昆虫气味认知的嗅觉神经结构及分子机制

大多数昆虫主要通过气味认知感知外界环境的变化,维持生命活动。探究昆虫气味认知的嗅觉系统神经结构及分子机制,对于完善气味认知神经生物学理论及利用其原理进行仿生学研究等有重要的科学意义。近年,关于昆虫气味认知科学研究有了很大的进展。本文从昆虫神经生物学的视角详细综述了近年关于昆虫气味认知的嗅觉神经结构、分子机制及气味信号的神经传导途径等方面的基本理论及最新研究成果。综述结果显示:昆虫对气味的认知是通过嗅觉神经系统的触角感器、触角叶(AL)、蕈形体(MB)等脑内多层信号处理神经结构来实现的。当外界气味分子进入触角感器内后,由感器内特定的气味识别蛋白(OBP)将气味分子运载到达嗅觉感受神经元(ORN)树突膜上的受体位点,气味分子与表达特定气味的受体(OR)结合产生电信号,并以动作电位的形式通过ORN的轴突传到脑内的触角叶。在触角叶经过嗅觉纤维球对气味信息选择性加工处理,再由投射神经元(PNs)将初步的识别和分类的气味信息传到蕈形体和外侧角(LH)等神经中枢,实现对气味的识别和认知。虽然,近年昆虫气味认知神经生物学的研究有了很大的进步,但是,我们认为目前的研究成果还不能完全阐明昆虫气味认知的神经机制,还有很多问题,例如,触角叶上众多的嗅觉纤维球是如何对嗅觉感受神经元传入的气味信息进行编码处理的?等有待进一步深入研究。为了搞清这些疑难问题,我们认为需要提高现有的实验技术水平,加强电生理学和分子神经生物学相结合的实验研究,从分子水平探究气味认知的神经机制可能是未来研究的热点。     

利用单感器记录与神经元示踪结合对棉铃虫主要性信息素感器内神经元投射的鉴定

【目的】鉴定雄性棉铃虫Helicoverpa armigera成虫触角性信息素感器嗅觉受体神经元的功能、形态及中枢投射路径。【方法】利用单感器记录技术记录棉铃虫嗅觉受体神经元对性信息素的反应,同时采用荧光染料作为示踪剂染色标记嗅觉受体神经元;使用免疫组织化学方法处理相应的脑组织,标记脑内触角叶的神经纤维球结构;用激光扫描共聚焦显微镜获取图像数据,使用图形软件ZEN和Amira 4.1.1进行三维结构重建。【结果】记录到雄性棉铃虫成虫触角上长毛形感器对主要性信息素成分Z11-16∶Ald产生明显的电生理反应,并成功染色标记了该感器内的嗅觉受体神经元。染色标记显示该感器内具有两个嗅觉受体神经元,其轴突通过触角神经分别投射触角叶内的云状体神经纤维球和普通神经纤维球。【结论】单感器记录与神经元示踪两技术结合能够用于鉴定昆虫触角嗅觉受体神经元的功能、形态和投射至神经纤维球的路径。与赖氨酸钴方法比较,使用荧光染料法进行神经元示踪,操作更简便,且易于进行三维空间分析,为调查棉铃虫其他嗅觉神经元的投射路径以明确外周气味受体感受与中枢系统的联系提供了有力技术支持。     

红缘天牛嗅觉感受器的类型及特点

利用扫描电镜对红缘天牛Asias halodendri（Pallas）的触角、下颚须和下唇须上嗅觉感受器的类型、超微形态结构及分布等特点进行观察。结果表明,红缘天牛触角、下颚须和下唇须上的嗅觉感器共有6类14种。触角上感器共有2类9种,即毛形感器1种,锥形感器8种;触角上毛形感器和Ⅱ型锥形感器长度在雌雄间有显著性差异;Ⅲ型、Ⅶ型和Ⅷ型锥形感器为雌性触角上所特有;Ⅵ型锥形感器仅见于雄性触角;触角上的感受器总数雌性数量多于雄性,背面略多于腹面和侧面。下颚须及下唇须上嗅感器共4类5种,即栓锥形感器2种、板形感器、钟形感器和边缘凹槽形感器各1种;雌性的边缘凹槽形感器明显高于雄性;下颚须和下唇须上雄性感器总数较多,腹面数量多于背面。     

