Cuticular Proteins in Insects and Crustaceans

Comparisons between crustacean and insect cuticles are hamperedby the paucity of cuticular protein sequences for the former.Sufficient complete sequences are available for insect cuticularproteins to allow recognition of conserved motifs and relationshipsamong proteins that reflect the type of cuticle from which theyhave been extracted. All five sequences from an arachnid andtwo of 14 from crustaceans have a motif found in the largestgroup of insect cuticular proteins. Numerous insights have beengained from studying insect cuticular proteins and their genes.These insights have been summarized in hopes of encouraginginterest in building on the foundations laid by Dorothy Skinnerwith the exoskeleton of Gecarcinus.     

Interactions Between Limb Regeneration and Molting in Decapod Crustaceans

Molting and regeneration of lost appendages are tightly-coupled,hormonally-regulated processes in decapod crustaceans. Precociousmolts are induced by eyestalk ablation, which reduces circulatingmolt-inhibiting hormone (MIH) and results in an immediate risein hemolymph ecdysteroids. Precocious molts are also inducedby autotomy of 5–8 walking legs; adult land crabs (Gecarcinuslateralis) molt 6–8 wk after multiple leg autotomy (MLA).Autotomy of one or more of the 1° limb buds (LBs) that formafter MLA before a critical period interrupts proecdysis until2° LBs re-regenerate and grow to the approximate size ofthose lost. Based on these observations, Skinner proposed thatlimb buds produce two factors that control proecdysial events.Limb Autotomy Factor–Anecdysis (LAF_an), produced by 1°LBs when at least five legs are autotomized, stimulates anecdysialanimals to enter proecdysis. Limb Autotomy Factor–Proecdysis(LAF_pro), produced by 2° LBs in premolt animals when atleast one 1° LB is autotomized, inhibits proecdysial processes.Initial characterizations suggest that LAF_pro is a MIH-likepolypeptide that inhibits the synthesis and secretion of ecdysteroidby the Y-organs.     

Physiological Control of Molting in Insects

SYNOPSIS. The initiation of a molting cycle in insects is neithera random nor a strictly periodic event. Insofar as molting canaccomplish different things under different circumstances, suchas a change in size or a change in form, it is reasonable toasume that the timing of a molt must be adapted to these functions.The onset of a metamorphic molt, in particular, must be preciselycontrolled because the onset of metamorphosis terminates thegrowth phase of a larva and establishes the body size of theadult insect. This aspect of the control of molting has receivedrelatively little attention and our knowledge of specific physiologicalmechanisms for the control of molt initiation is restrictedto three species: the blood-sucking bug, Rhodnius prolixus,the greater milkweed bug, Oncopeltus fasciatus, and the tobaccohornworm, Manduca sexta. The present review discusses the stateof our knowledge about the factors that render these insectscompetent to molt and about the stimuli that serve as a directtrigger for molting.     

水生甲壳类蜕皮发生过程及其影响因素的研究与进展

甲壳类的蜕皮发生贯穿甲壳类的整个生命周期,与其生长、发育及繁殖息息相关。相对于其他生物,甲壳类的生长是非连续性的,每次蜕皮过程都需要蜕去旧表皮,合成新表皮,同时体宽与体重在较短时间内实现一次跨越式增长。影响甲壳类的蜕皮发生的因素众多,本文重点从内源因子(蜕皮激素、蜕皮抑制激素、蜕皮激素受体与维甲类X受体、性腺发育)与外源因子(温度、光照、盐度、钙离子、饵料与污染物)两个方面概括总结了影响水生甲壳类蜕皮发生的关键因子及其国内外研究进展,分析了其关键因子对水生甲壳类蜕皮发生的影响机制,并尝试对水生甲壳类蜕皮发生当前存在的主要问题进行讨论。     

Enzymatic Hydrolysis of Proteins from Crustaceans of the Barents Sea

Enzymatic preparations from king crab hepatopancreas were shown to be capable, in principle, of producing protein hydrolysates. Hydrolysis of protein-containing waste of deep-water prawn and king crab occurs most successfully at pH 8.0–8.5 and 50–55°C for 5–6 h in the presence of 6 g enzyme per kg substrate. The total chemical composition of the hydrolysates, the molecular weight distributions of proteins and polypeptides, and the contents of free amino acids were studied in dry hydrolysates.     

