昆虫促前胸腺激素研究进展

昆虫促前胸腺激素研究进展李毅平龚和（中国科学院动物研究所,北京１０００８０）关键词促前胸腺激素受体信号系统促前胸腺激素（ｐｒｏｔｈｏｒａｃｉｃｏｔｒｏｐｉｃｈｏｒ－ｍｏｎｅＰＴＴＨ）因其促进前胸腺（ＰＧ）合成和分泌蜕皮激素而得名,以前也称为脑激素,因...     

家蚕FXPRL神经肽前体基因的体外转录

昆虫滞育激素（Ｄｉａｐａｕｓｅｈｏｒｍｏｎｅ：ＤＨ）、性信息素合成激活肽（Ｐｈｅｒｏｍｏｎｅｂｉｏｓｙｎｔｈｅｓｉｓａｃｔｉｖａｔｉｎｇｎｅｕｒｏｐｅｐｔｉｄｅ：ＰＢＡＮ）是诱导昆虫滞育和性信息素（Ｓｅｘｐｈｅｒｏｍｏｎｅ）合成的两个重要神经肽［１...     

植源性昆虫生长调节剂

昆虫中存在一类含量少而生理作用极为显著的活性物质,其中包括昆虫激素、昆虫信息素和昆虫毒素等。昆虫激素又称变态激素,一般包括脑激素、保幼激素和蜕皮激素等。昆虫的胚胎后发育与成熟过程是受体内这些激素控制的。若破坏激素系统的平衡,如外加过量的这种激素或对抗这种激素的物质,就会阻碍昆虫的正常生长、分化和变态的程     

保幼激素及其代谢产物的HPLC分离方法的改进和应用

传统的正相高效液相色谱（normal phase high performance liquid chromatography, NP-HPLC）可以较好地分离保幼激素（juvenile hormone, JH）Ⅰ、Ⅱ和Ⅲ,但不能分离保幼激素的代谢产物及其类似物。经过改进的反相高效液相色谱（reverse phase high performance liquid chromatography, RP-HPLC）不仅可以很好地分离保幼激素,还能定性定量分析保幼激素的代谢产物及其类似物。离体培养昆虫咽侧体（corpora allata, CA）所合成的、被同位素标记的痕量保幼激素可以用以上两种色谱方法进行分离和鉴定。此外,RP-HPLC还可以用来分离体内或体外同位素标记的保幼激素代谢产物,以及测量血淋巴中的保幼激素滴度。     

蜱类的激素

激素在节肢动物的发育和生殖中起重要作用。它调节昆虫的生长、蜕皮、变态、生殖和其他许多生理过程。然而,蝉类激素的研究远远落后于昆虫,到目前为止,还没有明确确定蝉类具内分泌功能的腺体,并且保幼激素的存在只是推测的。自Delbecque[1]首次在蝉类中发现锐皮激素以来,国外已做了部分工作,国内也开始起步。20世纪中有3本出色的著作介绍蝉类内分泌和激素的内容与进展。分别是Solomon著的“PhysiologyofTicks”（1982年版）,Saner和Hair著的“Morphology,PhysiologyandBehaviouralBiologyofTicks”（1986年版）和Sonenstune著…     

家蚕滞育激素－性信息素合成激活肽基因的表达

家蚕滞育激素－性信息素合成激活肽基因的表达徐卫华（中国农业科学院蚕业研究所,江苏镇江,２１２０００）山下兴亚（名古屋大学农学院,日本名古屋,４６４－０１）关键词滞育激素－性信息素合成激活肽基因;发育阶段;表达;家蚕昆虫是地球上最繁盛的物种,占地球上生...     

半翅目昆虫卵黄原蛋白及其合成调控的研究进展

昆虫卵黄原蛋白（Vitellogenins, Vg）是一种多功能的生殖发育关键调控蛋白,在不同昆虫体内的结构、合成调控及功能不尽相同。随着基因编辑技术的成熟,运用分子手段调控Vg的合成,可减少卵黄发生,降低昆虫的繁殖力,成为有效防治害虫的优势方法之一。因此,Vg及其合成调控的研究受到广泛关注。半翅目害虫是农林业的重点防治对象之一,除直接刺吸为害寄主外,其常传播植物病原体,对农业生产造成了严重危害。半翅目昆虫Vg除在生殖发育中的关键作用外,还与病原菌的传播、寄主免疫等密切相关,可成为分子水平防治半翅目害虫及其继发病害的优势靶标。因此,本文总结了半翅目昆虫Vg的合成方式、合成场所,指明了其结构上蛋白亚基数目的差异,概述了其与昆虫免疫反应、植物防御、病毒传播等有关的研究进展,总结了其合成的保幼激素（包括保幼激素受体Methoprene-tolerant和转录因子Krüppel homolog 1等关键调控因子等）、蜕皮激素和胰岛素信号通路等主要的内分泌激素调控通路,以及以营养信号调控为主的非激素调控通路,为探索半翅目害虫的分子防控手段提供理论依据。     

