Deane E E, Kelly S P, Collins P M, Woo N Y S
Department of Biology, Chinese University of Hong Kong, Shatin, NT Hong Kong SAR, China.
Mar Biotechnol (NY). 2003 Jan-Feb;5(1):79-91. doi: 10.1007/s10126-002-0052-7.
To ascertain some of the important biochemical and molecular events that take place during early larval development of silver sea bream (Sparus sarba), we undertook a study of changes in the morphology as well as the ontogeny of the RNA-DNA ratio, growth hormone (GH), insulin-like growth factor I (IGF-I) messenger RNA abundance, Na(+)-K(+)-ATPase subunit mRNA abundance, and Na(+)-K(+)-ATPase enzyme activity. Larvae samples were collected at 1 to 46 days posthatch (dph). At 7 dph the yolk sac was fully absorbed, and from 28 dph onward larvae underwent rapid developmental changes to the juvenile stage. The RNA-DNA ratio was highest at 1 dph, decreased to low levels between 7 and 21 dph, then increased by 28 dph, and then again by 46 dph. The ontogenetic profiles of GH, IGF-I, and Na(+)-K(+)-ATPase alpha1 and beta1 subunits were studied using reverse transcriptase polymerase chain reaction, coupled with radioisotope hybridization of immobilized DNA. Growth hormone abundance reached a constant and high level from 35 dph onward, whereas the IGF-I level reached a peak at 35 dph and then significantly decreased. Both Na(+)-K(+)-ATPase alpha1 and beta1 subunit mRNAs increased up to 35 dph, however, at 46 dph the alpha1 subunit remained high whereas the beta1 subunit decreased. Na(+)-K(+)-ATPase activity was low in 1-dph larvae but increased rapidly as development progressed. The importance of these findings is discussed within the context of larval development.
为了确定在黑鲷幼鱼早期发育过程中发生的一些重要生化和分子事件,我们对黑鲷幼鱼的形态变化以及RNA-DNA比率、生长激素(GH)、胰岛素样生长因子I(IGF-I)信使RNA丰度、Na(+)-K(+)-ATP酶亚基mRNA丰度和Na(+)-K(+)-ATP酶活性的个体发生进行了研究。在孵化后1至46天(dph)收集幼鱼样本。在7 dph时卵黄囊完全吸收,从28 dph起幼鱼经历快速发育变化进入幼鱼期。RNA-DNA比率在1 dph时最高,在7至21 dph之间降至低水平,然后在28 dph时升高,随后在46 dph时再次升高。使用逆转录聚合酶链反应结合固定化DNA的放射性同位素杂交研究了GH、IGF-I和Na(+)-K(+)-ATP酶α1和β1亚基的个体发生情况。生长激素丰度从35 dph起达到恒定的高水平,而IGF-I水平在35 dph时达到峰值,然后显著下降。Na(+)-K(+)-ATP酶α1和β1亚基mRNA在35 dph之前均增加,然而,在46 dph时α1亚基仍保持高水平而β1亚基下降。1 dph幼鱼的Na(+)-K(+)-ATP酶活性较低,但随着发育进程迅速增加。在幼鱼发育的背景下讨论了这些发现的重要性。