RH 5849, a nonsteroidal ecdysone agonist: effects on a Drosophila cell line

The steroid molting hormone 20-hydroxyecdysone is the physiological inducer of molting and metamorphosis in insects. In ecdysone-sensitive Drosophila Kc cells, the insecticide RH 5849 (1,2-dibenzoyl-1-tert-butylhydrazine) mimics the action of 20-hydroxyecdysone by causing the formation of processes, an inhibition of cell proliferation, and induction of acetylcholinesterase. RH 5849 also competes with [3H]ponasterone A for high-affinity ecdysone receptor sites from Kc cell extracts. Resistant cell populations selected by growth in the continued presence of either RH 5849 or 20-hydroxyecdysone are insensitive to both compounds and exhibit a decreased titer of measurable ecdysone receptors. Although it is less potent than 20-hydroxyecdysone in both whole-cell and cell-free receptor assays, RH 5849 is the first nonsteroidal ecdysone agonist.     

Insect hormones: alpha ecdysone and 20-hydroxyecdysone in bracken fern

The two major molting hormones of insects, alpha ecdysone and 20-hydroxyecdysone, were isolated in crystalline form from dry pinnae of the bracken fern, Pteridium aquilinum (L.) Kuhn. Three unidentified substances with molting hormone activity were also detected. Bracken is the first plant found to contain both of the major insect ecdysones, and it is the first known plant source of alpha ecdysone.     

Ecdysones and analogs: effects on development and reproduction of insects

Ingestion of certain synthetic ecdysone analogs inhibited larval growth and development in several species of insects, whereas 20-hydroxyecdysone was inactive or considerably less active. Natural 20-hydroxyecdysone and ponasterone A, and a synthetic ecdysone analog inhibited ovarian maturation and egg production in the adult housefly. These effects appeared to be related to hormonal activity.     

26-hydroxyecdysone: new insect molting hormone from the egg of the tobacco hornworm

Five kilograms of tobacco hornworm eggs (48 to 64 hours old) afforded 26.5 milligrams of a new crystalline insect molting hormone identified as 26-hydroxyecdysone. The three known insect ecdysones-alpha-ecdysone, 20-hydroxyecdysone, and 20,26-dihydroxyecdysone-were also present but in much smaller quantities. The new hormone is the predominant molting hormone in the hornworm during this stage of embryonic development. These results takent in context with the current knowledge of the chemistry and biochemistry of the molting hormones during postembryonic development in the hornworm indicate quantitative and qualitative differences in the biosynthetic-mnetabolic pathways as well as in the ecdysones in different developmental stages of this insect.     

Ecdysone analog: conversion to alpha ecdysone and 20-hydroxyecdysone by an insect

The tritium-labeled synthetic ecdysone analog Delta(7)-5beta-cholestene-2beta,-3beta,14alpha-triol-6-one terminated diapause when injected into diapausing tobacco hornworm pupae and was converted into tritium-labeled alpha-ecdysone and tritium-labeled 20-hydroxyecdysone. About half of the crystalline alpha-ecdysone and 20-hydroxyecdysone isolated from the tobacco hornworms 7(1/2) to 8(1/2) days after injection was derived from endogenous steroid precursors and half from the transformation of the synthetic ecdysone analog.     

Peptide and steroid regulation of muscle degeneration in an insect

Two types of cell death occur in the intersegmental muscles of the giant silkmoth Antheraea polyphemus. The first results from a slow atrophy of the fibers, and the second is a rapid, programmed dissolution of the muscle. Both types appear to be mediated by endocrine factors. The slow atrophy is brought about by the decline in the steroid molting hormone 20-hydroxyecdysone and can be prevented with exogenous steroid. The rapid degeneration is triggered by the peptide eclosion hormone, but the sensitivity of the muscle to the peptide depends on the history of exposure of the muscle to 20-hydroxyecdysone.     

