Synchrotron X-ray tomographic microscopy of fossil embryos

Fossilized embryos from the late Neoproterozoic and earliest Phanerozoic have caused much excitement because they preserve the earliest stages of embryology of animals that represent the initial diversification of metazoans. However, the potential of this material has not been fully realized because of reliance on traditional, non-destructive methods that allow analysis of exposed surfaces only, and destructive methods that preserve only a single two-dimensional view of the interior of the specimen. Here, we have applied synchrotron-radiation X-ray tomographic microscopy (SRXTM), obtaining complete three-dimensional recordings at submicrometre resolution. The embryos are preserved by early diagenetic impregnation and encrustation with calcium phosphate, and differences in X-ray attenuation provide information about the distribution of these two diagenetic phases. Three-dimensional visualization of blastomere arrangement and diagenetic cement in cleavage embryos resolves outstanding questions about their nature, including the identity of the columnar blastomeres. The anterior and posterior anatomy of embryos of the bilaterian worm-like Markuelia confirms its position as a scalidophoran, providing new insights into body-plan assembly among constituent phyla. The structure of the developing germ band in another bilaterian, Pseudooides, indicates a unique mode of germ-band development. SRXTM provides a method of non-invasive analysis that rivals the resolution achieved even by destructive methods, probing the very limits of fossilization and providing insight into embryology during the emergence of metazoan phyla.     

"寒武纪爆发"事件及其成因述评

主要后生动物门类化石在寒武纪初期的突然出现记录了生命史中一个从未有过的快速演化时期,这一事件被称为“寒武纪爆发(Cambrian explosion)”．多年来有关“寒武纪爆发”成因的各种猜测从未间断过．但是,将古生物化石记录看成是生物进化过程中种系形成的真实过程已遭到许多研究者的非议,这主要取决于以下二方面的证据：一是对早期后生动物系统发生关系的重新评估;二是来自于分子系统学和分子钟的研究．现在看来,“寒武纪爆发”事件的产生,一方面归因于化石体积的明显增加及其伴随的骨骼化过程,另一方面可能是其时的进化革新事件所导致．     

Tetraradial symmetry in early poriferans

Here is currently little consensus on the branching order and phyletic status of the oldest metazoan groups, but sponges are widely believed to be the earliest- branching living metazoans. Porifera are thought to have diverged before the emergence of developmental charac- ters typical of Eumetazoa, such as well-defined symmetry; extant sponges show radial symmetry of indeterminate high order, or none, combined with polarisation along the axis. In contrast, other early-branching phyla include bilateral and tetraradial （Cnidaria） and biradial （Cte- nophora） symmetry, or none （Placozoa）. A variety of prismatic early fossil sponges had shown here where the shared symmetry has been overlooked, and also describe structural tetraradial symmetry in Cambrian sponges from South China. Based on this study, this symmetry is likely to have been a primitive feature of sponges, and that the earliest-known fossil sponges were highly organised, cel- lularly integrated individuals whose body form was under strict genetic control.     

贵州新元古代瓮安生物群中动物胚胎化石研究进展

 瓮安生物群指产出于贵州瓮安磷矿地区新元古代埃迪卡拉纪陡山沱组（635~551Ma）上磷矿段的一个特殊磷酸盐化微体化石组合,主要由多细胞藻类、大型带刺疑源类和处于不同发育阶段的多种后生动物胚胎组成,并包括少量可能的后生动物幼虫和成体化石。瓮安生物群作为迄今为止世界上最古老的前寒武纪后生动物化石群,为探索后生动物的起源和早期演化历程提供了独一无二的实证记录。10余年来,瓮安生物群中动物胚胎化石的研究取得多项突破性进展,已确认至少5种不同卵裂方式的动物胚胎化石,胚胎化石分类从单纯的形态学分类向生物学分类转变,为揭示瓮安生物群中动物多样性提供了更多线索。同时,相关研究更新了繁盛于埃迪卡拉纪大型带刺疑源类生物属性的传统认识,并将之与有表面装饰的动物休眠卵（滞育胚胎）化石相联系。同步辐射X射线显微CT技术的应用成功解决了胚胎化石三维无损成像难题,为古胚胎学的发展带来了新的机遇。     

瓮安生物群中"内陷式动物原肠胚化石"的同步辐射无损成像及其亲缘关系的重新解释

 贵州瓮安埃迪卡拉纪陡山沱组(635—551Ma)的磷酸盐化动物胚胎化石作为全球已知最古老的多细胞动物化石记录,为研究后生动物的起源及早期演化过程提供了精美的实证材料.在传统扫描电子显微成像的基础上,结合同步辐射相位衬度显微断层成像技术,对瓮安生物群中以前报道的"内陷式动物原肠胚化石"的立体标本进行了比较形态学和解剖学研究.结果显示,贵州瓮安陡山沱组磷酸盐化内陷式动物原肠胚化石很可能是多细胞藻类,并非动物胚胎.     

