Zhou Zhong-Hua, Yang Hui-Juan, Chen Ming, Lou Cheng-Fu, Zhang Yao-Zhou, Chen Ke-Ping, Wang Yong, Yu Mei-Lan, Yu Fang, Li Jian-Ying, Zhong Bo-Xiong
College of Animal Sciences, Zhejiang University, Hangzhou 310029, PR China.
J Proteome Res. 2008 Dec;7(12):5103-11. doi: 10.1021/pr800383r.
To gain an insight into the effects of different diets on growth and development of the domesticated silkworm at protein level, we employed comparative proteomic approach to investigate the proteomic differences of midgut, hemolymph, fat body and posterior silk gland of the silkworms reared on fresh mulberry leaves and on artificial diet. Seventy-six differentially expressed proteins were identified by MALDI TOF/TOF MS, and among them, 41 proteins were up-regulated, and 35 proteins were downregulated. Database searches, combined with GO analysis and KEGG pathway analysis revealed that some hemolymph proteins such as Nuecin, Gloverin-like proteins, PGRP, P50 and beta/-N-acetylglucosamidase were related to innate immunity of the silkworm, and some proteins identified in silkworm midgut including Myosin 1 light chain, Tropomyosin 1, Profilin, Serpin-2 and GSH-Px were involved in digestion and nutrition absorption. Moreover, two up-regulated enzymes in fat body of larvae reared on artificial diet were identified as V-ATPase subunit B and Arginine kinase which participate in energy metabolism. Furthermore, 6 down-regulated proteins identified in posterior silk gland of silkworm larvae reared on artificial diet including Ribosomal protein SA, EF-2, EF-1gamma, AspAT, ERp57 and PHB were related to silk synthesis. Our results suggested that the different diets could alter the expression of proteins related to immune system, digestion and absorption of nutrient, energy metabolism and silk synthesis poor nutrition and absorption of nutrition in silkworm. The results also confirmed that the poor nutrient absorption, weakened innate immunity, decreased energy metabolism and reduced silk synthesis are the main reasons for low cocoons yield, inferior filament quality, low survival rate of young larvae and insufficient resistance against specific pathogens in the silkworms fed on artificial diet.
为了从蛋白质水平深入了解不同饲料对家蚕生长发育的影响，我们采用比较蛋白质组学方法，研究了以新鲜桑叶和人工饲料饲养的家蚕中肠、血淋巴、脂肪体和后部丝腺的蛋白质组差异。通过基质辅助激光解吸电离飞行时间串联质谱（MALDI TOF/TOF MS）鉴定出76种差异表达蛋白，其中41种蛋白上调，35种蛋白下调。数据库搜索结合基因本体（GO）分析和京都基因与基因组百科全书（KEGG）通路分析表明，一些血淋巴蛋白如Nuecin、类Gloverin蛋白、肽聚糖识别蛋白（PGRP）、P50和β-N-乙酰氨基葡萄糖苷酶与家蚕的先天免疫有关，家蚕中肠中鉴定出的一些蛋白包括肌球蛋白1轻链、原肌球蛋白1、丝切蛋白、丝氨酸蛋白酶抑制剂-2和谷胱甘肽过氧化物酶（GSH-Px）参与消化和营养吸收。此外，在以人工饲料饲养的幼虫脂肪体中鉴定出的两种上调酶被确定为参与能量代谢的V-ATP酶亚基B和精氨酸激酶。此外，在以人工饲料饲养的家蚕幼虫后部丝腺中鉴定出的6种下调蛋白包括核糖体蛋白SA、延伸因子2（EF-2）、延伸因子1γ（EF-1γ）、天冬氨酸氨基转移酶（AspAT）、内质网蛋白57（ERp57）和热激蛋白70（PHB）与丝蛋白合成有关。我们的结果表明，不同饲料可改变家蚕中与免疫系统、营养消化吸收、能量代谢和丝蛋白合成相关蛋白的表达。结果还证实，营养吸收不良、先天免疫减弱、能量代谢降低和丝蛋白合成减少是导致以人工饲料饲养的家蚕茧产量低、丝质差、幼虫成活率低和对特定病原体抵抗力不足的主要原因。
