Gill ion transport and copper accumulation in the blue crab Callinectes sapidus

Paganini, Christianne Lorea; Souza, Maurício de; Bianchini, Adalto

doi:10.1016/j.cbpc.2008.10.049

Cited by 3 publications

(11 citation statements)

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“…Experimental media were completely renewed every 24 h. At the beginning and after 24 h of exposure, nonfiltered and filtered (0.45-mm-mesh filter) water samples were collected from the experimental media for further analyses of total (nonfiltered samples) and dissolved (filtered samples) Cu concentrations by AAS (Avanta 932 Plus; GBC), as previously described [12,13,23]. Six blue crabs were tested under each experimental condition.…”

Section: Experimental Animals and Cu Exposurementioning

confidence: 99%

“…Copper was added to the experimental media from a stock solution (5 mM). Copper concentration was checked by atomic absorption spectrophotometry (AAS; Avanta 932 Plus; GBC) using standard solutions prepared in saltwater at the experimental salinity [12,13,23]. Copper was added to the water 3 h prior to crab introduction in the test chamber to allow Cu equilibration with the experimental medium [23].…”

Section: Experimental Animals and Cu Exposurementioning

confidence: 99%

“…, are involved in Cu transport across the gill epithelium [11,13,16]. Therefore, Na þ transport across the gills can be affected by the presence of waterborne Cu, resulting in osmoregulatory disturbances.…”

Section: Introductionmentioning

confidence: 99%

“…This metal can compete with other ions such as Na þ [10,13] and Ca 2þ [14,15] for binding physiologically active sites at the gill, consequently being incorporated by the animal. In fact, components of Na þ transport, including the enzymes sodium-potassium adenosine triphosphatase (Na þ /K þ -ATPase), hydrogen adenosine triphosphatase (H þ -ATPase), and carbonic anhydrase (CA), as well as the membrane sodium-hydrogen exchanger (Na þ /H þ ) and sodium-potassium-chloride cotransporter (Na…”

Section: Introductionmentioning

confidence: 99%

“…Physiological studies have demonstrated that the key mechanism of waterborne Cu toxicity is associated with an impairment of Na þ and Cl À regulation in both freshwater and marine osmoregulating fish and invertebrates. Generally, the gill is the main organ involved in osmotic and ionic regulation and is consequently the main target for Cu toxicity in these animals [10,11,13].…”

Section: Introductionmentioning

confidence: 99%

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mRNA Expression and activity of ion‐transporting proteins in gills of the blue crab Callinectes sapidus: Effects of waterborne copper

Martins

Almeida

Marins

et al. 2010

Enviro Toxic and Chemistry

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Waterborne Cu effects on the transcription of genes encoding ion-transporting proteins and the activities of these proteins were evaluated in gills of the blue crab Callinectes sapidus acclimated to diluted (2‰) and full (30‰) seawater. Crabs were exposed (96 h) to an environmentally relevant concentration of dissolved Cu (0.78 µM) and had their posterior (osmoregulating) gills dissected for enzymatic and molecular analysis. Endpoints analyzed were the activity of key enzymes involved in crab osmoregulation (sodium-potassium adenosine triphosphatase [Na(+)/K(+)-ATPase], hydrogen adenosine triphosphatase [H(+)-ATPase], and carbonic anhydrase [CA]) and the mRNA expression of genes encoding these enzymes and the sodium-potassium-chloride (Na(+)/K(+)/2Cl⁻) cotransporter. Copper effects were observed only in crabs acclimated to diluted seawater (hyperosmoregulating crabs) and were associated with an inhibition of the expression of mRNA of genes encoding the Na(+)/K(+)-ATPase and the Na(+)/K(+)/2Cl⁻ cotransporter. However, Cu did not affect Na(+)/K(+)-ATPase activity, indicating that the gene transcription is downregulated before a significant inhibition of the enzyme activity can be observed. This also suggests the existence of a compensatory response of this enzyme to prevent osmoregulatory disturbances after short-term exposure to environmentally relevant Cu concentrations. These findings suggest that Cu is a potential ionoregulatory toxicant in blue crabs C. sapidus acclimated to low salinity. The lack of Cu effect on blue crabs acclimated to full seawater would be due to the reduced ion uptake needed for the regulation of the hemolymph osmotic concentration in full seawater (30‰). Also, this could be explained considering the lower bioavailability of toxic Cu (free ion) associated with the higher ionic content and dissolved organic matter concentration in high salinity (30‰) than in diluted seawater (2‰).

