Molecular characterization of prothoracicotropic hormone (PTTH) from the giant silkmoth Antheraea pernyi: Developmental appearance of PTTH-Expressing cells and relationship to circadian clock cells in central brain
Molecular characterization of prothoracicotropic hormone (PTTH) from the giant silkmoth Antheraea pernyi: Developmental appearance of PTTH-Expressing cells and relationship to circadian clock cells in central brain
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DOI：
10.1006/dbio.1996.0228
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发表时间：
1996-09-15
影响因子：
2.7
通讯作者：
Reppert, SM
中科院分区：
文献类型：
--
作者：
Sauman, I;Reppert, SM
利用PCR技术克隆了家蚕前胸腺生长激素（PTTH）的cDNA。柞蚕PITH cDNA编码221个氨基酸的前激素原，其在氨基酸水平上分别与家蚕和蓖麻蚕PTTH相同51%和71%。细菌表达的重组A.当注射到去脑的蛹中时，pernyi PTTH会刺激成虫发育。PTTH蛋白（约30 kDa，通过Western印迹）和mRNA（约30 kDa，通过Western印迹）。0.9 kb，通过北方印迹）在脑中表达。免疫细胞化学和原位杂交表明，PTTH蛋白和mRNA共定位于L-NSC III从胚胎发育的第4天到成人的生活，无论是蛋白质或mRNA水平的变化不大，在各种蜕皮。一对表达昼夜节律群蛋白PER的免疫反应性的细胞位于A. pernyi大脑。然而，双标记免疫细胞化学研究表明，PTTH和PER位于不同的细胞。PTTH-和PER-表达细胞之间的紧密解剖位置表明这两个细胞群体之间的通信途径，映射对于PTTH释放的昼夜节律控制是重要的。(C)出版社：Academic Press，Inc.
Using a PCR strategy, we have cloned the cDNA for prothoracicatropic hormone (PTTH) from the giant silkmoth, Antheraea pernyi. The A, pernyi PITH cDNA encodes a preprohormone of 221 amino acids that is 51 and 71% identical at the amino acid level with Bombyx mori and Samia cynthia ricini PTTHs, respectively. Bacterially expressed, recombinant A. pernyi PTTH stimulates adult development when injected into debrained pupae. PTTH protein (ca, 30 kDa by Western blot) and mRNA (ca. 0.9 kb by Northern blot) are expressed in brain. Immunocytochemistry and in situ hybridization show that PTTH protein and mRNA are colocalized in L-NSC III from Day 4 of embryogenesis through adult life, with little variation in either protein or mRNA levels at the various ecdyses. A pair of cells expressing immunoreactivity for the circadian flock protein PER is located in the same region as PTTH-expressing L-NSC III in A. pernyi brain. However, double-label immunocytochemical studies show that PTTH and PER are located in different cells. The close anatomical location between PTTH- and PER-expressing cells suggests routes of communication between these two cell populations that map be important for the circadian control of PTTH release. (C) 1996 Academic Press, Inc.