脊索动物嗅觉受体基因命名法的发展

嗅觉受体属于G蛋白偶联受体家族,在脊索动物的整个生命周期中都扮演着至关重要的角色。与其他多数基因家族不同,嗅觉受体家族是一个成员数量庞大的超基因家族,为它们合乎逻辑的命名可以更好地对该家族进行描述、分析和讨论,也可以为机器学习程序从庞大的嗅觉受体数据库自动构建相应的蛋白结构和功能知识库提供语义信息。由于脊索动物嗅觉受体演化速度很快、基因数量庞大、假基因比率高、在物种及染色体上分布差异巨大等多方面的原因,给嗅觉受体基因合理的命名较为困难。三十多年来,伴随着嗅觉受体研究领域的发展,嗅觉受体基因命名法也经历了多次迭代,在每个阶段都发挥着积极的作用。随着测序技术和生物信息学算法工具的发展,随之而来的是新注释的海量的嗅觉受体基因,这使已有的嗅觉受体基因命名法变得越来越难以适应大数据挖掘和知识工程的系统开发,因此迫切需要一个能满足当下需求的嗅觉受体基因命名法。     

基因组水平蝙蝠嗅觉受体基因的分子进化

翼手目动物（俗称蝙蝠）的食性分化显著,不同食性的蝙蝠具有显著不同的嗅球大小。为了研究嗅觉是否影响了蝙蝠食性的进化,我们利用网上已公布的10种蝙蝠基因组的数据,通过同源比对的方法鉴定出所有的嗅觉受体基因,并进行嗅觉受体基因亚家族的分类,进而比较嗅觉受体基因亚家族的数目差异。结果显示,蝙蝠的嗅觉受体基因与其它哺乳动物一样,都可以分为13个单系起源的亚家族;在Yinpterochiroptera亚目中,OR1/3/7、OR2/13、OR5/8/9等3个嗅觉受体亚家族在食果蝙蝠中均发生了不同程度的扩张,基因数目显著地多于食虫蝙蝠,提示嗅觉在食果蝙蝠取食过程中具有重要的作用。因此,本研究在基因组水平上重现了蝙蝠嗅觉受体基因的进化历史,揭示了3个嗅觉受体基因亚家族的功能可能与食果蝙蝠的食性相关,突出了嗅觉对动物食性的重要作用.     

腺苷酸环化酶3缺失下调小鼠嗅觉受体基因表达北大核心CSCD

主要嗅觉表皮组织(MOE)是哺乳动物感知气味分子的重要器官,气味诱导是嗅觉受体神经元(ORN)活动的起点,嗅觉受体(OR)结合气味分子后通过环腺苷酸(c AMP)信号通路向下游传递信号。腺苷酸环化酶3(AC3)是此通路中的重要分子。为了探讨AC3缺失对小鼠MOE内ORs基因表达的影响,本文以AC3敲除型小鼠(AC3-/-)和野生型小鼠(AC3+/+)为材料,采用荧光定量PCR(qRT-PCR)、荧光原位杂交(FISH)技术分析了部分ORs基因及与其相关因子在MOE中的表达。qRT-PCR表明,3月龄AC3-/-小鼠MOE中嗅觉受体Olfr15、Olfr16、Olfr533、Olfr536、Olfr1507和Olfr642的表达量均显著下降。出生后PND7、PND30和PND90三个不同发育时期的AC3-/-小鼠MOE原位杂交显示,嗅觉受体Olfr15、Olfr536和Olfr1507表达的细胞数目均减少。进一步qRTPCR分析发现,3月龄AC3-/-小鼠嗅觉受体相关因子Rtp1、Rtp2、Reep1、Lhx2、Emx2和Ric-8b的表达也均发生显著下调。由此推测,AC3缺失导致的ORs及其相关因子的表达下调可能是嗅觉行为障碍的原因之一。     