蚊虫搜寻吸血寄主和产卵行为的调节因子及相关嗅觉机理

嗅觉在蚊虫的吸血寄主搜寻、产卵和糖源搜寻行为中起决定作用,而在交配行为中的作用并不清楚。本文系统全面地综述了近20年来蚊虫化学生态学和嗅觉识别的分子机理的研究。蚊虫的触角、下颚须和口喙上的嗅觉感器感觉环境中释放的各种挥发性化合物。气味分子与嗅觉气味结合蛋白和气味受体的结合所启动的一系列生化反应产生神经动作电位。蚊虫嗅觉神经元编码气味中化合物的组成、浓度及其暂时瞬间的浓度变化和空间分布。吸血前后神经元的活性在数量和质量上有变化,反映了蚊虫在搜寻吸血寄主和产卵行为上的调节。在吸血寄主搜寻中,人体和动物释放的二氧化碳、乳酸以及其他气味协同引诱蚊虫向目标气味源定向飞行,最后找到吸血寄主。而成熟产卵雌蚊是利用产卵场所释放的腐烂气味寻找适宜的产卵场所,一些蚊虫卵、幼虫或蛹分泌的产卵信息素引诱和刺激雌蚊产卵,并与产卵生境气味起协同作用。植物气味尤其是花香味引诱蚊虫找到蜜源。驱避剂也是直接或间接通过嗅觉起作用,一些驱蚊剂由于阻断嗅觉反应而抑制蚊虫的定向飞行。从植物、动物或人体以及产卵场所释放的气味中有望找到有效的引诱和驱避化合物。对蚊虫嗅觉识别机理的认识将使我们开发出有效的蚊虫诱捕技术,进而应用于种群监测和控制。     

小地老虎雌蛾触角及幼虫头部感受器扫描电镜观察

【目的】为了解小地老虎Agrotis ypsilon(Rottemberg)雌蛾触角及幼虫头部感受器的种类、形态、数量和分布。【方法】利用扫描电镜技术观察了小地老虎雌蛾触角及5龄幼虫头部形态及触角和口器上的感受器。【结果】小地老虎雌蛾触角上共分布有5种类型的感受器,分别为毛形感器、刺形感器、腔锥形感器、耳形感器、锥形感器。5龄幼虫头部呈椭圆形,口器下口式,为咀嚼式口器,头部可见触角、单眼、蜕裂线、刚毛、唇基、额、上唇、上颚、下颚、下颚须、下唇、下唇须及吐丝器等结构;触角上着生1个毛形感器和4个锥形感器;上颚分布有1个毛形感器和1个刺形感器;在下颚及下颚须上共分布有3个毛形感器、3个刺形感器、4个锥形感器和1个栓锥形感器;下唇须上有1个栓锥形感器和1个锥形感器;吐丝器前方两侧有1对刺形感器。【结论】小地老虎雌蛾触角上的感受器比幼虫头部的多,推测雌蛾利用它们来完成寄主植物和产卵环境选择;幼虫口器附肢上的感受器具有味觉和嗅觉功能,幼虫利用它们判断食物的种类和适合性。     

锈翅蚁蛉头部附器感器的扫描电镜观察

利用扫描电镜对锈翅蚁蛉Myrmeleon ferrugineipennis Bao&Wang雌雄成虫头部触角及口器感器的形态进行观察,描述了感器的种类、数量和分布,以期解析其取食机制。结果表明:锈翅蚁蛉触角上存在10种感器,即毛形感器、锥形感器、刺形感器、腔形感器、钟状感器、鳃形感器、耳形感器、盘形感器、舌形感器和Bhm氏鬃毛,其中毛形感器有3种亚型,数量最多;耳形感器、腔形感器和钟状感器仅在雌成虫触角上发现,而舌形感器和鳃形感器仅在雄成虫触角上发现;在锈翅蚁蛉触角鞭节近末端扁平匙状处各有1枚盘形感器,其形状和位置在雌雄虫上有差异。鳃形感器和盘形感器在已有的昆虫感器研究中未见报道,是新发现的昆虫触角感器。下颚须、下唇须上均发现锥形感器,下唇须上的数量多于下颚须;此外,下颚须上还存在钟状感器。     

本期重点推介

正研究棉铃虫Helicoverpa armigera嗅觉感受的神经机制,可为进一步研发棉铃虫引诱剂技术提供理论依据。单感器记录与神经元示踪技术相结合是研究昆虫嗅觉受体神经元的功能及其投射的神经纤维球的重要方法。为了鉴定棉铃虫雄性成虫触角性信息素感器嗅觉受体神经元的功能、形态及中枢投射路径,河南农业大学植物保护学院马百伟和赵新成及中国农业科学院植物保护研究所王桂荣等以雄性棉铃虫成虫为试虫,利用单感器记录技术记录其嗅觉受体神经元     