Early blastomere determines embryo proliferation and caste fate in a polyembryonic wasp

Polyembryonic development is a unique mode of metazoan development in which a single zygote generates multiple embryos by clonal proliferation. The polyembryonic parasitic insect Copidosoma floridanum shows one of the most extreme cases of polyembryony, producing up to 2,000 embryos from a single egg. In addition, this wasp exhibits an unusual polyphenism, producing two morphologically distinct larval castes, termed precocious and reproductive, that develop clonally from the same zygote. This form of development seems incompatible with a model of insect development in which maternal pre-patterning of the egg specifies embryonic axial polarity. Here we show that maternal pre-patterning in the form of germ plasm creates cellular asymmetry at the four-cell stage embryo of Copidosoma that is perpetuated throughout development. Laser ablations of cells show that the cell inheriting the germ plasm regulates both the fate and proliferation of the reproductive caste. Thus, we have uncovered a new mechanism of caste specification, mediated by the regulatory capacity of a single cell. This study shows that the evolution of mammalian-like regulative development of an insect embryo relies on a novel cellular context that might ultimately enhance developmental plasticity.     

Determination of spatial domains of zygotic gene expression in the Drosophila embryo by the affinity of binding sites for the bicoid morphogen

The maternal gene bicoid is a key component of the system that determines the pattern of the anterior half of Drosophila embryos. The bicoid protein forms a concentration gradient in early embryos, and is known to bind DNA. Specific binding sites are now shown to confer expression in a region of the embryo that depends on their affinity for bicoid protein: sites of high affinity allow expression further down the bicoid protein gradient than sites of low affinity.     

An epipodite-bearing crown-group crustacean from the Lower Cambrian

Crown-group crustaceans (Eucrustacea) are common in the fossil record of the past 500 million years back to the early Ordovician period, and very rare representatives are also known from the late Middle and Late Cambrian periods. Finds in Lower Cambrian rocks of the Phosphatocopina, the fossil sister group to eucrustaceans, imply that members of the eucrustacean stem lineage co-occurred, but it remained unclear whether crown-group members were also present at that time. 'Orsten'-type fossils are typically tiny embryos and cuticle-bearing animals, of which the cuticle is phosphatized and the material is three-dimensional and complete with soft parts. Such fossils are found predominantly in the Cambrian and Ordovician and provide detailed morphological and phylogenetic information on the early evolution of metazoans. Here we report an Orsten-type Konservat-Lagerst?tte from the Lower Cambrian of China that contains at least three new arthropod species, of which we describe the most abundant form on the basis of exceptionally well preserved material of several growth stages. The limb morphology and other details of this new species are markedly similar to those of living cephalocarids, branchiopods and copepods and it is assigned to the Eucrustacea, thus representing the first undoubted crown-group crustacean from the early Cambrian. Its stratigraphical position provides substantial support to the proposition that the main cladogenic event that gave rise to the Arthropoda was before the Cambrian. Small leaf-shaped structures on the outer limb base of the new species provide evidence on the long-debated issue of the origin of epipodites: they occur in a set of three, derive from setae and are a ground-pattern feature of Eucrustacea.     

Developmental sequence of Cambrian embryo Markuelia

Based on more exquisitely preserved specimens of Markuelia hunanensis recently recovered from Middle and Upper Cambrian in western Hunan and in the light of Synchrotron radiation X-ray tomo-graphic microscopy, the developmental sequence from cleavage through organogenesis to the pre-hatching of Cambrian embryo Markuelia, especially the developmental sequence during the pre-hatching stage, i.e. from the earliest period when the scalids and tail spines only took shape to the latest period (just about hatching), is established. This developmental sequence provides a pattern of embryonic development during the pre-hatching stage, which has not been established in the living scalidophorans (priapulids, loriciferans and kinorhynchs). Thus, it not only enriches our knowledge on the embryonic development of the extant descendants of Markuelia, but also opens a new window to the evolution and development of the animal.     

