饥饿对中间球海胆MYP基因转录表达的影响
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大连海洋大学,大连海洋大学,大连海洋大学,大连海洋大学,大连海洋大学

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国家自然科学基金项目(30800849);辽宁省高校杰出青年学者成长计划(LJQ2011077);大连市科学技术基金(2007J23JH024)


Effect of starvation on expression patterns of the MYP gene in Strongylocentrotus intermedius
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Dalian Ocean University,Dalian Ocean University,,,

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    摘要:

    应用实时定量PCR技术对主要卵黄蛋白(Major yolk protein, MYP)基因在不同饥饿时期中间球海胆的体腔细胞、性腺、肠、胃中的转录表达差异进行了分析。结果表明,在正常状态下,MYP基因在体腔细胞、性腺、肠、胃等不同组织中的转录表达差异明显,肠中的表达量最高,其他组织中的表达量均较低。随饥饿时间的延长,MYP基因在体腔细胞中的表达量先迅速下降,而后稳定在较低水平,实验结束时下降至对照组的1.58%;在性腺中的表达量持续上升,实验结束时上升至对照组的679.75%;在肠中的表达量持续下降,实验结束时下降至对照组的33.33%;在胃中的表达量呈上升趋势,实验结束时上升至对照组的106.52倍。综合来看,饥饿状况下,中间球海胆肠中的MYP表达量持续下降,但仍是MYP的主要合成部位;性腺中MYP表达量持续上升,致使其MYP表达比重上升;胃、体腔细胞中表达量在饥饿过程中虽有变化,但总表达量很少,对MYP的整体表达影响不大。

    Abstract:

    The major yolk protein (MYP) can be found in both genders of sea urchins. Both the immature ovary and testis contain a similar amount of MYP (about 9% of wet mass), constituting about 80% of total protein. This presumably acts as a nutrient reserve during gametogenesis. To determine its main production site and the mechanisms for adapting to starvation in Strongylocentrotus intermedius, MYP mRNA expression levels in intestine, stomach, gonad and coelomocytes were analyzed using real time reverse-transcription polymerase chain reaction. Urchins were starved for 15 days and groups of five individuals were sampled at 5, 10 and 15 days. MYP mRNA was detected in all tissues in nonstarved urchins and the amounts expressed between tissues differed widely. The content was greatest in the intestine, 28.94-fold of that measured in coelomocytes and 39.26-fold of that measured in immature gonads. MYP expression in the stomach-1.83% of that in coelomocytes-was the lowest among the four tissues studied. MYP expression levels decreased sharply after starvation in coelomocytes to 1.29%, 33.33% and 1.58% of the controls (nonstarved) after 5, 10 and 15 days, respectively. MYP mRNA levels increased in the gonads during starvation, to 2.93-, 4.07- and 6.80-fold that of the control group, respectively, but the extent of increase slowed with extended time of fasting. The expression rate during the first 5 days increased by 192.82% but this reduced to 38.98% and 67.01% in the second and third 5-day periods. MYP mRNA expression levels in the stomach increased to 25.81-, 23.43- and 106.52-fold of controls at 5, 10 and 15 days of starvation, respectively. By contrast, levels in the intestine declined continuously during starvation to 67.60%, 50.87% and 33.33% of the control, respectively. Thus, MYP mRNA could be expressed in the intestine, stomach, gonads and coelomocytes in S. intermedius. The intestine was the main site of production, which could be explained from the lack of any large alimentary gland in sea urchins. During starvation, the MYP mRNA expression level decreased in coelomocytes and increased in the stomach, but these levels were still minor compared with the amounts in the intestine and gonads. Interestingly, although MYP synthesis was suppressed in the intestine during starvation, the MYP mRNA expression levels were not declined but increased in gonads during starvation. Thus, the gonads, followed the intestine, provide a second main site of MYP production during starvation. Before the initiation of gametogenesis in sea urchins, the gonads increase in size by accumulating nutrients such as proteins, lipids and carbohydrates in nutritive phagocytes that fill the gonadal lumina. Therefore, it appears that the MYP mRNA expression level does not decline during starvation but increase continuously to provide nutrition for gametogenesis in resisting transient food deprivation.

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秦艳杰,孙博林,李霞,王雪,金迪.饥饿对中间球海胆MYP基因转录表达的影响.生态学报,2012,32(6):1755~1761

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