腺苷酸环化酶3缺失下调小鼠嗅觉受体基因表达

主要嗅觉表皮组织（MOE）是哺乳动物感知气味分子的重要器官,气味诱导是嗅觉受体神经元（ORN）活动的起点,嗅觉受体（OR）结合气味分子后通过环腺苷酸（cAMP）信号通路向下游传递信号。腺苷酸环化酶3（AC3）是此通路中的重要分子。为了探讨AC3缺失对小鼠MOE内ORs基因表达的影响,本文以AC3敲除型小鼠(AC3-/-)和野生型小鼠(AC3+/+)为材料,采用荧光定量PCR（qRT-PCR）、荧光原位杂交（FISH）技术分析了部分ORs基因及与其相关因子在MOE中的表达。qRT-PCR表明,3月龄AC3-/-小鼠MOE中嗅觉受体 Olfr15、Olfr16、Olfr533、Olfr536、Olfr1507和Olfr642的表达量均显著下降。出生后PND7、PND30和PND90 三个不同发育时期的AC3-/-小鼠MOE原位杂交显示,嗅觉受体Olfr15、Olfr536和Olfr1507表达的细胞数目均减少。进一步qRT-PCR分析发现,3月龄AC3-/-小鼠嗅觉受体相关因子Rtp1、Rtp2、Reep1、Lhx2、Emx2和Ric-8b的表达也均发生显著下调。由此推测,AC3缺失导致的ORs及其相关因子的表达下调可能是嗅觉行为障碍的原因之一。     

2004年度诺贝尔生理学和医学奖简介

2004年诺贝尔生理学和医学奖颁发给两位美国科学家理查德·阿克塞尔(Richard Axel)和琳达·巴克(Linda Buck).他们发现嗅觉系统中一个大家族基因,这一大家族基因可以表达等量的嗅觉受体类型.这些受体位于鼻腔上皮的嗅觉神经元上,以检测不同的气味分子.     

昆虫非典型嗅觉受体Orco的功能和分子结构研究进展

嗅觉受体是参与昆虫嗅觉识别过程的一类重要蛋白。在昆虫的众多嗅觉受体中, 有一类受体明显不同于其他受体, 被称为Orco。该受体基因在不同昆虫种间高度保守, 且表达广泛。Orco在昆虫嗅觉识别过程中发挥关键作用。采用基因突变或RNAi等技术使Orco基因沉默后, 昆虫会出现严重的嗅觉缺陷, 但Orco本身不与气味配体结合, 它与传统嗅觉受体形成复合体Or-Orco, 促进传统嗅觉受体在神经元树突膜上的定位并维持其稳定性, 提高传统嗅觉受体对气味反应的效率。昆虫嗅觉受体的结构与脊椎动物的G蛋白偶联受体相似, 均有7个跨膜区, 但二者的膜拓扑结构相反, 昆虫嗅觉受体的N末端位于细胞质膜内, C末端在细胞质膜外, Orco与传统嗅觉受体通过保守的C末端区域相互作用形成一种新型的配体门控离子通道--Or-Orco复合体。阐明Orco在昆虫嗅觉识别中的功能机制, 可为开创基于昆虫嗅觉行为干扰的新的害虫防治措施提供基础。     

昆虫嗅觉受体的研究进展

昆虫的嗅觉对昆虫的栖息地选择、觅食、群集、趋避、繁殖以及信息传递等行为具有重要的影响。对昆虫嗅觉机理的深入研究和嗅觉信号传导途径的完整阐述,是探索农业害虫的专一性防治的基础。嗅觉受体(olfactory receptors,Ors)是G蛋白偶联受体(G protein-coupled receptor)的一种,是嗅觉系统的关键成分。近年来嗅觉受体的研究日益受到关注。本文对昆虫嗅觉的基本过程、基因结构和表达调控特征、蛋白分子结构、生理功能、分布部位和相关配体的研究等进行了综述。     

拉布拉多犬嗅觉受体CfOLF4基因的克隆及序列分析

以常规PCR技术扩增警用拉布拉多犬(Canis familiaris)的嗅觉受体CfOLF4基因长879bp的目的片段,构建pMD-CfOLF4载体进行T-A克隆。重组质粒测序后与GenBank公布的犬CfOLF4基因进行序列分析比较,旨在了解拉布拉多犬嗅觉受体CfOLF4基因遗传的同源性和变异特性。     