无脊椎动物内分泌与激素综览

本文简要介绍了近年对非昆虫无脊椎动物内分泌与激素研究动态和各大门类内分泌与激素较成熟的认识。现已明确相似于脊椎动物的内分泌也存在于许多无脊椎动物且有不同功能的多种激素。神经内分泌细胞及其分泌的神经激素、蜕皮激素等激素了解更为系统、深入，研究最多的动物是节肢动物（特别是甲壳类）。     

小菜蛾鞣化激素基因克隆及其功能分析

鞣化激素是调节昆虫表皮骨化和翅膀发育的一种神经激素, 尽管已经在许多不同种昆虫上克隆了鞣化激素基因, 但是关于小菜蛾 Plutella xylostella鞣化激素及其基因的研究至今未见报道。本研究克隆了两个小菜蛾鞣化激素基因Pxbursα和Pxbursβ （GenBank 登录号分别为KF498645和KF498646）全长cDNA, 其序列长度分别为537 bp和360 bp, 与已报道的其他昆虫的鞣化激素氨基酸序列一致性分别为51%~68% 和37%~57%。实时定量PCR分析发现Pxbursα和Pxbursβ均在蛹期表达量高, 而在幼虫期和成虫期的表达量低。以Pxbursα部分序列的双链RNA（dsRNA）饲喂小菜蛾4龄末期幼虫, 发现蛹期Pxbursα的表达受到了显著抑制, 小菜蛾的发育停滞在蛹期而无法正常羽化, 并最终死亡。由此推测, 小菜蛾鞣化激素基因在蛹期的大量表达对其生长发育和羽化具有重要的作用。     

昆虫鞣化激素研究进展

鞣化激素是调控昆虫体壁黑化及翅伸展的一类激素, 是由BURS和PBURS两个亚基组成的一种异源二聚体蛋白质。BURS和PBURS亚基在结构及其进化上相对较为保守, 氨基酸序列中均含有11个半胱氨酸残基。鞣化激素主要是在胸腹神经节中合成的, 一旦释放到血淋巴就与其受体LGR2结合进而激活cAMP/PKA信号, 从而促进酪氨酸羟化酶(tyrosine hydroxylase, TH)的磷酸化。活化后的TH将酪氨酸(tyrosine)转变为多巴（DOPA）, 引起昆虫表皮鞣化。同时, cAMP/PKA信号也引起翅真皮细胞凋亡从而促进翅的伸展。除了鞣化激素异聚体调控表皮鞣化及翅的伸展外, BURS亚基或PBURS亚基组成的同源二聚体经IMD路径, 激活转录因子Relish调控昆虫的免疫反应。本文就鞣化激素分子结构特性、 作用机制及功能等方面的研究进展进行了综述, 旨在为进一步研究昆虫鞣化激素提供借鉴和参考。     

昆虫神经肽allatostati与allatotropin的研究新近展

Recently, two families of insect neuropeptides, the allatostatin, andallatotropin, have been identified. All allatostatins and allatotropins identified so farare neurosecretory polypeptides. A 13-amino acid allatotropin has been identifiedfrom adult Manduca sexta. A group of five structurally related allatostatins has beenidentified from Diploptara punctata. They either inhibit (allatostatin) or stimulate(allatotropin) the production of JH by CA. In this paper recent advances in researchon insect neuropeptides AS and AT and their biological significance are reported.     

