The cloning of eel osmotic stress transcription factor and the regulation of its expression in primary gill cell culture

W. K.F. Tse, S. C. Chow, C. K.C. Wong

研究成果: Contribution to journalArticle査読

27 被引用数 (Scopus)

抄録

In the present study, we aimed to clone an osmotic stress transcriptional factor (Ostf) from gill cells of Japanese eels. In addition, we measured its expression in Percoll™-gradient-isolated gill chloride (CC) and pavement (PVC) cells and determined the regulation of its expression in primary gill cell culture. Using degenerative primers and RACE techniques, we cloned a cDNA of 615 bp, encompassing the coding sequence of Ostf (204 amino acids). The cloned Ostf1 DNA sequence shared 84% DNA homology with the Ostf1 of tilapia. In general, the basal Ostf expression level was found to be significantly higher in CCs than in PVCs. In the direct transfer of fish from freshwater to seawater, a significant but transient induction of Ostf mRNA in CCs and PVCs was measured after 6 h of acclimation. Compared with gill CCs, the level of induction measured at PVCs was lower. In the seawater-to-freshwater transfer, no significant change in Ostf transcript levels was detected in either CCs or PVCs. To decipher the regulatory mechanism of Ostf expression, we conducted experiments using primary gill cell culture to specifically address the involvement of two putative osmosensors (i.e. intracellular ion strength/macromolecular crowding and cytoskeleton) in the regulation of Ostf expression. Hypertonic treatment using impermeable solutes (i.e. NaCl, 500 mOsmol l-1) induced Ostf mRNA expression in 6 h, but no noticeable effect was measured using permeable solute (i.e. urea, 500 mOsmol l -1). The induction was transcriptionally regulated and was abolished by the addition of organic osmolytes (i.e. betaine, inositol or taurine) into the culture media. Addition of colchicine (an inhibitor of microtubule polymerization) to hypertonic (with added NaCl, 500 mOsmol l-1) cells reduced Ostf mRNA expression, suggesting that an increase in intracellular ionic strength and the integrity of the cytoskeleton are involved in the activation of Ostf mRNA expression in the cells. Collectively, the results of this study reveal, for the first time, the differential expression of Ostf in isolated CCs and PVCs. The resulting knowledge can shed light on how Ostf participates in hyperosmotic adaptation in fish gills.

本文言語英語
ページ(範囲)1964-1968
ページ数5
ジャーナルJournal of Experimental Biology
211
12
DOI
出版ステータス出版済み - 6 2008
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生態、進化、行動および分類学
  • 生理学
  • 水圏科学
  • 動物科学および動物学
  • 分子生物学
  • 昆虫科学

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