Guibbolini Marielle, Borelli Gil, Mayer-Gostan Nicole, Priouzeau Fabrice, De Pontual Hélène, Allemand Denis, Payan Patrick
UMR INRA-UNSA N 1112, Laboratoire ROSE, Université de Nice-Sophia Antipolis, Faculté des Sciences, Parc Valrose, 06108 Nice Cedex 2, France.
Comp Biochem Physiol A Mol Integr Physiol. 2006 Sep;145(1):99-107. doi: 10.1016/j.cbpa.2006.05.003. Epub 2006 May 10.
The aim of the present work was to examine the modifications of the organic composition of fish endolymph under environmental conditions (day-night cycle, starvation and Cl2-stress) known to modify otolith growth. Endolymph electrophoretic patterns were compared. An antibody raised against the trout otolith organic matrix allowed examining the variations of organic matrix precursors in the endolymph under the above conditions. Western blot analysis showed bands around 60-80 kDa. A 50% decrease of immunolabelling was observed during the night whereas increases were seen after starvation (factor 3) or stress (factor 2) suggesting that these variations could be related to the organic matrix deposit. A factor retarding in vitro CaCO3 crystallization (FRC) was shown to co-precipitate with endolymph proteins and its apparent molecular mass (determined by measuring the activity after electro elution of gel electrophoresis) was estimated around 20 kDa. The FRC activity was stable during day-night cycle whereas it decreased by 70% and nearly 100% under starvation and stress respectively. These results suggest that the FRC, although retarding in vitro crystallization, plays a major role in the process of otolith calcification and that the decreases measured after starvation and stress are responsible for the decreases of the otolith growth. The variations of these two parameters (precursors and FRC) could contribute for the changes in the microstructure of the otolith.
本研究的目的是在已知会改变耳石生长的环境条件(昼夜循环、饥饿和氯气胁迫)下,研究鱼类内淋巴有机成分的变化。对内淋巴电泳图谱进行了比较。一种针对鳟鱼耳石有机基质产生的抗体,用于检测上述条件下内淋巴中有机基质前体的变化。蛋白质免疫印迹分析显示在60 - 80 kDa附近有条带。夜间观察到免疫标记减少了50%,而饥饿(3倍因子)或应激(2倍因子)后则增加,这表明这些变化可能与有机基质沉积有关。一种体外碳酸钙结晶延迟因子(FRC)被证明与内淋巴蛋白共沉淀,其表观分子量(通过测量凝胶电泳电洗脱后的活性来确定)估计约为20 kDa。FRC活性在昼夜循环中保持稳定,而在饥饿和应激条件下分别下降了70%和近100%。这些结果表明,FRC虽然在体外延迟结晶,但在耳石钙化过程中起主要作用,饥饿和应激后测量到的其活性下降是耳石生长下降的原因。这两个参数(前体和FRC)的变化可能导致耳石微观结构的改变。