Triple co-localisation of two types of allatostatin and an allatotropin in the frontal ganglion of the lepidopteran Lacanobia oleracea (Noctuidae): innervation and action on the foregut

The triple co-localisation of peptidergic material immunoreactive to antisera raised against allatostatins of the Y/FXFGL-NH2 type, Manduca sexta allatostatin (Mas-AS), and allatotropin has been demonstrated in a single pair of anterodorsal neurones in the frontal ganglion of the tomato moth, Lacanobia oleracea (Noctuidae). Another pair of posterior neurones contain only Y/FXFGL-NH2-type allatostatin immunoreactivity. The neurites of all four cells trifurcate, and axons project to the brain in the frontal connectives and to the foregut in the recurrent nerve. Axons from the anterior neurones, within the recurrent nerve, have prominent lateral branches supplying muscles of the crop, and axons from both anterior and posterior cells show profuse branching and terminal arborisations in the region of the stomodeal valve. The brain contributes Y/FXFGL-NH2-immunoreactive material, but not allatotropin or Mas-AS, to the recurrent nerve via NCC 1+2 and NCC 3. All three peptides have a reversible effect on the spontaneous (peristaltic) contractions of the foregut (crop) in vitro. Thus, both types of allatostatin are inhibitory at 10(-12) to 10(-7) M, whereas allatotropin is strongly myostimulatory at 10(-14) M. This is the first demonstration of the gut myoinhibitory effects of Mas-AS and, taken together with the effects of Y/FXFGL-NH2-type allatostatins and allatotropin, reveals a different functional aspect to that normally attributed to these three peptides, i.e. control of juvenile hormone synthesis by the corpus allatum.     

Molecular cloning and genomic organization of an allatostatin preprohormone from Drosophila melanogaster

The insect allatostatins are neurohormones, acting on the corpora allata (where they block the release of juvenile hormone) and on the insect gut (where they block smooth muscle contraction). We screened the "Drosophila Genome Project" database with electronic sequences corresponding to various insect allatostatins. This resulted in alignment with a DNA sequence coding for some Drosophila allatostatins (drostatins). Using PCR with oligonucleotide primers directed against the presumed exons of this Drosophila allatostatin gene and subsequent 3'- and 5'-RACE, we were able to clone its cDNA. The Drosophila allatostatin preprohormone contains four amino acid sequences that after processing would give rise to four Drosophila allatostatins: Val-Glu-Arg-Tyr-Ala-Phe-Gly-Leu-NH(2) (drostatin-1), Leu-Pro-Val-Tyr-Asn-Phe-Gly-Leu-NH(2) (drostatin-2), Ser-Arg-Pro-Tyr-Ser-Phe-Gly-Leu-NH(2) (drostatin-3), and Thr-Thr-Arg-Pro-Gln-Pro-Phe-Asn-Phe-Gly-Leu-NH(2) (drostatin-4). Drostatin-2 is identical to helicostatin-2 (11-18) and drostatin-3 to helicostatin-3, two neurohormones previously isolated from the moth Helicoverpa armigera. Furthermore, drostatin-3 has previously been isolated from Drosophila itself. Drostatins-1 and -4 are novel members of the insect allatostatin neuropeptide family. The Drosophila allatostatin preprohormone gene contains two introns and three exons. The gene is located on the right arm of the third chromosome, position 96A-B. The existence of at least four different Drosophila allatostatins opens the possibility of a differential action of some of these hormones on the two recently cloned Drosophila allatostatin receptors, DAR-1 and -2. This is the first report on an allatostatin preprohormone from Drosophila.     

Allatoregulatory peptides in Lepidoptera, structures, distribution and functions

Allatoregulatory peptides either inhibit (allatostatins) or stimulate (allatotropins) juvenile hormone (JH) synthesis by the corpora allata (CA) of insects. However, these peptides are pleitropic, the regulation of JH biosynthesis is not their only function. There are currently three allatostatin families (A-, B-, and C-type allatostatins) that inhibit JH biosynthesis, and two structurally unrelated allatotropins. The C-type allatostatin, characterised by its blocked N-terminus and a disulphide bridge between its two cysteine residues, was originally isolated from Manduca sexta. This peptide exists only in a single from in Lepidoptera and is the only peptide that has been shown to inhibit JH synthesis by the CA in vitro in this group of insects. The C-type allatostatin also inhibits spontaneous contractions of the foregut. The A-type allatostatins, which exist in multiple forms in a single insect, have also been characterised from Lepidoptera. This family of peptides does not appear to have any regulatory effect on JH biosynthesis, but does inhibit foregut muscle contractions. Two structurally unrelated allatotropins stimulate JH biosynthesis in Lepidoptera. The first was identified in M. sexta (Manse-AT) and occurs in other moths. The second (Spofr AT2) has only been identified in Spodoptera frugiperda. Manduca sexta allatotropin also stimulates heart muscle contractions and gut peristalsis, and inhibits ion transport across the midgut of larval M. sexta. The C-terminal (amide) pentapeptide of Manse-AT is important for JH biosynthesis activity. The most active conformation of Manse-AS requires the disulphide bridge, although the aromatic residues also have a significant effect on biological activity. Both A- and C-type allatostatins and Manse-AT are localised in neurosecretory cells of the brain and are present in the corpora cardiaca, CA and ventral nerve cord, although variations in localisation exist in different moths and at different stages of development. The presence of Manse-AS and Manse-AT in the CA correlates with the biological activity of these peptides on JH biosynthesis. There is currently no explanation for the presence of A-type allatostatins in the CA. The three peptide types are also co-localised in neurosecretory cells of the frontal ganglion, and are present in the recurrent nerve that supplies the muscles of the gut, particularly the crop and stomodeal valve, in agreement with their role in the regulation of gut peristalsis. There is also evidence that they are expressed in the midgut and reproductive tissues.     