昆虫触角叶的结构

触角叶是昆虫脑内初级嗅觉中心,通过触角神经与触角联系。触角叶主要由嗅觉受体神经元、局域中间神经元、投射神经元和远心神经元构成。这些神经元的形态多样,其形态变化与其功能和昆虫嗅觉行为相关。这些神经元在触角叶内交织形成神经纤维网,在突触联系紧密的地方形成纤维球,纤维球通常排列在触角叶外周。通常,昆虫触角叶内纤维球的数量、大小和位置相对固定,并且几乎每个小球都可以被识别和命名。不同种类、性别和品级的昆虫中,纤维球的数量、大小和排列方式各不相同。触角叶结构神经元组成和纤维球的多样性,与各种昆虫嗅觉行为的特异性相对应。     

灰茶尺蠖成虫触角及幼虫头部感器超微结构

【目的】明确灰茶尺蠖Ectropis grisescens成虫触角及幼虫头部感器的种类、形态、数量和分布,以探讨灰茶尺蠖的行为机制。【方法】利用扫描电镜技术观察灰茶尺蠖雌、雄成虫触角和5龄幼虫头部感器的超微结构。【结果】灰茶尺蠖成虫触角上分布有8种感器,分别是栓锥形感器、耳形感器、毛形感器(Ⅰ-Ⅳ)、B?hm氏鬃毛、腔锥形感器(Ⅰ和Ⅱ)、鳞形感器、锥形感器(Ⅰ和Ⅱ)和刺形感器。其中,栓锥形感器仅分布在雌蛾触角上,耳形感器、毛形感器(STⅠ-Ⅲ)仅分布在雄虫触角上。5龄幼虫触角上着生1个栓锥形感器、1个锥形感器和2个刺形感器;上唇着生有6对刺形感器,内唇着生有3对刺形感器和1对指形感器;上颚基部外侧着生有2个刺形感器;下颚及下颚须着生有5个刺形感器、9个锥形感器和2个栓锥形感器;下唇须着生有1个锥形感器和1个刺形感器;吐丝器前端着生有1对刺形感器。【结论】灰茶尺蠖雌、雄成虫触角感器存在性二型性,且雄虫上感器种类和数量较多,据此推测雄虫感受寄主植物或性信息素的能力较强;幼虫头部感器具有嗅觉和味觉功能,在其判断食物的种类和适应性等生态行为中发挥重要作用。     

果蝇幼虫气味编码的分子基础

全球农业经济的损失主要来自昆虫幼虫的取食,而昆虫幼虫的取食主要是依靠气味介导的嗅觉作用。耶鲁大学的昆虫嗅觉神经生物学家对黑腹果蝇Drosophila melanogaster幼虫嗅觉分子基础进行了研究。RT—PCR扩增出23个气味受体基因,其中13个基因,同成虫触角和下颚须内的气味受体基因相同,     

Central projections of olfactory receptor neurons from single antennal and palpal sensilla in mosquitoes

In insects, olfactory receptor neurons (ORNs) are located in cuticular sensilla, that are present on the antennae and on the maxillary palps. Their axons project into spherical neuropil, the glomeruli, which are characteristic structures in the primary olfactory center throughout the animal kingdom. ORNs in insects often respond specifically to single odor compounds. The projection patterns of these neurons within the primary olfactory center, the antennal lobe, are, however, largely unknown.We developed a method to stain central projections of intact receptor neurons known to respond to host odor compounds in the malaria mosquito, Anopheles gambiae. Terminal arborizations from ORNs from antennal sensilla had only a few branches apparently restricted to a single glomerulus. Axonal arborizations of the different neurons originating from the same sensillum did not overlap.ORNs originating from maxillary palp sensilla all projected into a dorso-medial area in both the ipsi- and contralateral antennal lobe, which received in no case axon terminals from antennal receptor neurons. Staining of maxillary palp receptor neurons in a second mosquito species (Aedes aegypti) revealed unilateral arborizations in an area at a similar position as in An. gambiae.     

A large-scale model of the locust antennal lobe

The antennal lobe (AL) is the primary structure within the locust’s brain that receives information from olfactory receptor neurons (ORNs) within the antennae. Different odors activate distinct subsets of ORNs, implying that neuronal signals at the level of the antennae encode odors combinatorially. Within the AL, however, different odors produce signals with long-lasting dynamic transients carried by overlapping neural ensembles, suggesting a more complex coding scheme. In this work we use a large-scale point neuron model of the locust AL to investigate this shift in stimulus encoding and potential consequences for odor discrimination. Consistent with experiment, our model produces stimulus-sensitive, dynamically evolving populations of active AL neurons. Our model relies critically on the persistence time-scale associated with ORN input to the AL, sparse connectivity among projection neurons, and a synaptic slow inhibitory mechanism. Collectively, these architectural features can generate network odor representations of considerably higher dimension than would be generated by a direct feed-forward representation of stimulus space.     