鱼类嗅觉系统和性信息素受体的研究进展

鱼类嗅觉系统包括外部嗅觉器官、嗅神经和嗅球三个部分.嗅觉器官也称为嗅囊,由嗅上皮和髓质组成.气味物质的化学信息主要由嗅上皮上随机分布的嗅觉感受神经元感知,通过嗅神经将嗅觉信息传递到嗅球,嗅球在空间上有不同的功能分区,嗅觉信息经过嗅球各分区整合后分别传入端脑,发挥其生理功能.性信息素在鱼类生殖过程中的作用是通过嗅觉系统来完成的,其中嗅觉感受神经元上的性信息素受体起着重要作用.鱼类性信息素受体的研究主要从两个方面入手,一是从低浓度特异的性信息素引起嗅觉器官电生理反应或行为反应入手,寻找特异的性信息素受体;二是参照哺乳动物嗅觉受体的研究结果,从嗅觉受体基因遗传保守性入手,研究鱼类性信息素受体的结构与功能.     

锤胁跷蝽触角转录组分析

为了获得锤胁跷蝽(Yemma signatus Hsiao)触角基因信息,本研究使用Illumina HiSeq TM 4 000高通量测序技术对锤胁跷蝽成虫触角进行转录组测序和生物学信息分析。共获得61 080 938条clean reads,包括9.16 G (GenBank登录号:SRR3348966)。拼接115 491条unigenes,平均长度为578 bp,N50为863 bp。在7大数据库中注释到46 224条unigenes,其中在NR数据库中注释的基因最多,为32 842 (28.43%)条,与内华达古白蚁(Zootermopsis nevadensis)相似度最高(9.0%)。转录组数据分析鉴定出125个与嗅觉相关的基因,其中66个嗅觉受体、11个味觉受体、1个离子型受体、3个感觉神经元膜蛋白、30个气味结合蛋白和14个化学感受蛋白基因。本研究首次获得锤胁跷蝽触角转录组数据,并鉴定出嗅觉相关基因,本研究为今后利用嗅觉基因靶标防治害虫提供了分子生物学信息数据。     

Molecular biological research on olfactory chemoreception in fishes

This review describes recent molecular biological research on olfactory chemoreception in fishes. The recent rapid development of molecular biological techniques has provided new valuable information on the main and vomeronasal olfactory receptor (OR) genes, the axonal projection from ciliated, microvillous and crypt-olfactory receptor cells to the olfactory bulb, properties of odorant substances and olfactory imprinting and homing in salmon. Many important questions, however, remain unanswered on functional differences among OR genes, on ligand binding to each OR and on the molecular biological mechanisms underlying olfactory imprinting and homing in salmon. Olfactory chemoreception is believed to be the oldest sensory cue for both animal survival and adaptation to various different environments. Further intensive molecular biological research on olfactory memory formation and remembrance should be carried out to clarify the fundamental process of olfactory chemoreception in fishes.     

Avian olfactory receptor gene repertoires: evidence for a well-developed sense of smell in birds?

Among vertebrates, the sense of smell is mediated by olfactory receptors (ORs) expressed in sensory neurons within the olfactory epithelium. Comparative genomic studies suggest that the olfactory acuity of mammalian species correlates positively with both the total number and the proportion of functional OR genes encoded in their genomes. In contrast to mammals, avian olfaction is poorly understood, with birds widely regarded as relying primarily on visual and auditory inputs. Here, we show that in nine bird species from seven orders (blue tit, Cyanistes caeruleus; black coucal, Centropus grillii; brown kiwi, Apteryx australis; canary, Serinus canaria; galah, Eolophus roseicapillus; red jungle fowl, Gallus gallus; kakapo, Strigops habroptilus; mallard, Anas platyrhynchos; snow petrel, Pagodroma nivea), the majority of amplified OR sequences are predicted to be from potentially functional genes. This finding is somewhat surprising as one previous report suggested that the majority of OR genes in an avian (red jungle fowl) genomic sequence are non-functional pseudogenes. We also show that it is not the estimated proportion of potentially functional OR genes, but rather the estimated total number of OR genes that correlates positively with relative olfactory bulb size, an anatomical correlate of olfactory capability. We further demonstrate that all the nine bird genomes examined encode OR genes belonging to a large gene clade, termed gamma-c, the expansion of which appears to be a shared characteristic of class Aves. In summary, our findings suggest that olfaction in birds may be a more important sense than generally believed.     

