Zebrafish as the Best Vertebrate Model for Development Studies

从八十年代初开始的、通过突变体分离早期胚胎发育基因的研究,揭示了无脊椎动物果蝇形态发生的分子机制,成为１９９５年诺贝尔生理或医学奖的主要内容,同时也标志着发育生物学已经成为生物学的带头学科。该成果的取得得益于果蝇具备发育生物学研究模型的特点,即可同时进行胚胎学和发育遗传学研究。在脊椎动物发育生物学研究方面,由于缺少适当的研究模型,有关早期胚胎发育基因表达调控的研究始终未获突破性进展。包括小鼠、鸡、爪蟾等在内的一些脊椎动物都只适用于胚胎学或者遗传学某一学科的研究,均不是理想的发育生物学研究模型。一种小型的鲤科鱼———斑马鱼,逐渐成为发育生物学研究的最佳脊椎动物模型。它光周期产卵,卵大,胚胎在体外发育,胚胎发育速度快,早期胚胎完全透明,这些特点使它成为很好的脊椎动物胚胎学研究模型;同时,它个体小,产卵量大,产卵周期短,单倍体、雌核发育二倍体的制作和突变体的获得均较容易,精子可以冷冻保存,所有这些特点又使斑马鱼非常适合于发育遗传学研究。本文将详细论述斑马鱼作为脊椎动物发育生物学研究模型的特点,以及作为模型动物正在进行的有关工作。     

非洲爪蟾体节发生标志基因Mespo的表达调控研究

脊椎动物的脊柱来源于体节,体节是胚胎发生过程中的过渡性结构。体节的形成过程被称为体节发生（somitogenesis）。胚胎发育过程中正确的分节保证了脊椎动物具有正常的体轴,因此体节发生在时间和空间上受到严格的遗传控制,对体节发生的分子机制的研究是发育生物学中的重要课题。     

两侧对称动物左右不对称发生机制研究进展

左右不对称是两侧对称动物的重要特征,其形成机制一直是发育生物学领域备受关注的科学问题之一。脊椎动物的左右不对称发生经过3个重要阶段：左右对称性的打破,左右不对称信号的建立和维持,以及左右不对称器官的形态发生。多数脊椎动物在胚胎发育阶段依赖纤毛产生定向液流打破胚胎的左右对称性,随后建立Nodal-Pitx2左右不对称信号,最后由Pitx2等基因指导左右不对称器官的形态发生过程。无脊椎动物中存在不依赖纤毛介导的Nodal-Pitx不对称信号表达机制,甚至具有完全独立的左右不对称发育机制。本文结合最新的左右不对称器官发育机制的研究进展,综述了脊椎动物和无脊椎动物胚胎左右不对称的发生过程及相关基因和信号通路,有助于深入理解左右不对称器官发育的过程,以期为追溯左右不对称器官发育机制的起源演化提供参考。     

斑马鱼外胚层早期发育及其某些调控机制

斑马鱼已经成为脊椎动物发育生物学研究的一种常用模式生物。它的体形较小,价格低廉,受精和发育都在体外,胚胎的通体透明,可以用活体观察基因突变后引起的表型变化。该阐明斑马鱼外胚层的发育及其某些调控机制。     

斑马鱼—发育生物学研究的最佳脊椎动物模型

从八十年代初开始的、通过突变体分离早期胚胎发育基因的研究,揭示了无脊椎动物果蝇形态发生的分子机制,成为1995年诺贝尔生理或医学奖的主要内容,同时也标着发育生物学已经成为生物学的带头学科。该成果的取得得益于果蝇具备发育生物学研究模型的特点,即可同时进行胚胎学和发育遗传学研究。在脊动物发育生物学研究方面,由于缺少适当的研究模型,有关早期胚胎发育基因表达调鬼斧神工的研究始终未获突破性进展。包括小鼠、鸡、爪蟾等在一些脊椎动物都只适用于胚胎学或者遗传学某一学科的研究,均不是理想的发育生物学研究模型。一种小型的鲤科鱼－斑马鱼,逐渐成为发育生物学研究的最佳脊椎运动模型。它光周期产卵、卵大,胚胎在体外发育,胚胎发育速度快,早期胚胎完全透明,这些特点使它成为很好的脊椎动物胚胎学研究模型;同时,它个体小,产卵量大,产卵周期短,单倍体、雌核发育二倍体的制作和突变体的获得均较容易,精子可以冷冻保存,所有这些特点又使斑马鱼非常适合于发育遗传学研究。本文将详细论述斑马鱼作为脊椎动物发育生物学研究模型的特点,以及作为模型动物正在进行的有关工作。     

