蛋白质组学分析揭示了卡尼鄂拉蜜蜂下咽腺活动的蛋白质表达差异。
Proteomics analysis reveals protein expression differences for hypopharyngeal gland activity in the honeybee, Apis mellifera carnica Pollmann.
作者信息
Ji Ting, Liu Zhenguo, Shen Jie, Shen Fang, Liang Qin, Wu Liming, Chen Guohong, Corona Miguel
机构信息
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, China.
出版信息
BMC Genomics. 2014 Aug 8;15(1):665. doi: 10.1186/1471-2164-15-665.
BACKGROUND
Most of the proteins contained in royal jelly (RJ) are secreted from the hypopharyngeal glands (HG) of young bees. Although generic protein composition of RJ has been investigated, little is known about how age-dependent changes on HG secretion affect RJ composition and their biological consequences. In this study, we identified differentially expressed proteins (DEPs) during HG development by using the isobaric tag for relative and absolute quantification (iTRAQ) labeling technique. This proteomic method increases the potential for new protein discovery by improving the identification of low quantity proteins.
RESULTS
A total of 1282 proteins were identified from five age groups of worker bees, 284 of which were differentially expressed. 43 (15.1%) of the DEPs were identified for the first time. Comparison of samples at day 6, 9, 12, and 16 of development relative to day 3 led to the unambiguous identification of 112, 117, 127, and 127 DEPs, respectively. The majority of these DEPs were up-regulated in the older worker groups, indicating a substantial change in the pattern of proteins expressed after 3 days. DEPs were identified among all the age groups, suggesting that changes in protein expression during HG ontogeny are concomitant with different states of worker development. A total of 649 proteins were mapped to canonical signaling pathways found in the Kyoto Encyclopedia of Genes and Genomes (KEGG), which were preferentially associated with metabolism and biosynthesis of secondary metabolites. More than 10 key high-abundance proteins were involved in signaling pathways related to ribosome function and protein processing in the endoplasmic reticulum. The results were validated by qPCR.
CONCLUSION
Our approach demonstrates that HG experienced important changes in protein expression during its ontogenic development, which supports the secretion of proteins involved in diverse functions in adult workers beyond its traditional role in royal jelly production.
背景
蜂王浆（RJ）中含有的大多数蛋白质是由幼蜂的下咽腺（HG）分泌的。尽管已经对RJ的一般蛋白质组成进行了研究，但关于HG分泌随年龄的变化如何影响RJ组成及其生物学后果，人们知之甚少。在本研究中，我们使用相对和绝对定量的等压标签（iTRAQ）标记技术，鉴定了HG发育过程中差异表达的蛋白质（DEP）。这种蛋白质组学方法通过改进低丰度蛋白质的鉴定，增加了发现新蛋白质的潜力。
结果
从五个年龄组的工蜂中总共鉴定出1282种蛋白质，其中284种差异表达。43种（15.1%）DEP是首次鉴定出来的。将发育第6、9、12和16天的样本与第3天的样本进行比较，分别明确鉴定出112、117、127和127种DEP。这些DEP中的大多数在较老的工蜂组中上调，表明3天后蛋白质表达模式发生了实质性变化。在所有年龄组中都鉴定出了DEP，这表明HG个体发育过程中蛋白质表达的变化与工蜂发育的不同状态相伴。共有649种蛋白质被映射到京都基因与基因组百科全书（KEGG）中发现的经典信号通路，这些通路优先与次生代谢物的代谢和生物合成相关。超过10种关键的高丰度蛋白质参与了与核糖体功能和内质网中蛋白质加工相关的信号通路。结果通过qPCR得到验证。
结论
我们的方法表明，HG在其个体发育过程中经历了重要的蛋白质表达变化，这支持了参与成年工蜂多种功能的蛋白质的分泌，而不仅仅是其在蜂王浆生产中的传统作用。
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