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Induction of mRNAs in response to acclimation of trout cells to different osmolalities
Authors:T Sakamoto  N Ojima  M Yamashita
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
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.
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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