亚·奥·柯瓦列夫斯基与脊索动物

亚历山大·奥努夫里耶维奇·柯瓦列夫斯基(1840—1901),是十九世纪俄国生物学家,进化论比较胚胎学的开创人。他和梅契尼科夫合作,采用比较的方法,在世界上首次广泛研究无脊椎动物的胚胎发育。证明无脊椎动物与脊椎动物都有相似的胚层,各胚层发育而成的组织、器官也基本相似,建立了胚层学说,肯定了无脊椎动物与脊椎动物的亲缘关系。他还对多种脊索动物的个体发育进行了精深的研究。关于文昌鱼研究的科学记载也是他在《蛞蝓鱼的发育》一文中,首先指出了文昌鱼发育过程中的双重性。在胚胎发育上,像脊椎动物的胚胎通过相似阶段而发育;但在早期又…     

斑马鱼早期内胚层发育及其分子调控机制

斑马鱼已成为研究脊椎动物胚胎发育的理想模式生物,它具有体外受精、发育周期快、胚体透明等优点,此外其胚胎发育过程中的信号通路与哺乳动物有很高的同源性。该文阐述了斑马鱼早期内胚层发育的过程及其分子调控机制。     

海洋无脊椎动物甲状腺激素信号通路的研究进展

在脊椎动物中,甲状腺激素信号通路是调控生长、发育和机体能量代谢必不可少的信号通路之一,并且参与了两栖类和鱼类的变态反应。近来,越来越多的证据表明,在海洋无脊椎动物中存在内源性的甲状腺激素、甲状腺激素受体等信号通路的成员分子,而且这些分子参与了海洋无脊椎动物的发育和变态过程。这表明在海洋无脊椎动物中存在与脊椎动物类似的甲状腺激素信号通路。综述了海洋无脊椎动物中甲状腺激素信号通路的相关研究进展,旨在为研究甲状腺激素在海洋无脊椎动物的生物学功能及其作用机制提供基础资料。     

文昌鱼左右体轴建立机制的研究进展

两侧对称动物左右体轴建立机制研究是发育生物学领域重要的基础科学问题之一.文昌鱼(amphioxus)由于其特殊的进化地位以及与脊椎动物相似的胚胎发育模式和身体构筑方式,是研究动物左右体轴建立机制的理想模式物种.近年来随着文昌鱼室内全人工繁育技术、高效显微注射技术和基因敲除技术的建立,国内外学者在左右体轴建立机制研究上取...     

Hox基因在中枢神经系统中作用的研究进展

同源异型盒基因(homeobox genes,Hox)在进化上是保守的,在调节胚胎发育,尤其在脊椎动物前–后轴(antero-posterior axis)的发育及肢体发育中起着重要作用。最近研究表明,Hox基因通过与之相关的辅助因子调节中枢神经系统(central nervous system,CNS)以及神经环路的发育。该文主要对Hox基因调控中枢神经系统发育的作用及机制进行综述。     

ycaCR基因在文昌鱼早期胚胎表达研究

文昌鱼作为现存的与脊椎动物最接近的无脊椎动物,一直被作为研究生物进化和胚胎发育的典型材料.利用整体原位杂交方法对从文昌鱼肠cDNA文库克隆到的ycaCR基因进行基因的胚胎表达模式研究,结果显示该基因在早期胚胎发育阶段没有表达,在2天幼虫的原始消化道表达,暗示ycaCR基因可能在原始消化道内发挥生物学作用.     

