat body and other tissues. Analysis of hemolymph plasma using SDS-PAGE and densitometry, as well as immunodiffusion assays, showed that in terminal stage fifth instar host larvae, the titers of serpins and arylphorin were dramatically reduced relative to the levels of these proteins detected in nonparasitized gate II fifth instar larvae of the same age. The relative differences between parasitized and nonparasitized larvae increased with time following ecdysis to the fifth instar, so that the day 4 nonparasitized larvae had arylphorin titers of c. 30 mg/ml, whereas parasitized day 4 larvae with newly emerged wasps had only one sixth that amount of storage protein circulating in the hemolymph. Similarly, in nonparasitized larvae the hemolymph serpin concentration increased from c. 200 micrograms/ml (on day 0) to > 600 micrograms/ml (on day 4) in prewandering gate II larvae, but in parasitized larvae the hemolymph serpin concentration was maintained in the range of 100-200 micrograms per ml hemolymph until the pharate third instar parasites emerged from the host larva on day 4. In contrast, the level of hemolymph lipophorin was unaffected by parasitism, and lipophorin increased from c. 1.3 to > 3 mg/ml during the time interval between days 0 and 4 in both nonparasitized and parasitized larvae. Hemolymph titers of insecticyanin also were not significantly different in parasitized vs nonparasitized larvae, and in both types of larvae the concentration of this pigment decreased by c. 50% during the same time interval when lipophorin was increased significantly. Instead of causing a generalized inhibition of host hemolymph protein synthesis, parasitism causes a complex array of changes in the hemolymph protein profile of Manduca sexta, possibly via the mediation of hormonal modulators of host protein synthesis, or transcriptional or translational regulation of host gene expression by factors associated with the polydnavirus or molecules secreted by the parasites.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Indigenous bacteria in hemolymph and tissues of marine bivalves at low temperatures.

Hemolymph and soft tissues of Pacific oysters (Crassostrea gigas) kept in sand-filtered seawater at temperatures between 1 and 8 degrees C were normally found to contain bacteria, with viable counts (CFU) in hemolymph in the range 1.4 x 10 to 5.6 x 10 bacteria per ml. Pseudomonas, Alteromonas, Vibrio, and Aeromonas organisms dominated, with a smaller variety of morphologically different unidentified strains. Hemolymph and soft tissues of horse mussels (Modiolus modiolus), locally collected from a 6- to 10-m depth in the sea at temperatures between 4 and 6 degrees C, also contained bacteria. The CFU in horse mussel hemolymph was of the same magnitude as that in oysters (mean, 2.6 x 10), and the bacterial flora was dominated by Pseudomonas (61.3%), Vibrio (27.0%), and Aeromonas (11.7%) organisms. In soft tissues of horse mussels, a mean CFU of 2.9 x 10 bacteria per g was found, with Vibrio (38.5%), Pseudomonas (33.0%), and Aeromonas (28.5%) constituting the major genera. After the challenge of oysters in seawater at 4 degrees C to the psychrotrophic fish pathogen Vibrio salmonicida (strains NCIMB 2245 from Scotland and TEO 84001 from Norway) and a commensal Aeromonas sp. isolated from oysters, the viable count in hemolymph increased 1,000-fold to about 10 bacteria per ml. In soft tissues, about a 1,000-fold increase in CFU to 6 x 10 was observed. V. salmonicida NCIMB 2245 invaded hemolymph and soft tissues after 14 days and dominated these compartments after 41 days, whereas strain TEO 84001 did not invade soft tissues to the same extent. Challenge with V. salmonicida NCIMB 2245 resulted in 100% mortality, whereas about 50% of the oysters survived challenge with the Norwegian strain, TEO 84001. The commensal Aeromonas sp. invaded hemolymph and soft tissues and caused 100% mortality. Oyster hemolymph contained agglutinins for Vibrio anguillarum but not for V. salmonicida, whereas we did not find agglutinins for either of these bacteria in horse mussels. Agglutinins for horse and human erythrocytes were found in hemolymph from both animals. We found no differences in agglutinin titers in oysters from different Norwegian locations, and long-term challenge with bacteria in seawater did not result in changes of agglutinin activity. These studies demonstrate that bacteria exist in hemolymph and soft tissues of marine bivalves at temperatures below 8 degrees C. Increased bacterial numbers in seawater at 4 degrees C result in augmented invasion of bacteria in hemolymph and soft tissues. V. salmonicida, a bacterium pathogenic for fish at low temperatures, invades bivalve hemolymph and soft tissues, and thus bivalves may serve as a reservoir for pathogens of fish at low seawater temperatures.

