Induction of mRNAs in response to acclimation of trout cells to different osmolalities |
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Authors: | T Sakamoto N Ojima M Yamashita |
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Institution: | (1) National Research Institute of Fisheries Science, Yokohama, 236-0004, Japan;(2) Faculty of Integrated Arts and Sciences, Hiroshima University, Higashi-hiroshima, 739-8521, Japan |
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Abstract: | The direct effect of osmolality on growth and mRNA population were investigated in the rainbow trout cell line (RTG-2). These cells can grow in the media of osmolalities ranging from 200 to 600 mosmol kg-1. With two-dimensional electrophoresis, the in vitro translation of poly(A+) RNA isolated from these cells showed osmoresponsive changes in the population of translatable mRNAs. Using differential mRNA display polymerase chain reaction, however, we identified inducible cDNA products in hyper-osmotic and hypo-osmotic media as third component of complement, and as homologues of known genes: an atypical protein kinase regulated by the thyrotropin-dependent mitogenic pathway, nucleolin and CHD3. The remaining cDNAs have no significant homology in GenBank. Northern blots demonstrate that their mRNA levels were induced in hyper-osmotic and hypo-osmotic media, but not by other stresses. The expressed proteins of these mRNAs may be involved directly or indirectly in the adaptation of RTG-2 cells to different osmolalities probably through the osmotic signal transduction and adjustment in cellular metabolism to osmotic stress. |
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Keywords: | CHD3 cell culture complement differential mRNA display polymerase chain reaction growth nucleolin protein kinase rainbow trout RTG-2 cell salinity |
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