Physiology and antennal lobe projections of olfactory receptor neurons from sexually isomorphic sensilla on male <Emphasis Type="Italic">Heliothis virescens</Emphasis>

The neurophysiology and antennal lobe projections of olfactory receptor neurons (ORNs) within sexually isomorphic short trichoid sensilla of male Heliothis virescens (Noctuidae: Lepidoptera) were investigated using cut-sensillum recording and cobalt-lysine staining. A total of 202 sensilla were sorted into 14 possible sensillar categories based on odor responses and physiology of ORNs within. Seventy-two percent of the sensilla identified contained ORNs stimulated by conspecific odors. In addition, a large number of ORNs were specifically sensitive to ß-caryophyllene, a plant-derived volatile (N = 41). Axons originating from ORNs associated with individual sensilla were stained with cobalt lysine (N = 67) and traced to individual glomeruli in the antennal lobe. ORNs with responses to female sex pheromone components exhibited similar axonal projections as those previously described from ORNs in long sensilla trichodea in male H. virescens. Antennal lobe axonal arborizations of ORNs sensitive to hairpencil components were also located in glomeruli near the base of the antennal nerve, whilst those sensitive to plant odorants projected to more medial glomeruli. Comparisons with ORNs described from female H. virescens supports the notion that glomeruli at the base of the antennal nerve are associated with conspecific and interspecific odorants, whereas those located medially are associated with plant volatiles.     

Molecular, anatomical, and functional organization of the Drosophila olfactory system

BACKGROUND: Olfactory receptor neurons (ORNs) convey chemical information into the brain, producing internal representations of odors detected in the periphery. A comprehensive understanding of the molecular and neural mechanisms of odor detection and processing requires complete maps of odorant receptor (Or) expression and ORN connectivity, preferably at single-cell resolution. RESULTS: We have constructed near-complete maps of Or expression and ORN targeting in the Drosophila olfactory system. These maps confirm the general validity of the "one neuron--one receptor" and "one glomerulus--one receptor" principles and reveal several additional features of olfactory organization. ORNs in distinct sensilla types project to distinct regions of the antennal lobe, but neighbor relations are not preserved. ORNs grouped in the same sensilla do not express similar receptors, but similar receptors tend to map to closely appositioned glomeruli in the antennal lobe. This organization may serve to ensure that odor representations are dispersed in the periphery but clustered centrally. Integrated with electrophysiological data, these maps also predict glomerular representations of specific odorants. Representations of aliphatic and aromatic compounds are spatially segregated, with those of aliphatic compounds arranged topographically according to carbon chain length. CONCLUSIONS: These Or expression and ORN connectivity maps provide further insight into the molecular, anatomical, and functional organization of the Drosophila olfactory system. Our maps also provide an essential resource for investigating how internal odor representations are generated and how they are further processed and transmitted to higher brain centers.     

Olfactory Coding in Antennal Neurons of the Malaria Mosquito, Anopheles gambiae

Olfactory receptor neurons (ORNs) in the antenna of insects serve to encode odors in action potential activity conducted to the olfactory lobe of the deuterocerebrum. We performed an analysis of the electrophysiological responses of olfactory neurons in the antennae of the female malaria mosquito Anopheles gambiae s.s. and investigated the effect of blood feeding on responsiveness. Forty-four chemicals that are known to be present in human volatile emanations were used as odor stimuli. We identified 6 functional types of trichoid sensilla and 5 functional types of grooved-peg sensilla (GP) based on a hierarchical cluster analysis. Generalist ORNs, tuned to a broad range of odors, moderate specialist ORNs and 2 ORNs tuned to only one odor were identified in different sensilla types. Neurons in GP were tuned to more polar compounds including the important behavioral attractant ammonia and its synergist L-lactic acid, responses to which were found only in GP. Combinatorial coding is the most plausible principle operating in the olfactory system of this mosquito species. We document for the first time both up- and downregulation of ORN responsiveness after blood feeding. Modulation of host-seeking and oviposition behavior is associated with both qualitative and quantitative changes in the peripheral sensory system.     