Journal Article↗
PubMedDate not supplied

Enhancement of Sf-9 cell growth and longevity through supplementation of culture medium with hemolymph.

The benefits of insect cell culture medium supplementation with hemolymph of Lonomia obliqua were investigated. The addition of hemolymph to the medium induced high levels of cell growth, and the viability was maintained for longer periods. The maximum cell yield increased almost 3-fold after hemolymph supplementation. Cultures in their stationary phase were rescued through hemolymph supplementation, also reaching high cell concentrations. These actions were much dependent on the concentration of hemolymph; low hemolymph concentration had a positive effect in cell growth, whereas high hemolymph concentration showed a deleterious effect. Fractionation of hemolymph by gel filtration chromatography showed the presence of three factors with different activity in insect cell culture: an potential anti-apoptotic factor, a growth-promoting factor, and an enzyme that hydrolyzes sucrose. Addition of hemolymph to the medium induced high levels of glucose production. The sucrose to glucose conversion was also linearly dependent upon the hemolymph concentration. Therefore, we conclude that cell growth and longevity can be increased by supplementation of the culture medium with hemolymph.

Animals↗
PubMed2000

Host plant urease in the hemolymph of the silkworm, Bombyx mori.

Urease activity was detected in the hemolymph of the silkworm, Bombyx mori from the beginning of spinning to the pharate adult stage if the larvae were reared on mulberry leaves throughout the 5th-instar (the last larval instar). In contrast, no urease activity was detected in the hemolymph of insects fed artificial diets, resulting in accumulation of urea during the spinning stage. To identify the hemolymph urease, the enzyme was highly purified from the hemolymph of the spinning larvae that had been reared on mulberry leaves and the properties of the purified enzyme were compared with those of the mulberry leaf urease. Four out of six monoclonal antibodies raised against jack bean seed urease cross-reacted equally with the silkworm hemolymph urease and the mulberry leaf urease. Under reducing conditions, the hemolymph urease and the mulberry leaf urease migrated at 90.5 kDa on SDS-PAGE gels. The first 20 N-terminal sequence of the hemolymph urease revealed complete identity with that of the leaf urease. The optimum pH for activity and Km value for urea were almost the same for the two enzymes. In conclusion, these two ureases are very likely identical, strongly suggesting that the mulberry leaf urease passes through the larval gut wall into the hemolymph without being digested. In addition, oral administration of mulberry leaf urease just before spinning induced considerable urease activity in the hemolymph of the larvae, but the same treatment did not induce enzyme activity in the hemolymph of the larvae three days before the onset of spinning. These results suggest that the silkworm larvae acquire the host plant urease specifically at the end of the feeding stage in order to degrade urea accumulated in the hemolymph.

Journal Article↗
PubMed2005

Reference ranges for hemolymph chemistries from Elliptio complanata of North Carolina.