Embryonic and extraembryonic stem cell lines derived from single mouse blastomeres

The most basic objection to human embryonic stem (ES) cell research is rooted in the fact that ES cell derivation deprives embryos of any further potential to develop into a complete human being. ES cell lines are conventionally isolated from the inner cell mass of blastocysts and, in a few instances, from cleavage stage embryos. So far, there have been no reports in the literature of stem cell lines derived using an approach that does not require embryo destruction. Here we report an alternative method of establishing ES cell lines-using a technique of single-cell embryo biopsy similar to that used in pre-implantation genetic diagnosis of genetic defects-that does not interfere with the developmental potential of embryos. Five putative ES and seven trophoblast stem (TS) cell lines were produced from single blastomeres, which maintained normal karyotype and markers of pluripotency or TS cells for up to more than 50 passages. The ES cells differentiated into derivatives of all three germ layers in vitro and in teratomas, and showed germ line transmission. Single-blastomere-biopsied embryos developed to term without a reduction in their developmental capacity. The ability to generate human ES cells without the destruction of ex utero embryos would reduce or eliminate the ethical concerns of many.     

Derivation of androgenetic embryonic stem cells from m-carboxycinnamic acid bishydroxamide (CBHA) treated androgenetic embryos

The androgenetic embyronic stem (aES) cells are useful models in studying the effects of imprinted genes on pluripotency maintaining and embryo development. The expression patterns of imprinted genes are significantly different between uniparental derived aES cells and zygote-derived embryonic stem (ES) cells, therefore, the imprinting related cell pluripotency needs further exploitation. Several approaches have been applied in generation of androgenetic embryos and derivation of aES cell lines. Here, we describe a method to generate androgenetic embryos by injecting two mature sperms into one enucleated oocyte. Then these androgenetic embryos were treated with a histone deacetylase inhibitor: m-carboxycinnamic acid bishydroxamide (CBHA). Further, aES cell lines were successfully derived from these treated androgenetic embryos at blastocyst stage. The CBHA could improve not only the quality of androgenetic embryos, but also the efficiencies of aES (CaES) cells derivation and chimeric mice generation. The imprinted gene expression pattern in the CBHA treated embryo-derived aES (CaES) cells was also highly similar to that of zygote-derived ES cells.     

Oceanic 18S rDNA sequences from picoplankton reveal unsuspected eukaryotic diversity

Picoplankton--cells with a diameter of less than 3 microm--are the dominant contributors to both primary production and biomass in open oceanic regions. However, compared with the prokaryotes, the eukaryotic component of picoplankton is still poorly known. Recent discoveries of new eukaryotic algal taxa based on picoplankton cultures suggest the existence of many undiscovered taxa. Conventional approaches based on phenotypic criteria have limitations in depicting picoplankton composition due to their tiny size and lack of distinctive taxonomic characters. Here we analyse, using an approach that has been very successful for prokaryotes but has so far seldom been applied to eukaryotes, 35 full sequences of the small-subunit (18S) ribosomal RNA gene derived from a picoplanktonic assemblage collected at a depth of 75 m in the equatorial Pacific Ocean, and show that there is a high diversity of picoeukaryotes. Most of the sequences were previously unknown but could still be assigned to important marine phyla including prasinophytes, haptophytes, dinoflagellates, stramenopiles, choanoflagellates and acantharians. We also found a novel lineage, closely related to dinoflagellates and not previously described.     

鹅掌楸属树种的胚胎学研究进展

目前,鹅掌楸属树种的胚胎学研究可以划分为3 个阶段,即描述胚胎学阶段、比较胚胎学阶段以及实验胚胎学阶段,研究内容涉及了雌、雄配子体的构建,传粉与受精作用的完成,胚与胚乳的发育及种子的形成,性细胞的分离以及未成熟胚的培养等方面。笔者概述了鹅掌楸属树种的一般胚胎学过程与杂交胚胎学的研究结果,以及实验胚胎学的研究进展,并针对鹅掌楸属树种生殖细胞工程和体细胞胚胎发生系统的建立等研究领域进行了展望性的讨论。     