东北大黑鳃金龟嗅感器超微结构

利用扫描电镜和透射电镜对东北大黑鳃金龟Holotrichia diomphalia成虫触角嗅感器进行超微结构研究。结果表明： 其嗅感器集中于触角鳃片上,着生在表皮内陷形成的凹腔里。嗅感器包括锥形感器和板形感器两种,锥形感器根据锥体形状的差异可分为4种类型,板形感器根据盘体形状的不同可分为5种类型。嗅感器表皮为单壁,壁上具有微孔和孔道微管。嗅感器内神经元的数目并不一致,1～3个不等。雄性触角鳃片的长度长于雌性触角鳃片,并且雄性触角嗅感器的总数远远多于雌性,其中雄性板形感器的数目与雌性差异不大,但雄性锥形感器的数目却远远的多于雌性,几乎是雌性的9倍。由此推测锥形感器是感受性信息素的感器,而板形感器用于感受植物气味。     

Olfactory receptors on the antennae of the malaria mosquito Anopheles gambiae are sensitive to ammonia and other sweat-borne components

Electrophysiological studies on female An. gambiae s.s. mosquitoes revealed a receptor neuron within a subpopulation of the antennal grooved-peg sensilla sensitive to the odour of incubated sweat, but not responding to fresh sweat. This receptor neuron was sensitive to ammonia as well, a sweat-borne component which attracts female An. gambiae in a windtunnel bioassay. Neurons innervating a different subpopulation of grooved-peg sensilla did not show a response to incubated sweat. In the latter sensilla, however, one type of neuron responded to water or water containing solutions, while another receptor neuron was inhibited when stimulated with dry air, ether or ethanol. Neurons innervating sensilla trichodea, a more abundant antennal type of olfactory sensillum, did not respond to fresh or incubated sweat at the doses offered. However, receptor neurons within the sensilla trichodea responded with excitation to several sweat-borne components. A subpopulation of the sensilla trichodea was innervated by neurons sensitive to geranyl acetone. A second subpopulation housed receptor neurons sensitive to indole. 3-Methyl-1-butanol and 6-methyl-5-hepten-2-one evoked excitation of receptor neurons within both subpopulations of sensilla trichodea. Neurons were most sensitive to indole and geranyl acetone with a threshold of 0.01%. These findings are discussed in the context of host-seeking behaviour.     

Unusual pheromone receptor neuron responses in heliothine moth antennae derived from inter-species imaginal disc transplantation

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons housed in sensilla trichodea along the adult antennae arising from transplantation of the antennal imaginal discs between larval male Helicoverpa zea and Heliothis virescens. The olfactory receptor neurons from the majority of type C sensilla sampled on transplanted antennae displayed response characteristics consistent with those of the species that donated the antennae. However, some of the sensilla type C sampled in either transplant type contained olfactory receptor neurons that responded in a manner typical of the recipient species or other neurons that have not previously been found in the type C sensilla of either species. The single-cell data help to explain behavioral results showing that some transplant males do fly upwind to both species' pheromone blends, an outcome not expected based on known antennal sensory phenotypes. Our results suggest that host tissue can influence antennal olfactory receptor neuron development, and further that because of a common phylogenetic ancestry the donor tissue has the genetic capability to produce a variety of sensillar and receptor types.     

Morphological and ultrastructural characterization of olfactory sensilla in Drosophila suzukii: Scanning and transmission electron microscopy

Drosophila suzukii is a serious horticultural and quarantine pest, damaging various berry crops. Although the active use of olfactory communication in D. suzukii is well-known, their olfactory sensory system has not been comprehensively reported. Therefore, the present study was carried out to understand the morphology, distribution and ultrastructure of olfactory sensilla present in the antennae and maxillary palps of D. suzukii, through scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The olfactory sensilla on the antennae of D. suzukii in both sexes could be classified into three major morphological types, basiconic, trichoid and coeloconic sensilla, according to their shapes. The antennal basiconic sensilla were further divided into three subtypes and the antennal trichoid sensilla into two subtypes, respectively, according to the size of individual sensillum. In contrast to the antennal olfactory sensilla showing diverse morphology, basiconic sensilla was the only type of olfactory sensilla in the maxillary palps of D. suzukii. The basiconic sensilla in the maxillary palps could be further classified into three subtypes, based on their size. Our SEM and TEM observations indicated that multiple nanoscale pores are present on the surface of all types of olfactory sensilla in the antennae and maxillary palps, except coeloconic sensilla. The difference in the morphological types and the distribution of olfactory sensilla suggests that their olfactory functions are different between antennae and maxillary palps in D. suzukii. The results of this study provide useful information for further studies to determine the function of olfactory sensilla in D. suzukii and to understand their chemical communication system.