Hemolymph chemistries may be useful nonlethal measures of bivalve health. The prognostic value of hemolymph, however, depends on a comparison of chemistry results to reference ranges from healthy individuals. Currently, knowledge of expected hemolymph values in healthy and unhealthy freshwater mussels is extremely limited. The purpose of this study was to develop a set of reference ranges for clinical evaluation of hemolymph from a freshwater mussel species common to southeastern USA. We collected hemolymph from 380 Elliptio complanata from 19 apparently healthy populations from northwest of Raleigh, North Carolina, during May through July 2001. We present reference ranges for hemolymph parameters ammonia, glucose, calcium, magnesium, phosphorus, aspartate aminotransferase (AST), bicarbonate, protein and cell count, and for tissue glycogen. We compare the subpopulations of mussels from regions with an agricultural riparian buffer to those surrounded predominantly by forested lands. We further present correlations noted between hemolymph chemistries and physical or physiologic parameters. The only statistically significant differences between populations contiguous to agricultural and forested lands were in hemolymph calcium and glucose concentrations. Other statistically significant correlations identified were between gravidity and hemolymph protein concentration and tissue glycogen content, as well as between gravidity and parasite burden, and between shell length and hemolymph glucose, AST, calcium and bicarbonate concentrations. The results of this study will aid the interpretation of health measures from populations of E. complanata of similar geographic and seasonal origin.

Animals↗
PubMed1999

Levels of serotonin in the hemolymph of Aplysia are modulated by light/dark cycles and sensitization training.

Serotonin (5-hydroxytryptamine, 5-HT) modulates the behavior and physiology of both vertebrate and invertebrate animals. Effects of injections of 5-HT and the morphology of the serotonergic system of Aplysia indicate that 5-HT may have a humoral, in addition to a neurotransmitter, role. To study possible humoral roles of 5-HT, we measured 5-HT in the hemolymph. The concentration of 5-HT in the hemolymph was approximately 18 nM, a value close to previously reported thresholds for eliciting physiological responses. The concentration of 5-HT in the hemolymph expressed a diurnal rhythm. In addition, electrical stimulation that leads to long-term sensitization significantly increased levels of 5-HT in the hemolymph during training, 1.5 hr after training, and 24 hr after training. Moreover, levels of 5-HT in the hemolymph were significantly correlated with the magnitude of sensitization. The half-life of an increase in 5-HT in the hemolymph was approximately 0.5 hr. Therefore, the persistent increase of 5-HT in the hemolymph 24 hr after sensitization training indicates that training caused a long-lasting increase in the release of 5-HT. This long-lasting increase in 5-HT in the hemolymph was blocked by treatment with an inhibitor of protein synthesis during training. Based on the levels of 5-HT in the hemolymph and its regulation by environmental events, we propose that 5-HT has a humoral role in regulation of the behavioral state of Aplysia. In support of this hypothesis, we found that increasing levels of 5-HT in the hemolymph led to significant alterations in feeding behavior. Increasing levels of 5-HT during the daytime when they were normally low increased the latency to assume feeding posture from daytime to nighttime values.

Animals↗
PubMed2001

Survival and differential development of Entomophaga maimaiga and Entomophaga aulicae (Zygomycetes: Entomophthorales) in Lymantria dispar hemolymph.

