Storey, K. Sokolova, I. Lesser, M. Vysotskaya, R. Lysenko, L.
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Folch, J. PubMed Google Scholar. Sidorov, V. Assays, Lososevye Salmonidae , vyp. Engelbrecht, F. Determination in serum: a rapid direction method, S. Tsyganov, E. Delo , , no. Arduini, A. Lipid Res. Alekseenko, L. Pokrovskii, A. Barrett, A.
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Kantserova, N. Bradford, M. Sukhovskaya, I. Habig, W. The first enzymatic step in mercapturic acid formation, J. The taurine transporter TAUT is related to the regulation of intracellular contents of taurine, the dominant osmolyte.
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These findings suggest that TAUT expression is regulated differently in gills and mantles following exposure to alterations in environmental salinity. Taken together, this study used the physiological, biochemical and molecular approaches to simultaneously explore the osmoregulation in tissues of hard clam and may further help to understand the osmoregulation in bivalves. Most marine mollusks are osmoconformers as the osmolality of the body fluids changes in the same direction as a change in environmental salinity Mantel and Farmer, It would be expected that osmoconformation would result in osmotic stress to tissues of marine mollusks following changes in surrounding salinity.
Systems that regulate both inorganic and organic osmolytes would be expected to be altered in response to changes in environmental salinity. Although NKA activity was found in a range of tissues in the clam, Rangia cuneat Saintsing and Towle, , the highest activity was found in the mantle.
NKA activity was also found to be highest in the mantle of the blue mussel, Mytilus galloprovincialis , while in another bivalve, Scapharca inasquivalvis , it was highest in the gills Borgatti et al. These findings indicate that the mantles and gills NKA may play an important role in the response to alterations in environmental salinity. In cellular osmoregulation, bivalves have the ability to mobilize organic osmolytes in addition to ion regulation as demonstrated in vertebrates Somero and Bowlus, Long-term accumulation of inorganic ions causes damage to cells, and as such, inorganic ions are not conducive to long-term osmoregulation in cells Yancey, In contrast, organic osmolytes are able to stabilize intracellular proteins without affecting cellular functions and suitable for the long-term accumulation in cells Yancey, ; Burg et al.
Hence, the modulation of intracellular organic osmolyte content is vital in multicellular organisms facing the changes in extracellular osmolality to maintain the cell volume Yancey, In bivalves, the content of free amino acid FAA was higher in the intracellular fluid than vertebrates Gilles, , Conversely, FAA was released to the hemolymph or extracellular fluid by modulating membrane and transporter properties upon hypoosmotic stress Bartberger and Pierce, ; Baginski and Pierce, ; Bishop et al.
Taurine is the dominant FAA and high intracellular taurine concentrations existed during osmoregulation in marine invertebrates Simpson et al. In hyperosmotic environments, extracellular taurine was transported into the cell via the taurine transporter TAUT for the maintenance of osmolality Wright and Secomb, ; Kwon and Handler, ; Burg et al. In reverse, taurine diffused out of the cells under hypoosmotic environments Fugelli and Riersen, ; Fugelli and Rohrs, The hard clam Meretrix lusoria is a marine inhabitant with extremely euryhaline behaviors throughout its life history.
However, the effects of changes in salinity on the cellular osmoregulatory mechanism of hard clam have not been investigated. It is required to explore whether these cellular osmoregulatory mechanisms work together in bivalves. By exploring these questions, it will help us to understand the mechanisms for the cellular osmoregulation of the hard clam. Hard clams, 30—50 mm and The hemolymph of the hard clam was collected using the 23 G syringe.
Photometric analysis was conducted using a Hitachi U spectrophotometer Tokyo, Japan. The gills and mantles were freshly dissected from the hard clams, and then were freeze-dried Osterode am Harz , germany for 10 h. After centrifugation, the supernatant was passed through a 0. The NKA activities of the gills and mantles were determined according to the method of Hwang et al. Homogenization was performed with a motorized Teflon pestle at rpm for 30 s. Next, the supernatant was used to determine protein concentrations and enzyme activities.
The inorganic phosphate concentration was measured according to Peterson's method Peterson, The enzyme activity of NKA was defined as the difference between the inorganic phosphate liberated in the presence and absence of 3. Each sample was assayed in triplicate. RNA integrity was verified by 0.
The occurrence of secondary products and primer-dimers was inspected using melting curve analysis and electrophoresis to confirm that the amplification was specific. One identical control sample was used as the internal control among different groups. The washed IP buffer was removed from the vial and preserved for the immunoblotting analysis.
The supernatant eluted sample from the vial was used for the immunoblotting analysis. In the present study, the pre-immune serum was also used to replace the TAUT antibody in the immunoblotting test for antibody specificity. The protein homogenate was prepared as described above, and was used for the immunoblotting. Results were converted to numerical values in order to compare relative intensities of immunoreactive bands.
N -value for the analyses of ions, osmolality and total amino acid is 8, and for the other analyses is 5. Free amino acids FAAs are important cellular osmolytes.
Salinity effects on concentrations of total free amino acids A and different free amino acids B in gills of the hard clam. Salinity effects on concentrations of total free amino acids A and different free amino acids B in mantles of the hard clam. A Full-length amino acid sequences of TAUT from different species were used to construct a phylogenetic tree by the neighbor-joiningmethod with bootstrap replicates.
Percentages of bootstrapping branch corrections are shown beside the branches, and the ortholog relationships of clades are labeled on the right side to indicate different subtree regions. The GenBank accession nos. The TAUT is expressed in the gill, mantle, muscle, heart, stomach, and intestine. Furthermore, expression levels were higher in the gill and mantle than in the other tissues examined. To further explore the role of TAUT in osmoregulation in the gill and mantle of hard calm, a specific antibody was derived to against hard clam TAUT protein.
Specificity analysis of the hard clam TAUT antibody. The arrow indicated the predicted immunoreactive band. B The pre-immune serum was used as the negative control, which revealed no immunoreactive band. The representative immunoblot for the protein expression was shown in B. Therefore, NKA activity may contribute to ion regulation in the mantle, but not in the gill during long-term hyperosmotic stress. Encyclopedia of Life Sciences. Australian Museum. Archived from the original on World Congress of Malacology.
Principles of Paleontology 2 ed. Freeman and Co. The Invertebrates, A Synthesis 3 ed. UK: Blackwell Science. December 22, Proceedings of the Royal Society B. BBC News. April Archived from the original on March 31, In Anderson, D. Invertebrate Zoology 2 ed. June 1, Bibcode : Sci Journal of the Geological Society. Bibcode : JGSoc. December Journal of Morphology.
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Journal of Shellfish Research. Athenaeus, The Deipnosophists. Cambridge, Massachusetts: Harvard University Press. Annual of the British School of Archaeology at Athens. Biblical Archaeologist. Merriam Co. June Annual Review of Anthropology. Archived from the original PDF on The Shell Money of the Slave Trade. Australian Venom Research Unit. UNSW Press. Drum and Croaker. Contact Dermatitis. The Festivus. New Scientist. September—October Revista da Sociedade Brasileira de Medicina Tropical.
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Ionic and Amino Acid Regulation in Hard Clam (Meretrix lusoria) in Response to Salinity Challenges
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