Neuropepride Regulators of Insect Corpora Allata

SYNOPSIS. Neuropeptides of the insect brain that regulate juvenilehormone synthesis by the corpora allata include allatotropins,stimulatory modulators, and allatostatins, inhibitory modulators.A radiochemical assay for juvenile hormone synthesis by corporaallata in vitro was utilized in the high pressure liquid chromatographicisolation of brain neuropeptides leading to the determinationof their primary structure. Identified are an allatotropin andan allatostatin from a Lepidopteran, Manduca sexta, and a familyof five allatostatins from a Dictyopteran, Diploptera punctata.These neuropeptides are all unique, effective at low concentration(10^–10 to 10^–8 M), act quickly (within hrs) andappear to be effective only within the same order of insectsas that from which the peptides were isolated. The physiologicalstate of the corpora allata conditions the effectiveness ofthe allatostatins of D. punctata. These neuropeptide regulatorsof corpora allatal function may have multiple regulatory roles.This is indicated for D. punctata allatostatin I by specificreceptors in brain and fat body as well as in corpora allatalmembrane preparations, and also by immunocytochemical localizationof allatostatin I in medial nerve cells that terminate withinthe brain as well as in the lateral neurosecretory cells thatterminate on corpus allatum cells.     

Molecular cloning, genomic organization, and expression of a C-type (Manduca sexta-type) allatostatin preprohormone from Drosophila melanogaster

The insect allatostatins are a diverse group of neuropeptides that obtained their names by their inhibitory actions on the corpora allata (two endocrine glands near the insect brain), where they block the biosynthesis of juvenile hormone (a terpenoid important for development and reproduction). Chemically, the allatostatins can be subdivided into three different peptide groups: the large group of A-type (cockroach-type) allatostatins, which have the common C-terminal sequence Y/FXFGLamide; the B-type (cricket-type) allatostatins, which have the C-terminal sequence W(X(6))Wamide in common; and a single allatostatin that we now call C-type allatostatin that was first discovered in the moth Manduca sexta, and which has a nonamidated C terminus, and a structure unrelated to the A- and B-type allatostatins. We have previously cloned the preprohormones for the A- and B-type allatostatins from Drosophila melanogaster. Here we report on the cloning of a Drosophila C-type allatostatin preprohormone (DAP-C). DAP-C is 121 amino acid residues long and contains one copy of a peptide sequence that in its processed form has the sequence Y in position 4) from the Manduca sexta C-type allatostatin. The DAP-C gene has three introns and four exons and is located at position 32D2-3 on the left arm of the second chromosome. Northern blots show that the gene is strongly expressed in larvae and adult flies, but less in pupae and embryos. In situ hybridizations of larvae show that the gene is expressed in various neurons of the brain and abdominal ganglia and in endocrine cells of the midgut. This is the first publication on the structure of a C-type allatostatin from insects other than moths and the first report on the presence of all three types of allatostatins in a representative of the insect order Diptera (flies).     