病毒虫传相关蛋白

昆虫传播的植物病毒种类多、危害大。病毒虫传包括获毒、携毒与传毒3个阶段,是病毒、昆虫、植物互作的复杂过程,其中涉及到多种蛋白质。本文对与昆虫传播病毒有关的蛋白质进行了综述,并分析了这些蛋白质研究的科学意义与发展趋势,为植物病毒病的防治研究提供了一些思路。     

昆虫嗅觉相关蛋白及嗅觉识别机理研究概述

嗅觉是昆虫产生行为的基础之一,在长期进化的过程中昆虫形成了复杂的嗅觉系统,完成这一过程,需要有多种与嗅觉相关的蛋白参与,包括气味结合蛋白、化学感受蛋白、气味受体和感觉神经元膜蛋白等。了解昆虫感受外界信息的嗅觉机制可以帮助我们更好地理解昆虫识别配偶、天敌及寻找食物来源、产卵场地等行为特征,为进一步调控昆虫的行为、防控害虫侵袭、保护和利用有益昆虫奠定基础。本文综述了昆虫嗅觉相关的几类重要蛋白的生化特性和生理功能,并对昆虫气味分子的识别机制、气味分子在昆虫体内运输机制的最新研究进展进行了概述。     

Silkworm Midgut Proteins Interacting with a Hemolymph Protease Inhibitor,CI-8

CI-8 is the chymotrypsin inhibitor in hemolymph from the silkworm, Bombyx mori. It occurs in the midgut at the spinning stage of larva, but little information on the mechanism of its uptake in the midgut is available. We found that two polypeptides interacting with CI-8 are in the midgut membrane, and we purified them using a biotinylated CI-8, viz., p29 and p60, having molecular sizes of 29 kDa and 60 kDa respectively. The structures of p29 and p60 were examined by N-terminal amino acid sequencing and peptide mass mapping, including tryptic digestion. p29 was highly similar to the matured 19G1-30K lipoprotein from hemolymph, but p60 was novel. Purified p29 was recognized by anti-19G1-30K antibody, and was confirmed to be similar to 19G1-30K. The antibody also neutralized the CI-8 binding ability of p29 in the midgut membrane. p29 and p60 are perhaps proteinaceous factors involved in the uptake of CI-8 into the midgut through the membrane.     

Functional Analysis of Insect Molting Fluid Proteins on the Protection and Regulation of Ecdysis

Molting fluid accumulates between the old and new cuticles during periodical ecdysis in Ecdysozoa. Natural defects in insect ecdysis are frequently associated with melanization (an immunity response) occurring primarily in molting fluids, suggesting that molting fluid may impact immunity as well as affect ecdysis. To address this hypothesis, proteomic analysis of molting fluids from Bombyx mori during three different types of ecdysis was performed. Many proteins were newly identified, including immunity-related proteins, in each molting fluid. Molting fluids inhibited the growth of bacteria in vitro. The entomopathogenic fungi Beauveria bassiana, which can escape immune responses in feeding larvae, is quickly recognized by larvae during ecdysis, followed by melanization in molting fluid and old cuticle. Fungal conidia germination was delayed, and no hyphae were detected in the hemocoels of pharate instar insects. Molting fluids protect the delicate pharate instar insects with extremely thin cuticles against microorganisms. To explore the function of molting fluids in ecdysis regulation, based on protein similarity, 32 genes were selected for analysis in ecdysis regulation through RNAi in Tribolium castaneum, a model commonly used to study integument development because RNAi is difficult to achieve in B. mori. We identified 24 molting proteins that affected ecdysis after knockdown, with different physiological functions, including old cuticle protein recycling, molting fluid pressure balance, detoxification, and signal detection and transfer of molting fluids. We report that insects secrete molting fluid for protection and regulation of ecdysis, which indicates a way to develop new pesticides through interrupting insect ecdysis in the future.     

Immunity in Decapod Crustaceans

Immunity in the decapod crustaceans is surveyed. Types of immuneresponses include encapsulation, phagocytosis with or withoutthe aid of serum factors, bactericidins active with or withoutthe aid of hemocyte factors, hemagglutinins, hemolysins, agglutinins,and precipitins. Immunity to gaffkemia in Panulirus interruptusis also discussed.     