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Section: Experimental Animals and Cu Exposurementioning

confidence: 99%

Section: Experimental Animals and Cu Exposurementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

mRNA Expression and activity of ion‐transporting proteins in gills of the blue crab Callinectes sapidus: Effects of waterborne copper

Martins

Almeida

Marins

et al. 2010

Enviro Toxic and Chemistry

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Mechanisms of copper accumulation in isolated mantle cells of the marine clam Mesodesma mactroides

Lopes

Barcarolli

Oliveira

et al. 2011

Enviro Toxic and Chemistry

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In vivo copper accumulation was determined in tissues (mantle, gills, digestive gland, and hemolymph) following exposure to Cu (5 µM) for up to 96 h. Mantle was the tissue that accumulated the most Cu, followed by gill, digestive gland, and hemolymph. Therefore, in vitro Cu accumulation was evaluated in isolated mantle cells exposed to 0.5, 1.0, 2.5, and 5.0 µM Cu for 1 and 3 h. After both exposure times, no change in cell viability was observed. However, a significant Cu accumulation was observed in cells exposed to 2.5 and 5.0 µM Cu. Cell exposure to 2.5 µM Cu for 1 h did not affect the ionic (Na(+), K(+), Ca(2+), and Cl(-)) content of isolated mantle cells, characterizing an "ideal" noneffect concentration for the study of the involvement of different ion-transporting proteins (Na(+), K(+), and Cl(-) channels; Na(+)/K(+) 2Cl(-) and Na(+)/Cl(-) cotransporters; Na(+)/Ca(2+), Cl(-)/HCO3-, and Na(+)/H(+) exchangers; Na(+)/K(+) -ATPase; V-ATPase; and carbonic anhydrase) in Cu accumulation. Isolated cells were pre-exposed (30 min) to specific blockers or inhibitors of the ion-transporting proteins and then exposed (1 h) to Cu (2.5 µM) in the presence of the drug. A significant increase of 29.1 and 24.3% in Cu accumulation was observed after cell incubation with acetozalamide (carbonic anhydrase inhibitor) and NPPB (Cl(-) channels blocker), respectively. On the other hand, a significant decrease (48.2%) in Cu accumulation was observed after incubation with furosemide (Na(+) /K(+)/2Cl(-) blocker). Taken together, these findings indicate the mantle as an important route of Cu entry in M. mactroides, pointing to the cotransporter Na(+)/K(+)/2Cl(-) as a major mechanism of Cu accumulation in mantle cells of the clam.

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Acute toxicity, accumulation and tissue distribution of copper in the blue crab Callinectes sapidus acclimated to different salinities: In vivo and in vitro studies

et al. 2011

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In vivo and in vitro studies were performed to evaluate acute toxicity, organ-specific distribution, and tissue accumulation of copper in Callinectes sapidus acclimated to two different experimental salinities (2 and 30 ppt). Blue crabs were quite tolerant to copper. Acute dissolved copper toxicity (96-h LC(50) and its corresponding 95% confident interval) was higher at salinity 2 ppt (5.3 (3.50-8.05) μM Cu) than at 30 ppt (53.0 (27.39-102.52) μM Cu). The difference between salinities can be completely explained based on the water chemistry because it disappeared when 96-h LC(50) values were expressed as the free Cu(2+) ion (3.1 (1.93-4.95) μM free Cu at 2 ppt versus 5.6 (2.33-13.37) μM free Cu at 30 ppt) or the Cu(2+) activity (1.4 (0.88-2.26) μM Cu activity at 2 ppt versus 1.7 (0.71-4.07) μM Cu activity at 30 ppt). The relationships between gill Cu burden and % mortality were very similar at 2 and 30 ppt, in accord with the Biotic Ligand Model. In vivo experiments showed that copper concentration in the hemolymph is not dependent on metal concentration in the surrounding medium at either experimental salinity. They also showed that copper flux into the gills is higher than into other tissues analyzed, and that anterior and posterior gills are similarly important sites of copper accumulation at both experimental salinities. In vitro experiments with isolated-perfused gills showed that there is a positive relationship between copper accumulation in this tissue and the metal concentration in the incubation media for both anterior and posterior gills. A similar result was observed at both low and high salinities. Furthermore, in vitro experiments showed that copper accumulation in posterior gills is also positively and strongly dependent on the incubation time with copper. Gill copper accumulation occurred at a lower rate in the first 2h of metal exposure, increasing markedly after this "steady-state" period. This finding was corroborated by a significant increase in copper influx to the gill perfusate (corresponding to crab hemolymph) after this time, measured using (64)Cu. In vivo, after uptake from solution, (64)Cu was primarily accumulated in the gills and the rest of the body rather than in the hemolymph, hepatopancreas, or other internal tissues. Overall, the present findings indicate that gills are a key target organ for copper accumulation, as well as an important biological barrier against the excessive uptake of copper into the hemolymph and the subsequent distribution of this metal to internal organs of the blue crab.

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Gill ion transport and copper accumulation in the blue crab Callinectes sapidus

Cited by 3 publications

References 0 publications

mRNA Expression and activity of ion‐transporting proteins in gills of the blue crab Callinectes sapidus: Effects of waterborne copper

mRNA Expression and activity of ion‐transporting proteins in gills of the blue crab Callinectes sapidus: Effects of waterborne copper

Mechanisms of copper accumulation in isolated mantle cells of the marine clam Mesodesma mactroides

Acute toxicity, accumulation and tissue distribution of copper in the blue crab Callinectes sapidus acclimated to different salinities: In vivo and in vitro studies

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