中华绒螯蟹在“蟹龙宫”饲养环境中的个体蜕壳与生长

分析"蟹龙宫"饲养中华绒螯蟹(Eriocheir sinensis)在一个蜕壳周期的蜕壳与生长情况。结果发现:"蟹龙宫"饲养中华绒螯蟹蜕壳后体质量平均增长率为33.19%,头胸甲长、头胸甲宽和体高的平均增长率为12.12%～14.97%,平均蜕壳间期为59.35 d。蜕壳前平均肥满度达60.84%,头胸甲的色度值(RGB)平均为109.17,蜕皮激素(EH)含量达14.18 IU/mL。试验河蟹在蜕壳前生长性状、EH含量和蜕壳间期长短与蜕壳后的增长率不存在显著相关性(P0.05)。在"蟹龙宫"的个体养殖系统中,中华绒螯蟹的肥满度达60%,蜕皮激素含量升至14 IU/mL时会启动蜕壳。     

藤壶金星幼虫附着变态机制

藤壶属节肢动物门(Arthropoda)甲壳亚门(Crustacea)蔓足下纲(Cirripedia)围胸总目(Thoracica), 具备特殊的形态结构、生活史和种群生态特征,是最主要的海洋污损生物。其幼虫阶段通常经历6期无节幼体和1期不摄食的金星幼虫,从浮游的金星幼虫附着变态成固着的稚体是藤壶生活史中的一个关键环节。外界化学和生物因子中成体提取物、水溶性信息素、足迹、神经递质、激素、生物膜等均影响藤壶金星幼虫的附着变态;内在因子即金星幼虫的生理状态(能量储量和年龄)决定了其对外界因子的反应程度。概括了近年来藤壶附着变态生理机制和分子机制研究的进展,可为深入了解藤壶金星幼虫附着变态机制提供参考,也为开发新型、高效、环保的防污剂提供理论指导。     

抑食肼对大型溞的急慢性毒性效应研究

选择21%抑食肼可湿性粉剂为受试农药,研究其对大型溞(Daphnia magna)的急性毒性和21 d慢性毒性效应,观察大型溞各个繁殖和生长指标变化,建立毒物剂量-反应关系,并寻找其中的敏感生物学指标。急性毒性试验结果表明,21%抑食肼可湿性粉剂对大型溞的48 h-EC50为0.94 mg·L-1;慢性毒性试验结果表明,抑食肼对大型溞的初次蜕壳时间、蜕壳次数、产卵时间、产幼溞数和产幼溞胎数等都有显著的影响,其中,0.135 mg·L-1的暴露浓度可以显著减少大型溞蜕壳次数、产幼溞总数量和产幼溞胎数,综合各个繁殖毒性终点指标,21 d无可见效应浓度(NOEC)为0.045 mg·L-1。此外,研究还发现低至0.015 mg·L-1的抑食肼暴露21 d可以使母溞体长显著性下降。研究结果表明,抑食肼胁迫在一定程度上对大型溞的生长和繁殖具有抑制效应。     

Biosynthesis of agr-Ecdysone by Prothoracic Glands in vitro

An in vitro study in which isolated prothoracic glands of the Bombyx silkworm were cultured has provided definite evidence that the prothoracic gland is the site where molting hormone is synthesized. The hormone behaved very similarly to free ecdysone on thin-layer chromatography. Analysis by liquid chromatography and mass fragmentography revealed that the hormone is identical with alpha-ecdysone.     

Bracken and locust ecdysones: their effects on molting in the desert locust

Bracken contains ecdysone derivatives that are active when injected into locusts. However, when fed to the desert locust as its sole or chief diet, it does not affect molting, growth, or development. There is evidence that, in locusts, the active ecdysones are dehydroxylated to (alpha)-ecdysone and passed out through the gut in the feces. There is no evidence for any uptake of ecdysones from the gut.     

Essential roles for ecdysone signaling during Drosophila mid-embryonic development

Although functions for the steroid hormone ecdysone during Drosophila metamorphosis have been well established, roles for the embryonic ecdysone pulse remain poorly understood. We show that the EcR-USP ecdysone receptor is first activated in the extraembryonic amnioserosa, implicating this tissue as a source of active ecdysteroids in the early embryo. Ecdysone signaling is required for germ band retraction and head involution, morphogenetic movements that shape the first instar larva. This mechanism for coordinating morphogenesis during Drosophila embryonic development parallels the role of ecdysone during metamorphosis. It also provides an intriguing parallel with the role of mammalian extraembryonic tissues as a critical source of steroid hormones during embryonic development.     