Hemolymph proteins in ticks

In comparison to insects and Crustacea, our knowledge of the predominant hemolymph proteins in ticks is minimal. The hemolymph protein most studied in ticks has been vitellogenin (Vg). Vg is synthesized by the tick fat body after female adults obtain a blood meal, is released into the hemolymph and is absorbed by developing oocytes as vitellin (Vn). Much of what we know about Vg is from studies of Vn. In general, the carbohydrate, lipid and amino acid composition is similar to insects except that in the tick, Vg contains heme, most likely from the digestion of host hemoglobin. In the American dog tick, Dermacentor variabilis, Vg is comprised of two native proteins and seven subunits on SDS-PAGE. Vg has been characterized in five tick species but the amino acid sequence is not yet available. Another predominant hemolymph protein, apparently a carrier protein (CP), has recently been studied in two tick species. This protein is found in the hemolymph of both male and females adults, in adult tissues outside of the hemolymph in some tick species, in coxal fluid of soft ticks and in whole body homogenates from eggs, larvae and nymphs. CP from the hard tick, D. variabilis, contains cholesterol, phospholipids, monoacylglycerides, triacylglycerides, free fatty acids, carbohydrate and heme. Under identical assay conditions, the analogous protein in the soft tick, Ornithodoros parkeri, did not contain heme. CP in the American dog tick consists of two subunits, one of which has 61% identity to the biliprotein, artemocyanin, from the fairy shrimp. CP is identical to a heme-lipoprotein (HeLp) from Boophilus microplus. The exact roles of CP and HeLp have not yet been fully determined, but they apparently are important in heme sequestration and as a storage depot for protein and lipid. Macroglobulin, lectin, antimicrobial, JH binding, JH esterase, and other tick hemolymph proteins are also discussed.     

MLT-10 Defines a Family of DUF644 and Proline-rich Repeat Proteins Involved in the Molting Cycle of Caenorhabditis elegans

The molting cycle of nematodes involves the periodic synthesis and removal of a collagen-rich exoskeleton, but the underlying molecular mechanisms are not well understood. Here, we describe the mlt-10 gene of Caenorhabditis elegans, which emerged from a genetic screen for molting-defective mutants sensitized by low cholesterol. MLT-10 defines a large family of nematode-specific proteins comprised of DUF644 and tandem P-X₂-L-(S/T)-P repeats. Conserved nuclear hormone receptors promote expression of the mlt-10 gene in the hypodermis whenever the exoskeleton is remade. Further, a MLT-10::mCherry fusion protein is released from the hypodermis to the surrounding matrices and fluids during molting. The fusion protein is also detected in strands near the surface of animals. Both loss-of-function and gain-of-function mutations of mlt-10 impede the removal of old cuticles. However, the substitution mutation mlt-10(mg364), which disrupts the proline-rich repeats, causes the most severe phenotype. Mutations of mlt-10 are also associated with abnormalities in the exoskeleton and improper development of the epidermis. Thus, mlt-10 encodes a secreted protein involved in three distinct but interconnected aspects of the molting cycle. We propose that the molting cycle of C. elegans involves the dynamic assembly and disassembly of MLT-10 and possibly the paralogs of MLT-10.     

Neurosecretion and Molting in Some Parasitic Nematodes

The adult female of Ascaris lumbricoides possesses a numberof nerve cells containing material which stains with paraldehyde-fuchsin.Among others, most of the primary sensory cells in the lipsare fuchsinophilic. Ascaris does not survive outside its host,so that it is impossible to ascribe a function to these cells. Phocanema decipiens possesses similar cells in the dorsal andventral ganglia which exhibit a cycle of secretion correlatedwith the burst of cytological activity which accompanies thedeposition of the new cuticle. Ligation experiments have demonstratedthat a new cuticle can be deposited in the absence of theseneurosecretory cells. Our most recent experiments suggest thatthe neurosecretory cells may control the release of leucineaminopeptidase in the excretory gland, a substance which isthought to be responsible for ecdysis.     

浓核病毒感染早期家蚕血液、中肠蛋白表达差异比较分析

为了探明在浓核病毒镇江株(BmDNV-ZJ)侵染早期,家蚕部分组织蛋白所产生的免疫抵抗性变化机制,本实验采用差异蛋白质组学技术研究分析了BmDNV-ZJ感染早期,家蚕的中肠、血液组织中特异性表达的差异蛋白.实验结果表明:在浓核病毒侵染初期,感受性家蚕的中肠组织受病毒感染而得到特异性表达的蛋白可能为丝氨酸蛋白酶抑制剂和巯基抗氧化酶蛋白,前者具有调控蛋白酶活性和细胞凋亡的功能,后者具有抗氧化的作用.血液组织受病毒感染诱导而产生的蛋白可能是丝裂原活化蛋白激酶和类抗氧化酶蛋白,前者具有调控细胞凋亡的功能,后者具有抗氧化、消除自由基作用.由于试验中所得的差异蛋白点很少,这表明在BmDNV-ZJ感染早期,蚕体对浓核病毒的感染而产生的反应很小.蚕体可通过被侵染的中肠组织(浓核病毒感染的靶部位)以及血液(免疫组织)共同产生一些抗氧化或调控细胞凋亡的酶蛋白等来抗击浓核病毒的侵染.     