Interaction between Manduca sexta allatotropin and manduca sexta allatostatin in the fall armyworm Spodoptera frugiperda

A peptide that strongly stimulates juvenile hormone (JH) biosynthesis in vitro by the corpora allata (CA) was purified from methanolic brain extracts of adult Spodoptera frugiperda. Using HPLC separation followed by Edman degradation and mass spectrometry, the peptide was identified as Manduca sexta allatotropin (Mas-AT). Treating the CA from adult S. frugiperda with synthetic Mas-AT (at 10(-6) M) caused an up to sevenfold increase in JH biosynthesis. The stimulation of JH synthesis was dose-dependent and reversible. Synthetic M. sexta allatostatin (Mas-AS) (10(-6) M) did not affect the spontaneous rate of JH secretion from CA of adult S. frugiperda, nor did any of the allatostatins of the Phe-Gly-Leu-amide peptide family tested. However, when CA had been activated by Mas-AT (10(-6) M), addition of synthetic Mas-AS (10(-6) M) reduced JH synthesis by about 70%. This allatostatic effect of Mas-AS on allatotropin-activated glands was also reversible. When CA were incubated in the presence of both Mas-AT (10(-6) M) and various concentrations of Mas-AS (from 10(-8) to 10(-5) M), the stimulation of JH-biosynthesis observed was inhibited in a dose-dependent manner. The experiments demonstrate a novel mechanism of allatostatin action. In S. frugiperda JH synthesis was inhibited only in those glands which had previously been activated by an allatotropin.     

Molecular cloning, genomic organization, and expression of a B-type (cricket-type) allatostatin preprohormone from Drosophila melanogaster

The insect allatostatins obtained their names because they block the biosynthesis of juvenile hormone (a terpenoid) in the corpora allata (two endocrine organs near the insect brain). Chemically, the allatostatins can be subdivided into three different peptide groups: the A-type allatostatins, first discovered in cockroaches, which have the C-terminal sequence Y/FXFGLamide in common; the B-type allatostatins, first discovered in crickets, which all have the C-terminal sequence W(X)(6)Wamide; and the C-type allatostatins, first discovered in the moth Manduca sexta, which have an unrelated and nonamidated C terminus. We have previously reported the structure of an A-type allatostatin preprohormone from the fruitfly Drosophila melanogaster. Here we describe the molecular cloning of a B-type prepro-allatostatin from Drosophila (DAP-B). DAP-B is 211 amino acid residues long and contains one copy each of the following putative allatostatins: AWQSLQSSWamide (drostatin-B1), AWKSMNVAWamide (drostatin-B2), 相似文献   

Allatostatins of the tiger prawn,Penaeus monodon (Crustacea: Penaeidea)

More than 40 peptides belonging to the -Y/FXFGL-NH(2) allatostatin superfamily have been isolated and identified from the central nervous system (CNS) of the tiger prawn, Penaeus monodon (Crustacea: Penaeidea). The peptides can be arranged in seven sub-groups according to the variable post-tyrosyl residue represented by Ala, Gly, Ser, Thr, Asn, Asp, and Glu. Two of the residues (Thr and Glu) have not been observed in this position previously in either insects or crustaceans. Also reported for the first time for allatostatins, two of the peptides are N-terminally blocked by a pyroglutamic acid residue. The yields of certain peptides with similar amino acid sequences to each other were, in some instances, very different. As an example, the yield of ANQYTFGL-NH(2) was 2pmol, compared with ASQYTFGL-NH(2), with a yield of 156 pmol. There are several possibilities to account for this. If, as in all species so far investigated, there is a single allatostatin gene in P. monodon, then it would appear that different sub-populations have contributed mutant forms of particular peptides to the extract. Another, less likely possibility is that this species has more than one allatostatin gene, producing a variable array of peptides albeit in different molar ratios. Several peptides were present apparently as a result of the loss of one or more residues at the N-terminus of a larger form, either due to N-terminal degradation or specific post-translational processing. The number of peptides identified exceeds that for any other insect or crustacean species previously investigated. None is identical to any of the 60-70 insect allatostatins so far identified, and only three are common to other crustaceans. Immunohistochemical study of the CNS of P. monodon, with the same antisera as used to monitor the purification, confirms the widespread nature and complexity of allatostatinergic neural pathways in arthropods. Thus, all neuromeres of the brain, and all except one of the ventral cord ganglia, possess allatostatin neurons and extensive areas of allatostatin-innervated neuropile. In addition to the cytological evidence that the allatostatins act as neurotransmitters, associated with tissues as varied as eyes and legs, their presence in neurohemal areas such as the sinus gland and the perineural sheath of the thoracic ganglia suggests a neuroendocrine function. As well as posing a challenge to physiologists assigning specific functions to the allatostatins, their extensive intra-species multiplicity, linked to their inter-species variability, also presents a complex problem to geneticists and evolutionists.