Signatures of selection in the human olfactory receptor OR5I1 gene

The human olfactory receptor (OR) repertoire is reduced in comparison to other mammals and to other nonhuman primates. Nonetheless, this olfactory decline opens an opportunity for evolutionary innovation and improvement. In the present study, we focus on an OR gene, OR5I1, which had previously been shown to present an excess of amino acid replacement substitutions between humans and chimpanzees. We analyze the genetic variation in OR5I1 in a large worldwide human panel and find an excess of derived alleles segregating at relatively high frequencies in all populations. Additional evidence for selection includes departures from neutrality in allele frequency spectra tests but no unusually extended haplotype structure. Moreover, molecular structural inference suggests that one of the nonsynonymous polymorphisms defining the presumably adaptive protein form of OR5I1 may alter the functional binding properties of the OR. These results are compatible with positive selection having modeled the pattern of variation found in the OR5I1 gene and with a relatively ancient, mild selective sweep predating the "Out of Africa" expansion of modern humans.     

长江刀鲚mor-4k13基因的分离鉴定及表达分析

分布于长江的刀鲚(Coilia nasus)具有洄游和定居两个生态型,生殖洄游是区别两者的主要表征。为了探索嗅觉受体(OR)基因是否参与了刀鲚的生殖洄游过程,本文采用RACE技术从洄游型刀鲚中获得了mor-4k13基因,该基因全长1 098 bp,编码区长963 bp,单外显子结构,可编码320个氨基酸。预测表明,mor-4k13基因编码的蛋白质,为7个疏水性的α-螺旋跨膜结构,属G-蛋白偶联受体,有胆固醇和油酸两个配体。MOR-4K13蛋白与已报道的其他鱼类OR蛋白的同源性在40%~68%之间,其中,与近缘种大西洋鲱(Clupea harengus)嗅觉受体蛋白同源性高达68%。采用Real-time PCR方法对10个组织或器官所作的荧光定量分析显示,mor-4k13基因在定居型刀鲚嗅囊和性腺中高表达,在肌肉、眼球、胃壁、肝和鳃中低表达,心肌中几乎不表达。mor-4k13基因在洄游型刀鲚嗅囊中的表达量总体高于定居型,且洄游型雄性刀鲚嗅囊中此基因的表达量约是其雌性嗅囊中的3倍。这表明mor-4k13基因不仅与嗅觉功能和性腺发育相关,也可能与生殖洄游习性相关,不同性别的个体间也存在着嗅觉能力的差异。     

Differential development of odorant receptor expression patterns in the olfactory epithelium: a quantitative analysis in the mouse septal organ

The rodent olfactory epithelium expresses more than 1000 odorant receptors (ORs) with distinct patterns, yet it is unclear how such patterns are established during development. In the current study, we investigated development of the expression patterns of different ORs in the septal organ, a small patch of olfactory epithelium predominantly expressing nine identified ORs. The presumptive septal organ first appears at about embryonic day 16 (E16) and it completely separates from the main olfactory epithelium (MOE) at about postnatal day 7 (P7). Using in situ hybridization, we quantified the densities of the septal organ neurons labeled by specific RNA probes of the nine abundant OR genes from E16 to postnatal 3 months. The results indicate that olfactory sensory neurons (OSNs) expressing different ORs have asynchronous temporal onsets. For instance, MOR256-17 and MOR236-1 cells are present in the septal organ at E16; however, MOR0-2 cells do not appear until P0. In addition, OSNs expressing different ORs show distinct developmental courses and reach their maximum densities at different stages ranging from E16 (e.g. MOR256-17) to 1 month (e.g. MOR256-3 and MOR235-1). Furthermore, early onset does not correlate with high abundance in adult. This study reveals a dynamic composition of the OSNs expressing different ORs in the developing olfactory epithelium.     