家蚕β-呋喃果糖苷酶基因BmSuc1表达特征及其对激素的响应

【目的】探究BmSuc1基因在家蚕不同组织及不同时期的表达特征及经昆虫激素处理后的表达变化规律,明确昆虫激素对BmSuc1基因的调控作用,为深入解析BmSuc1基因的功能及其表达调控机制提供参考依据。【方法】利用实时荧光定量PCR检测BmSuc1基因在家蚕发育过程中不同时期和不同组织及外源激素处理后的表达特征,并通过双链RNA （dsRNA）干扰试验检测家蚕20-羟基蜕皮素（20E）受体基因（USP）干扰效果及BmSuc1基因的表达变化。【结果】BmSuc1基因在家蚕中肠中的相对表达量最高,其次是丝腺、表皮和血淋巴,在其他组织中的相对表达量很低或几乎不表达;BmSuc1基因呈脉冲型的表达模式,在家蚕每个龄期的将眠时、五龄后期（上簇前）及预蛹时的相对表达量较高。利用外源激素[20E和保幼激素（JH）]分别处理五龄第2 d发育良好的家蚕,发现经20E处理后12和18 h,BmSuc1基因相对表达量极显著高于注射0.1%二甲基亚砜（DMSO）的对照组（P<0.01,下同）,但在测定时间范围内JH处理组的BmSuc1基因相对表达量与对照组间无显著差异（P>0.05）。以体外转录合成USP基因的dsRNA注射五龄第3 d家蚕,注射后24和36 h,USP基因相对表达量极显著下调,即dsRNA干扰成功。利用RNAi技术干扰USP基因能阻断20E信号转导,致使BmSuc1基因表达受抑制,其相对表达量在干扰USP基因后24和36 h显著下调（P<0.05）。【结论】BmSuc1基因主要在家蚕蜕皮和变态时高表达,暗示其可能参与家蚕的变态发育过程。添加外源20E可诱导BmSuc1基因转录,而阻断20E信号转导途径会抑制BmSuc1基因表达,即BmSuc1基因作为20E信号转导通路中的下游靶基因,直接或间接受到20E信号调控。     

苯甲酰基脲类和肼类昆虫生长调节剂对棉铃虫的生物活性

研究了作用机制完全不同的苯甲酰基脲类（氟铃脲）和双苯甲酰基肼类（抑食肼和虫酰肼）昆虫生长调节剂（ＩＧＲ）及其混用对棉铃虫的生物活性和作用方式。供试３龄幼虫经氟铃脲点滴处理后,逐渐表现活动缓慢、反应迟钝、取食减少,最终死亡。死亡幼虫体壁薄而易破,头部背面呈现明显突起的白色透明水泡状（易破体并有体液流出）,部分虫体黑化,表现明显的蜕皮障碍。３龄幼虫经抑食肼处理后,最先也表现为行为迟缓、食量减小,最终死亡的幼虫虫体缩小并发黑,部分存活的幼虫２ｄ后即蜕皮。氟铃脲与抑食肼混合处理后,部分试虫呈现氟铃脲的中毒症状,部分试虫则表现抑食肼中毒症状,少量个体两种症状并发。氟铃脲对棉铃虫３幼虫的ＬＤ５０值（点滴法）和１龄幼虫的ＬＣ５０值（叶片法）分别为０．７４３４μｇａ．ｉ．／头和２．０５９２ｍｇ／Ｌ,表现出很强的触杀作用和胃毒作     

Biosynthesis of agr and beta-Ecdysones from Cholesterol outside the Prothoracic Gland in Bombyx mori

Labeling experiments have established that cholesterol is converted into alpha-and beta-ecdysones in isolated abdomens of silkworm larvae. Since the isolated abdomens do not contain the prothoracic glands, a doubt is cast on the long-standing principle in insect endocrinology that the prothoracic glands are the source of ecdysone secretion.     