The closely related entomophthoralean fungi Entomophaga aulicae and E. maimaiga are both host-specific pathogens of lepidopteran larvae. However, these fungi do not have the same host range. The first objective of this study was to compare the fate of E. aulicae in the nonpermissive host Lymantria dispar with the fate of the successful pathogen E. maimaiga over the same time period. In the hemolymph of L. dispar injected with E. maimaiga protoplasts, the number of hemocytes demonstrated a decreasing trend after the first day postinjection and hemocytes completely disappeared by day 5, with the majority of larvae dying in 5.6 +/- 0.1 days. In L. dispar larvae, E. maimaiga infections developed successfully, evidenced by increasing numbers of protoplasts and hyphal bodies prior to host mortality. In contrast, at day 5 hemocytes were readily visible in hemolymph of E. aulicae-injected larvae, but E. aulicae cells did not increase in numbers, although persisting in the hemolymph for at least 16 days postinjection. For both fungal species, when hemolymph samples from injected insects were introduced to culture media viable fungal cultures were always produced. Both E. aulicae and E. maimaiga occurred in hemolymph initially after injection as protoplasts. For E. maimaiga, after day 3, <50% of fungal cells were hyphal bodies until insect death when most cells regenerated cell walls. For E. aulicae, from day 2 equal numbers of fungal cells in the hemolymph occurred as protoplasts and hyphal bodies. To investigate the cause of fungistasis in E. aulicae-injected larvae, E. aulicae cell cultures exposed to partially purified protein fractions from hemolymph of larvae infected with either fungus displayed increased lysis and decreased viability at lower concentrations of protein fractions compared with E. maimaiga cell cultures. These studies demonstrate that E. aulicae does not increase in L. dispar hemolymph, although it persists and results suggest that proteinaceous factors induced within the hemolymph may limit the capacity of E. aulicae to develop successful infections.

Animals↗
PubMed1989

Insect hemolymph factor promotes muscle histolysis in Solenopsis.

Hemolymph was collected from both normal (virgin) females (control hemolymph) and artificially inseminated females (experimental hemolymph) of Solenopsis, ssp., the imported fire ant (primarily S. invicta and S. geminata). When the control hemolymph was injected into normal females, breakdown of the thoracic flight musculature was not seen 24 hr postinjection. In contrast, when the experimental hemolymph was injected into normal females, flight muscle histolysis was marked 24 hr postinjection. When the experimental hemolymph was heated to 70 degrees C prior to injection into normal females, subsequent flight muscle breakdown was not seen. The injection of freshly collected semen into normal females produced no effect on flight muscle structure. Also, the injection of the experimental hemolymph into normal, alate males produced no muscle histolysis. These observations suggest that the hemolymph from inseminated females contains a factor (or factors) that induces the specific breakdown and subsequent dissolution of the complex thoracic flight musculature. Other thoracic muscles (leg muscles, intersegmental muscles, etc.) are not affected. Based on observations made in both insect muscles as well as vertebrate skeletal muscles, it is suggested that such a hemolymph factor may act by disrupting the structural integrity of muscle cell membranes, resulting in significant changes in membrane permeability, especially with regard to calcium ions.

Animals↗
PubMed1999

Lipophorin of female Blattella germanica (L.): characterization and relation to hemolymph titers of juvenile hormone and hydrocarbons.

High density lipophorin (HDLp) from the hemolymph of the German cockroach, Blattella germanica (L.) (Family Blattellidae), has an apparent molecular weight of 670kDa, with an isoelectric point of 7.0 and a density of 1.109g/ml. It is composed of two subunits, apolipoprotein-I (212kDa) and apolipoprotein-II (80kDa), and consists of 51.4% lipid, 46.2% protein and 2.4% carbohydrate. Hydrocarbons constitute 42.2% of the total lipids which also contain diacylglycerol, cholesterol and phospholipid. Lipophorin is rich in the amino acids glutamic acid, aspartic acid, lysine, valine, and leucine. Specificity of a polyclonal antibody was demonstrated by Western blotting and Ouchterlony immunodiffusion: the antiserum recognized native HDLp and apolipoprotein-I, but not apolipoprotein-II, purified vitellin, or other hemolymph proteins. It also recognized a protein in the hemolymph of Supella longipalpa (Blattellidae) but did not cross-react with hemolymph proteins from Periplaneta americana (Blattidae) or Diploptera punctata (Blaberidae). An enzyme-linked immunosorbent assay was developed to measure the HDLp titer in the hemolymph of adult females. The titer of HDLp, a juvenile hormone binding protein, exhibited no clear relationship to the changing titer of juvenile hormone in hemolymph. The hemolymph titer of hydrocarbon, which is also carried by HDLp, showed some functional relation to the concentration of HDLp in the hemolymph. Because it concurrently serves multiple functions in insect development and reproduction, lipophorin titer might covary with the titers of lipid ligands that occur at high concentrations and require extensive shuttling through the hemolymph.