Elevated CO2 impairs olfactory‐mediated neural and behavioral responses and gene expression in ocean‐phase coho salmon (Oncorhynchus kisutch)

Elevated concentrations of CO₂ in seawater can disrupt numerous sensory systems in marine fish. This is of particular concern for Pacific salmon because they rely on olfaction during all aspects of their life including during their homing migrations from the ocean back to their natal streams. We investigated the effects of elevated seawater CO₂ on coho salmon (Oncorhynchus kisutch) olfactory‐mediated behavior, neural signaling, and gene expression within the peripheral and central olfactory system. Ocean‐phase coho salmon were exposed to three levels of CO₂, ranging from those currently found in ambient marine water to projected future levels. Juvenile coho salmon exposed to elevated CO₂ levels for 2 weeks no longer avoided a skin extract odor that elicited avoidance responses in coho salmon maintained in ambient CO₂ seawater. Exposure to these elevated CO₂ levels did not alter odor signaling in the olfactory epithelium, but did induce significant changes in signaling within the olfactory bulb. RNA‐Seq analysis of olfactory tissues revealed extensive disruption in expression of genes involved in neuronal signaling within the olfactory bulb of salmon exposed to elevated CO₂, with lesser impacts on gene expression in the olfactory rosettes. The disruption in olfactory bulb gene pathways included genes associated with GABA signaling and maintenance of ion balance within bulbar neurons. Our results indicate that ocean‐phase coho salmon exposed to elevated CO₂ can experience significant behavioral impairments likely driven by alteration in higher‐order neural signal processing within the olfactory bulb. Our study demonstrates that anadromous fish such as salmon may share a sensitivity to rising CO₂ levels with obligate marine species suggesting a more wide‐scale ecological impact of ocean acidification.     

A new concept of olfactory biosensor based on interdigitated microelectrodes and immobilized yeasts expressing the human receptor OR17-40

This work shows the feasibility of an olfactory biosensor based on the immobilization of Saccharomyces cerevisiae yeast cells genetically modified to express the human olfactory receptor OR17-40 onto interdigitated microconductometric electrodes. This olfactory biosensor has been applied to the detection of its specific odorant (helional) with a high sensitivity (threshold 10⁻¹⁴ M). In contrast, no significant response was observed using a non-specific odorant (heptanal), which suggests a good selectivity. Thus, this work may represent a first step towards a new kind of bioelectronic noses based on whole yeast cells and allowing a real time monitoring of olfactory receptor activation. Presented at the joint biannual meeting of the SFB-GEIMM-GRIP, Anglet, France, 14–19 October, 2006.     

Expression pattern of olfactory receptor genes in human cumulus cells as an indicator for competent oocyte selection

Odorant or olfactory receptors are mainly localized in the olfactory epithelium for the perception of different odors. Interestingly, many ectopic olfactory receptors with low expression levels have recently been found in nonolfactory tissues to involve in local functions. Therefore, we investigated the probable role of the olfactory signaling pathway in the surrounding microenvironment of oocyte. This study included 22 women in intracytoplasmic sperm injection cycle. The expression of olfactory target molecules in cumulus cells surrounding the growing and mature oocytes was evaluated by Western blotting and real-time polymerase chain reaction. Additionally, integrated bioinformatics analyses were carried out and 6 ectopic olfactory receptors were selected for further evaluation. The initiation of olfactory transduction cascade in cumulus cells of competent oocytes was confirmed by analyzing the expression of adenylyl cyclase type 3 and olfactory market protein. Moreover, the expression pattern of the selected olfactory receptors was evaluated and OR10H2 was selected due to a high level of expression in mature fertile oocytes. We suggested that OR10H2 could be considered as a reliable biomarker for oocyte selection in assisted reproduction technique programs. However, further studies are required to elucidate the role of olfactory transduction cascade in embryo quality and implantation.