Biochemical Adaptation Lab, Department of Zoology, North Eastern Hill University, Shillong, 793 022, India.
Fish Physiol Biochem. 2010 Mar;36(1):17-27. doi: 10.1007/s10695-008-9275-5. Epub 2008 Nov 7.
The present study aimed at determining the effect of cell volume changes on protein synthesis, measured as the incorporation of [(3)H]leucine into acid-precipitable protein, in isolated hepatocytes of air-breathing walking catfish (Clarias batrachus). The rate of protein synthesis, which was recorded to be 10.02 +/- 0.10 (n = 25) nmoles mg(-1) cell protein h(-1) in isotonic incubation conditions, increased/decreased significantly by 18 and 48%, respectively, following hypo- (-80 mOsmol l(-1))/hypertonic (+80 mOsmol l(-1)) incubation conditions (adjusted with NaCl), with an accompanying increase/decrease of hepatic cell volume by 12 and 20%, respectively. Similar cell volume-sensitive changes of protein synthesis were also observed when the anisotonicity of incubation medium was adjusted with mannitol. Increase of hepatic cell volume by 9%, due to addition of glutamine plus glycine (5 mM each) to the isotonic control incubation medium, led to a significant increase of protein synthesis by 14%. Decrease of hepatic cell volume by 15 and 18%, due to addition of dibutyl-cAMP and adenosine in isotonic control incubation medium, led to a significant decrease of protein synthesis by 30 and 34%, respectively. Thus, it appears that the increase/decrease of hepatic cell volume, caused either by changing the extracellular osmolarity or by the presence of amino acids or certain other metabolites, leads to increase/decrease of protein synthesis, respectively, and shows a direct correction (r = 0.99) between the hepatic cell volume and protein synthesis in walking catfish. These cell volume-sensitive changes of protein synthesis probably help this walking catfish in fine tuning the different metabolic pathways for better adaptation during cell volume changes and also to avoid the adverse affects of osmotic stress. This is the first report of cell volume-sensitive changes of protein synthesis in hepatic cells of any teleosts.
本研究旨在确定细胞体积变化对蛋白质合成的影响,蛋白质合成通过 [(3)H]亮氨酸掺入酸沉淀蛋白来测量,在呼吸空气的步行鲶鱼(Clarias batrachus)的分离肝细胞中进行。在等渗孵育条件下,蛋白质合成的速率记录为 10.02 +/- 0.10(n = 25)nmoles mg(-1)细胞蛋白 h(-1),分别在低渗(-80 mOsmol l(-1))/高渗(+80 mOsmol l(-1))孵育条件下(用 NaCl 调节)显著增加/减少 18%和 48%,同时肝细 胞体积分别增加/减少 12%和 20%。当用甘露醇调节非等渗性孵育介质时,也观察到蛋白质合成的类似细胞体积敏感变化。由于向等渗对照孵育培养基中添加谷氨酰胺加甘氨酸(各 5 mM),肝细 胞体积增加 9%,导致蛋白质合成显著增加 14%。由于在等渗对照孵育培养基中添加二丁基-cAMP 和腺苷,肝细 胞体积分别减少 15%和 18%,导致蛋白质合成分别显著减少 30%和 34%。因此,似乎是细胞体积的增加/减少,无论是通过改变细胞外渗透压还是通过存在氨基酸或某些其他代谢物引起的,分别导致蛋白质合成的增加/减少,并显示在步行鲶鱼中肝细 胞体积和蛋白质合成之间存在直接校正(r = 0.99)。这些对蛋白质合成的细胞体积敏感变化可能有助于这种步行鲶鱼在细胞体积变化期间精细调整不同的代谢途径,以更好地适应,并避免渗透压应激的不利影响。这是首次报道任何硬骨鱼类肝细 胞中蛋白质合成的细胞体积敏感变化。