Characterization and expression of p23 gene in the amphioxus Branchiostoma belcheri

The cDNA AmphiP23, encoding an amphioxus p23, was identified from the gut cDNA library of amphioxus Branchiostoma belcheri. It contains a 513 bp open reading frame corresponding to a deduced protein of 170 amino acids. Phylogenetic analysis shows that vertebrate and invertebrate p23/p23-like proteins are each grouped together, with AmphiP23 falling at the base of vertebrate p23/p23-like clade, suggesting that the divergence of vertebrate and invertebrate p23 genes probably occurs prior to the split of invertebrate/vertebrate from a common ancestor around 550 million years ago. Northern blotting reveals a ubiquitous expression pattern of AmphiP23 in all adult tissues examined, while whole mount in situ hybridization demonstrates a tissue- and stage-specific expression pattern of AmphiP23 in developing embryos and larvae. Presumably, the ubiquitous expression pattern of AmphiP23 in adult amphioxus represents the ancestral type of p23 gene prior to its split to human paralogs p23 and tsp23, while the tissue- and stage-specific expression pattern during early embryonic development implicates a role of AmphiP23 in anterior/posterior patterning.     

文昌鱼AmphiRab23b基因的克隆、进化分析及表达模式

Hedgehog信号通路在胚胎发育过程中发挥着重要作用, 同时与多种肿瘤的发生密切相关。Rab23蛋白在Hedgehog信号通路中扮演着十分重要的角色。目前关于文昌鱼Rab23同源基因的研究仅局限于佛罗里达文昌鱼(Branchiostoma floridae)基因组中的注释。文章首次克隆了中国文昌鱼(Branchiostoma belcheri) Rab23b基因 (AmphiRab23b)cDNA全序列 , 对其演译的蛋白序列进行了序列比对、进化树分析以及基因时空表达分析。研究结果显示, 文昌鱼AmphiRab23b基因的 cDNA总长为2 062 bp (包括UTR区), 其中开放阅读框 (Open reading frame, ORF) 714 bp, 编码237个氨基酸; 虽然在进化树中不属于脊椎动物Rab23进化支, 但是AmphiRab23具有保守的Rab23_lke结构域, 暗示该基因在进化过程中可能在功能上是保守的。时空表达的研究结果进一步显示, AmphiRab23b基因在胚胎发育中的神经板和消化道中表达, 与其脊椎动物同源基因的表达模式相似, 这说明该基因可能对文昌鱼神经系统和消化道的发育有重要作用。     

Developmental expression of amphioxus GABA<Subscript>A</Subscript> receptor-associated protein-like 2 gene

A full-length amphioxus -aminobutyric acid type-A receptor-associated protein-like 2 (GABARAPL2) cDNA was isolated. Its sequence and developmental expression are first described in this paper. The phylogenetic analysis shows that the amphioxus GABARAPL2 and GABARAPL2 in vertebrates are highly homologous. The results of in situ hybridization show that the amphioxus GABARAPL2 gene is expressed in the neural tube, neurenteric canal, notochord, muscle and developing alimentary canal.Edited by P. SimpsonKailong Liang and Yushuang Lin contributed equally to the study     

Characterization and expression of amphioxus ApoD gene encoding an archetype of vertebrate ApoD proteins

Here we report a homologue of the apolipoprotein D gene (AmphiApoD) in amphioxus, Branchiostoma belcheri tsingtauense, the first such finding in a basal chordate cephalochordate. The main features of the protein predicted from AmphiApoD are characteristic of the apolipoprotein D. Phylogenetic analysis places AmphiApoD at the base of the phylogenetic tree, suggesting that AmphiApoD is the archetype of the vertebrate ApoD genes. Both whole mount in situ hybridization and Northern blotting and RT-PCR as well as in situ hybridization histochemistry reveal that AmphiApoD is expressed in tissues derived from mesoderm and endoderm including notochord and hind-gut, which contrasts with the strong expression patterns of ApoD genes in the ectodermal derivatives in mammals and birds. The expression profiles of the ApoD gene may have been changed to be expressed in the endo-mesodermal derivatives in amphioxus after the vertebrate and cephalochordate lineages diverged; alternatively, the ApoD gene may first have been expressed in the endo-mesoderm during embryogenesis in the last common ancestor of all chordates, and subsequently came to be expressed in the ectodermal derivatives of vertebrates including mammals and birds.     