Journal Article↗
PubMed2003

Hemolymph ecdysteroids do not affect vitellogenesis in the lubber grasshopper.

The role of hemolymph ecdysteroids in the reproduction of non-dipteran insects is unclear. We examine the role(s) of hemolymph ecdysteroids during egg production in the lubber grasshopper, Romalea microptera. In all individuals, hemolymph ecdysteroids rose to a sharp peak with similar maxima and then fell to undetectable levels. The time from the adult molt to the maximum ecdysteroid titer (E(max) titer) varied in response to food availability, whereas the time from E(max) titer to oviposition was unrelated to food availability. Because both the timing of egg production and the timing of E(max) responded similarly to environmental changes, ecdysteroids may be involved in egg production. We hypothesized that this role is the stimulation of vitellogenesis. Ovariectomized females had vitellogenin but no ecdysteroids, so ecdysteroids are not necessary for vitellogenin production. In addition, treatment of females with ecdysteroids altered neither Vg titers nor ovarian growth. Ovarian ecdysteriods increased at the same age in development as hemolymph ecdysteroids. In contrast to hemolymph ecdysteroids, ovarian ecdysteroids persisted until oviposition. Despite this, [(3)H]ecdysone injected into the hemolymph was detected later only at very low levels in the ovary, suggesting that hemolymph ecdysteroids are not sequestered by the ovary. In summary, our studies indicate that hemolymph ecdysteroids in adult females of the lubber grasshopper are associated with the timing of egg production, but they neither regulate vitellogenesis nor act as a source of ecdysteroids for the ovary.

Analysis of Variance↗
PubMed2000

Catechol concentrations in the hemolymph of the scallop, Placopecten magellanicus.

Catecholamines have previously been detected in numerous tissues and are thought to control a wide variety of physiological functions in bivalve molluscs. In the present study, alumina extraction and high-performance liquid chromatography reveal the presence of significant concentrations of 3,4-dihydroxyphenylalanine (DOPA), dopamine, and 3,4-dihydroxyphenylacetic acid (DOPAC) in the hemolymph of the sea scallop, Placopecten magellanicus. The concentration of dopamine in the hemolymph averaged 223.8 ng/ml, (+/-48.4, SEM), equivalent to 10(-7) to 10(-6) M. Neither epinephrine nor norepinephrine was reliably detected in significant quantities. Previous studies have demonstrated physiological responses to dopamine with thresholds of 10(-9) to 10(-6) M, thus suggesting that this catecholamine may have an endocrine function. Furthermore, monitoring hemolymph concentrations of catechols might provide a sensitive measure of the physiological status of bivalves. For example, drugs known to affect catechol concentrations in other tissues also effect hemolymph levels. Administration of monoamine oxidase inhibitors such as pargyline, deprenyl, and clorgyline at 10(-4) M for 1 day of incubation followed by a 2-day wash resulted in decreased hemolymph concentrations of DOPAC and increased concentrations of its precursors, DOPA and dopamine. Incubation in 10(-4) M 3,5-dinitrocatechol, a catecholamine-O-methyl transferase blocker, for 1 day followed by a 2-day wash significantly increased the concentration of dopamine and DOPAC in the hemolymph. Scallops incubated in 10(-5) M alpha-methyl-p-tyrosine, a blocker of tyrosine hydroxylase, for 1 day followed by a 3-day wash in artificial seawater had significantly reduced concentrations of DOPA, dopamine, and DOPAC in the hemolymph. In addition to responding to pharmacological agents, dopamine levels also decreased significantly following thermal induction of spawning, thus suggesting that hemolymph concentrations of catechols might provide indices of reproductive activity and/or stress.

3,4-Dihydroxyphenylacetic Acid↗