College of Animal Science and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People's Republic of China.
Mol Biol Rep. 2012 Mar;39(3):2665-75. doi: 10.1007/s11033-011-1020-y. Epub 2011 Jun 22.
Lipid storage droplet 1 (LSD-1), a PAT family protein located around lipid droplets in insects, is intimately linked to lipid droplets formation and lipid metabolism. Conjugated linoleic acid (CLA) and rosiglitazone (Rosi) have previously been shown to modulate the expression of several PAT family proteins through peroxisome proliferator-activated receptor-γ (PPARγ). In the present study, we isolated and characterized a novel LSD-1 gene, referred to AccLSD-1, from Chinese honeybee (Apis cerana cerana). Sequence analysis indicated that the central region of LSD-1 protein had significant sequence similarity and a typical LSD-1 gene was composed of 8 exons and 7 introns. Interestingly, the first intron of AccLSD-1 including several PPARγ-response elements (PPREs) was located in 5' UTR. Analysis of 5'-flanking region of AccLSD-1 revealed a number of putative cis-acting elements, including three PPREs. Quantitative real-time PCR showed that AccLSD-1 expressed ubiquitously from feeding larva to adult, and its expression level was highest at brown-eyed pupae (Pb) stage. The effect of CLA, Rosi and combination on AccLSD-1 expressions indicated 1% CLA and 0.5 mg/ml Rosi were considered as the suitable diets for rearing adult workers in laboratory, and AccLSD-1 was down-regulated by CLA whereas up-regulated by Rosi. Furthermore, the combination of CLA and Rosi remarkly rescued the suppression of AccLSD-1 expression by CLA alone. These results suggest that AccLSD-1 is associated with A. cerana cerana development, especially during pupal metamorphosis, and can be regulated by CLA or Rosi possibly via activating PPARγ.
脂滴 1（LSD-1）是昆虫中位于脂滴周围的 PAT 家族蛋白，与脂滴形成和脂质代谢密切相关。共轭亚油酸（CLA）和罗格列酮（Rosi）先前已被证明通过过氧化物酶体增殖物激活受体-γ（PPARγ）调节几种 PAT 家族蛋白的表达。在本研究中，我们从中华蜜蜂（Apis cerana cerana）中分离并鉴定了一种新的 LSD-1 基因，称为 AccLSD-1。序列分析表明，LSD-1 蛋白的中心区域具有显著的序列相似性，典型的 LSD-1 基因由 8 个外显子和 7 个内含子组成。有趣的是，AccLSD-1 的第一个内含子包括几个 PPARγ 反应元件（PPREs），位于 5'UTR 中。AccLSD-1 5'-侧翼区分析显示了许多潜在的顺式作用元件，包括三个 PPREs。定量实时 PCR 显示 AccLSD-1 从取食幼虫到成虫广泛表达，其表达水平在棕眼蛹（Pb）期最高。CLA、Rosi 及其组合对 AccLSD-1 表达的影响表明，1%CLA 和 0.5mg/ml Rosi 被认为是实验室饲养成年工蜂的适宜饮食，CLA 下调 AccLSD-1 表达，而 Rosi 上调 AccLSD-1 表达。此外，CLA 和 Rosi 的组合显著挽救了 CLA 单独对 AccLSD-1 表达的抑制作用。这些结果表明，AccLSD-1 与 A. cerana cerana 的发育有关，特别是在蛹变态期间，并且可以通过激活 PPARγ 被 CLA 或 Rosi 调节。