Retinoic acid signaling acts via Hox1 to establish the posterior limit of the pharynx in the chordate amphioxus

In the invertebrate chordate amphioxus, as in vertebrates, retinoic acid (RA) specifies position along the anterior/posterior axis with elevated RA signaling in the middle third of the endoderm setting the posterior limit of the pharynx. Here we show that AmphiHox1 is also expressed in the middle third of the developing amphioxus endoderm and is activated by RA signaling. Knockdown of AmphiHox1 function with an antisense morpholino oligonucleotide shows that AmphiHox1 mediates the role of RA signaling in setting the posterior limit of the pharynx by repressing expression of pharyngeal markers in the posterior foregut/midgut endoderm. The spatiotemporal expression of these endodermal genes in embryos treated with RA or the RA antagonist BMS009 indicates that Pax1/9, Pitx and Notch are probably more upstream than Otx and Nodal in the hierarchy of genes repressed by RA signaling. This work highlights the potential of amphioxus, a genomically simple, vertebrate-like invertebrate chordate, as a paradigm for understanding gene hierarchies similar to the more complex ones of vertebrates.     

光棘球海胆seali基因克隆及表达特性分析

seali基因隶属于PIWI超家族, 其编码的RNA结合蛋白在生殖细胞发育过程中发挥重要作用。研究经同源比对从光棘球海胆(Mesocentrotus nudus)性腺转录组数据库中筛选得到seali基因片段, 随后通过cDNA末端快速扩增技术(Rapid amplification of cDNA ends, RACE), 获得其全长cDNA序列。光棘球海胆seali基因cDNA全长3462 bp, 其中3′UTR长度为416 bp, 5′UTR长度为180 bp, 其中3′UTR的加尾信号并非经典的AAUAAA或AUUAAA, 而是较少见的AAUACA。开放阅读框(Open Reading Frame, ORF)2862 bp, 编码954个氨基酸, 具有保守的PIWI和PAZ结构域, 多重序列比对和系统进化分析结果表明其属于Argonaute家族的PIWI亚家族成员。荧光定量PCR技术检测结果表明, Mnseali基因在光棘球海胆性腺、肠、管足和体腔细胞中均有表达, 在性腺中表达量最高。此外, Mnseali基因为母源因子, 在整个胚胎发育时期均有表达。在卵巢中, 随着卵母细胞的成熟, Mnseali的表达量逐渐升高, 而仅在成熟期的精巢中表达量显著上调。RNA原位杂交结果表明, Mnseali在光棘球海胆性腺的生殖细胞中特异表达, 是光棘球海胆生殖细胞标记基因。该研究为海胆生殖细胞发育相关的研究提供了支撑。     

A trypsin homolog in amphioxus: expression, enzymatic activity and evolution

Trypsin has been documented in a variety of species including both vertebrates and invertebrates, but little is known about it in amphioxus, a model organism for insights into the origin and evolution of vertebrates. Here we identified a trypsin gene in Branchiostoma japonicum. The cDNA was 978 bp long with an ORF encoding a deduced protein of 272 amino acids. The deduced protein had an N-terminal signal peptide of 15 amino acids, a 16 activation peptide with the typical cleavage site Arg/Ile, a Tryp_SPc domain with the catalytic triad His⁷²-Asp¹¹⁸-Ser²¹⁵ and the S1 substrate binding residue Asp²⁰⁹, which are all characteristic of trypsinogens. The recombinant trypsin protein was able to hydrolyse the trypsin prototypic substrate BAEE, which was inhibited by the trypsin-specific inhibitor soybean trypsin inhibitor. Both northern blotting and tissue-section in situ hybridization demonstrated that trypsin gene was expressed in a tissue-specific manner, with most abundant levels in the hepatic caecum, mid-gut and ovary. And the whole mount in situ hybridization showed that it began to express in the middle third of the full-length primitive gut in 2-day larvae, where the hepatic caecum will form later during development. Phylogenetic analysis indicated that both amphioxus and ascidian trypsins are more closer to each other than to vertebrate trypsins, suggesting a continuous evolutionary divergence of vertebrate trypsins after split from protochordate/vertebrate common ancestor.