Recognition of Non-self in Crustaceans

This paper reviews the current concepts of recognition of non-selfin crustaceans and relates these concepts to recognition inthe invertebrates in general. It focuses primarily on a decapodcrustacean, the blue crab(callinectes sapidus)and on resultsusing this animal as a model to study the clearance of virusesand xenogeneic proteins. Clearance studies indicate that bluecrabs possess a quasi-specific recognition system in the normalor "non-immunized"state. This system is capable of rapidly clearingforeign proteins and certain viruses from the circulation, andresults in concentration of such proteins into the gills andviruses into the hepatopancreas or gills. Although humoral factorswhich bind foreign proteins or neutralize viruses have beenisolated, transfusion exchange experiments involving depletionof circulating cells and/or humoral factors indicate that theblue crab does not require circulating hemocytes or humoralfactors for clearance of foreign proteins. These results suggestthat a population of fixed cells, possibly those in the gills,may be the critical component for recognition of foreign proteinsby normal crabs.     

Co-evolution Between Fishes and Crustaceans

In aquatic ecosystems fishes are, in general, the most important predators of crustaceans. This must not have been so at the beginning of the Mesozoic. Evolution of aquatic vertebrates led from heavy, benthic, microphagous animals to buoyant, agile, omnivorous ones. Key events of this evolution took place in fresh water, while most crustacean groups probably evolved in marine habitats. It is pointed out that until early Jurassic times the caridoid escape reaction obviously ensured the survival of eucarid species, while the later radiation suggests that selection favoured well-calcified and short-tailed species. It is suspected that the radiation of the marine Teleostei is a main cause of this evolution. This hypothesis is in accordance with the occurrence of the most archaic crustacean relict species in fish-free habitats.     

Hormonal Control of Molting and Reproduction in Ticks

SYNOPSIS. Among ticks there are two developmental and threereproductive patterns that correlate with taxonomic groupings(Argasidae, prostriate and metastriate Ixodidae). Feeding isa prerequisite for molting; feeding and mating are necessaryfor reproduction in all except a few parthenogenetic species.Growth and development in ticks and other chelicerates appearto be controlled by molting hormones (ecdysteroids), as theyare in insects and crustaceans. Ecdysone and 20-hydroxyecdysoneappear to be present in most or all of the major cheliceratetaxa. Epidermis is the site of ecdysone production and fat bodythe site of 20-hydroxylation in the argasid Ornithodoros parkeri,as is probably the case in all ticks. Ecdysteroids influenceearly stages of spermatogenesis by stimulation of DNA synthesisin spermatocytes, but controls for later stages of meiosis areunknown. A polypeptide (12,000 daltons) from male genital accessoryglands stimulates capacitation (maturation) of spermatids intosperm at the time of spermatid transfer to females. Knowledgeof control of egg development and oviposition is incomplete.Stimuli from the synganglion are necessary for completion ofoogenesis and two synganglial factors have been proposed. AnEgg Development Stimulation Factor (EDSF) in O. parkeri is synthesizedand/or released three to six days after feeding. VitellogenesisInducing Factor (VIF) in O. moubata is synthesized and/ or releasedwithin one hour after feeding. The VIF is hypothesized to impactan unidentified tissue which in turn produces a Fat Body StimulationFactor (FSF) that stimulates fat body to synthesize vitellogenin(Vg). Roles of ecdysteroids and juvenile hormones during eggdevelopment and oviposition are unclear.     

Aspects of Lipid Metabolism in Crustaceans

Lipid is the predominant organic reserve of many crustaceansand is important in the metabolism of many of these animals.Ingested lipid is digested by gastric lipase and apparentlyabsorbed into depot-lipid as rß-monoglycerides. Thevariation in the content and composition of the depot-lipidis a function of both the external environment and internalcontrol systems. Evidence suggests that lipids from marine organismscontain more long-chain polyunsaturated fatty acids than doesthe lipid of fresh water organisms which in turn have a highproportion of C₁₆ and C₁₈ fatty acids. The fatty-acid compositionof the sub-tropical land crab,Gecarcinns lateralis, resemblesthat of the fresh-water crustaceans. In addition, our studiesindicate that aspects of lipid metabolism may be under endocrinecontrol. The induction of premolt by destalking markedly increasesthe synthesis of lipid from metabolic precursors and its subsequentincorporation into the depot-lipid of the hepatopancreas. Inthe late premolt stages there is a decrease in the lipid contentof the hepatopancreas. This occurs as the lipid is mobilizedfrom the hepatopancreas to meet the energy demands of all thoseprocesses resulting in ecdysis. This sinusoidal variation inthe lipid metabolism of the hepatopancreas is influenced byan eyestalk factor (s).