An armoured Cambrian lobopodian from China with arthropod-like appendages

Cambrian fossil Lagerst?tten preserving soft-bodied organisms have contributed much towards our understanding of metazoan origins. Lobopodians are a particularly interesting group that diversified and flourished in the Cambrian seas. Resembling 'worms with legs', they have long attracted much attention in that they may have given rise to both Onychophora (velvet worms) and Tardigrada (water bears), as well as to arthropods in general. Here we describe Diania cactiformis gen. et sp. nov. as an 'armoured' lobopodian from the Chengjiang fossil Lagerst?tte (Cambrian Stage 3), Yunnan, southwestern China. Although sharing features with other typical lobopodians, it is remarkable for possessing robust and probably sclerotized appendages, with what appear to be articulated elements. In terms of limb morphology it is therefore closer to the arthropod condition, to our knowledge, than any lobopodian recorded until now. Phylogenetic analysis recovers it in a derived position, close to Arthropoda; thus, it seems to belong to a grade of organization close to the point of becoming a true arthropod. Further, D. cactiformis could imply that arthropodization (sclerotization of the limbs) preceded arthrodization (sclerotization of the body). Comparing our fossils with other lobopodian appendage morphologies--see Kerygmachela, Jianshanopodia and Megadictyon--reinforces the hypothesis that the group as a whole is paraphyletic, with different taxa expressing different grades of arthropodization.     

Lower Cambrian yolk-pyramid embryos from Southern Shaanxi, China

TheCambrianexplosioniswidelyacceptedasthesuddenappearanceofnumerousbilateriananimalphylaatornearthebeginningofCambriantime[1,2 ] .The 5 30 million year oldMaotianshanShalefaunacontainstheoldestgoodwhole bodyfossilsofbilateri ans,documentinganincreasingnumberofpresent dayanimalphyla[3,4 ] orsubphyla[5] (evenincludingvertebrates[6～ 8] aswell)knownfromLowerCambri an .ThebeginningoftheCambrianperiodisdatedat5 43millionyearsago ,whenthefirstlargeandelabo ratefossilburrowstogetherwiththemicrosco…     

Human pregnancy following cryopreservation, thawing and transfer of an eight-cell embryo

The widespread use of clomiphene citrate and exogenous gonadotrophins for in vitro fertilization (IVF) in human frequently results in the production of multiple embryos. Replacement of more than two embryos increases pregnancy rate but may result in multiple pregnancies with increased pre- and post-natal abnormality. Preservation of embryos for a limited time allows fewer embryos to be replaced on several different occasions and thus the problems of multiple pregnancy can be minimized, the effectiveness of a single IVF procedure increased and embryo replacement in adverse maternal conditions avoided. Preimplantation embryos have been successfully cryopreserved in many animal species. The sensitivity of embryos to cooling and freezing varies between species and stages of embryo development. We report here the cryopreservation procedures that allow a high survival rate of four- and eight-cell human embryos and the establishment of a pregnancy following the freezing and storage of an eight-cell embryo for 4 months in liquid nitrogen. The pregnancy terminated at 24 weeks' gestation due to development of a septic Streptomyces agalactiae chorion amnionitis after premature membrane rupture.     

Discovery of phosphatized gastrula fossils from the Doushantuo Formation, Weng’an, Guizhou Province, China

Globular fossils from Doushantuo phosphorites at the Weng’an area are for the first time identified as a gastrulation stage of phosphatized embryos. They are got from the fossiliferous remains after acetic acid maceration. The fossils are found together with formerly reported animal resting eggs and embryos of the earlier cleavage stage. The oblate-shaped fossils with the same size as those reported embryos and invaginate at the middle part into the embryos, show the characteristics of the late blastula to the early gastrula stage of the embryo development. This discovery convinces the existence of animal embryos at Doushantuo age and offers new facts for the studying of the affinity of related fossils, which are still controversial at present.     

后生动物起源与进化的分子系统学研究进展

后生动物的起源与进化问题一直是动物学、古生物学和进化生物学研究的重要课题,传统上,对其系统发生和亲缘关系的探讨主要是基于比较形态学和比较胚胎学的证据,然而,随着分子遗传学和分子发育生物学的不断进展及其在相关研究领域的普遍运用,从遗传基础和发育机理上追踪动物各分类群间的系统发生关系已经成为可能。本文就近年来后生动物高级分类群（主要为“门”一级）的分子系统发生研究及其研究中存在的一些问题作了较为系统的综述,同时就后生动物系统发生研究的前景作了展望。     

小鼠着床前胚胎Dicer 1和DGCR 8基因的表达

研究了miRNA生成相关基因Dicer 1和 DGCR 8在小鼠着床前胚胎各发育阶段的表达,探讨其对着床前胚胎发育的作用.建立小鼠超排、交配系统,采集着床前胚胎;采用二步法RT-PcR方法鉴定着床前胚胎Dicer1和DGcR 8表达.结果显示:小鼠卵母细胞及受精后1.5 d、2.5 d、3.5 d和4.5 d着床前胚胎均表达Dicef 1和DGcR 8基因.这是着床前胚胎IlliRNA生成物的重要基因,miRNA及其生物合成相关基因可能对着床前胚胎发育起重要作用.