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首页> 外文期刊>Zoological Science >A circadian neuropeptide, pigment-dispersing Factor-PDF, in the last-summer cicada Meimuna opalifera: cDNA cloning and immunocytochemistry
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A circadian neuropeptide, pigment-dispersing Factor-PDF, in the last-summer cicada Meimuna opalifera: cDNA cloning and immunocytochemistry

机译：上一蝉蝉明尾不透明的昼夜节律神经肽，色素分散因子-PDF：cDNA克隆和免疫细胞化学

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Pigment-dispersing factor (PDF), an 18-amino acid neuropeptide, is a principal circadian neuromodulator functioning downstream of the insect brain's circadian.clock, modulating daily rhythms of locomotor activity. Recently, we found that PDF precursors of the cricket Gryllus bimaculatus comprise a nuclear localization signal (NLS). Moreover, the nuclear localization of PDF immunoreactivity and the translocation of GFP-fused PDF precursor into the nucleus have both been demonstrated. These suggest a fundamental role for PDF peptide in the circadian clock system within the nucleus, in addition to its role in downstream neural events. In the present study, we carried out the cDNA cloning of PDF from adult brains of the last-summer cicada Meimuna opalifera, and found that an isolated clone (545 bp) encodes an ordinary PDF precursor protein. PDF peptide itself shows a high sequence identity (78-94%) and similarity (89-100%) to insect PDFs and also to the crustacean beta-PDH peptides. The computer-assisted sequence analysis of PDF precursor revealed a possible translocation into the nucleus, despite the lack of a definite NLS-like sequence. Using immunocytochemistry, the optic lobes of M. opalifera revealed PDF-immunoreactive neurons in both the medulla and lamina neuropiles. All these PDF cells exhibited prominent immunolabeling of both their perikarya and axons, but not their nuclei. Our results provide the first structural and immunocytochemical identification of PDF neurons in Hemiptera.
机译：色素分散因子（PDF）是一种18个氨基酸的神经肽，是一种主要的昼夜节律神经调节剂，作用于昆虫大脑昼夜节律的下游，调节运动活动的日常节律。最近，我们发现PDF Gryllus bimaculatus的PDF前体包含核定位信号（NLS）。而且，已经证明了PDF免疫反应性的核定位和GFP融合的PDF前体向核内的易位。这些提示了PDF肽除了在下游神经事件中的作用外，在核内生物钟系统中也起着基本作用。在本研究中，我们从最后一个蝉蝉鸣尾成虫的成年脑中进行了PDF的cDNA克隆，发现分离的克隆（545 bp）编码普通的PDF前体蛋白。 PDF肽本身与昆虫PDF以及甲壳类β-PDH肽具有很高的序列同一性（78-94％）和相似性（89-100％）。 PDF前体的计算机辅助序列分析显示，尽管缺乏确定的NLS样序列，但仍可能易位进入细胞核。使用免疫细胞化学技术，不透明支原体的视神经叶揭示了髓质和椎板神经细胞中的PDF免疫反应性神经元。所有这些PDF细胞均显示出其周核细胞和轴突的显着免疫标记，但未显示其细胞核。我们的结果提供了半翅目PDF神经元的第一个结构和免疫细胞化学鉴定。

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