马铃薯甲虫LdATPaseE调控幼虫蜕皮的分子机制

[目的]研究LdATPaseE通过类胰岛素信号通路影响保幼激素和蜕皮激素通路,调节马铃薯甲虫幼虫化蛹的分子机制.[方法]LdATPaseE cDNA序列通过马铃薯甲虫转录组数据分析和RT-PCR克隆获得;qPCR检测其在马铃薯甲虫各发育阶段,以及四龄幼虫不同组织的相对表达量;采用喂食幼虫dsRNA的方法,分析该基因在马...     

苹果蠹蛾几丁质脱乙酰基酶2的基因克隆、表达模式和分子特性

【目的】研究苹果蠹蛾（Cydia pomonella）几丁质脱乙酰基酶2（chitin deacetylase 2,CDA2）的分子特性和表达模式,为新型杀虫剂靶标筛选提供理论依据。【方法】基于苹果蠹蛾转录组和基因组数据,通过同源比对获得苹果蠹蛾CDA2的相关信息;新鉴定的CpCDA2a和CpCDA2b与其他昆虫中已鉴定的CDA2氨基酸序列进行多序列比对,并采用MEGA7软件邻接法（neighbor-joining,NJ）构建系统发育树;通过生物信息软件预测CpCDA2a和CpCDA2b两个剪切体的蛋白功能域和三维结构并分析其差异,并通过在线软件对其分子量、等电点和亲/疏水性等蛋白理化性质及糖基化修饰位点等方面进行分析比较;采用荧光定量PCR检测CpCDA2a和CpCDA2b两个剪切体在苹果蠹蛾不同发育时期及其幼虫不同组织的表达情况,以及注射蜕皮激素后其表达量的变化趋势。【结果】获得了苹果蠹蛾CDA2的两个可变剪切体CpCDA2a和CpCDA2b的全长CDS序列及其内含子和外显子位置信息;分析发现两个剪切体的蛋白序列均具有信号肽、几丁质结合域（ChBD）、低密度脂蛋白受体域（LDLa）和几丁质脱乙酰基催化域（CDA）等功能域。多序列比对和聚类分析结果表明,不同昆虫间CDA2差异剪切类型非常类似,相同目昆虫的序列以较高的置信度聚为一支。三维结构模拟与比较发现两个剪切体的ChBD功能域在结构上存在较大差异;理化性质分析显示两个剪切体的亲/疏水性和糖基化位点也存在差异。不同发育时期表达量分析表明,CpCDA2a主要在幼虫期高表达,CpCDA2b主要在蛹的前期和中期高表达,而且在幼虫蜕皮前和蜕皮后CpCDA2a和CpCDA2b表达量均会显著上调;不同组织的表达量分析表明,CpCDA2a和CpCDA2b均在表皮中表达量最高,而CpCDA2b在头部的表达量也较高。对注射蜕皮激素后幼虫中CpCDA2a和CpCDA2b表达量分析显示,与对照相比在注射蜕皮激素后CpCDA2a的表达量在24 h和48 h均有所上调,但差异不显著,而CpCDA2b在24 h和48 h均显著上调。【结论】苹果蠹蛾CDA2的转录本存在两个可变剪切体CpCDA2a和CpCDA2b,根据对其分子特性、发育阶段和组织表达谱、以及注射蜕皮激素后表达动态等方面的综合分析,推测两者均参与苹果蠹蛾蜕皮和新表皮形成过程,但由于可变剪切的序列差异导致的蛋白结构和理化性质上的差异造成了两者在功能上的分化。     

Induction of follicle separation in the mosquito by physiological amounts of ecdysterone

Physiological quantities of the molting hormone, ecdysterone, injected into female Aedes aegypti prematurely induced separation of incipient follicles in the ovarioles, an event that normally occurs only in blood-fed females. It was possible to stimulate this morphological event with physiological amounts of hormone by mimicking, with two injections, the timing of normal increases in endogenous hormone of blood-fed females.