hUVECs在不同切应力作用下的形态及纤毛发生

分别应用紫杉醇荧光和免疫荧光标记法显示h UVECs(human umbilical vein endothelial cells,h UVECs)在不同切应力加载下微管骨架的装配动态及初级纤毛的形态发生。结果表明,以梯度切应力加载24 h,随着切应力增大,细胞由梭形逐渐变圆,长宽比降低,胞质微管向细胞核周围集结并发生装配;14 dynes/cm2切应力加载48 h后,微管向切应力方向延伸导致细胞呈长梭型,微管停止装配;以15 dynes/cm2切应力加载28 h后,纤毛基体在细胞表面的定位不可见,纤毛微管解聚;再以1 dynes/cm2切应力加载18 h,又可观察到纤毛基体在细胞表面的重新定位。因此,切应力作用可诱导h UVECs形态变化,该变化经历了微管骨架的解聚、向细胞核方向聚集及重新装配过程;此外,切应力能够影响初级纤毛的形态发生,该现象可能导致了初级纤毛在心血管系统中的分布不均,并为"纤毛疾病"的治疗提供新的思路。

初级纤毛发生起始的分子机制

初级纤毛是一种位于细胞表面基于微管的膜突起,在真核生物中广泛存在。作为细胞的“天线”,初级纤毛参与多种重要的细胞内信号转导通路,在动物组织器官的发育、分化及稳态维持中发挥重要作用。因此,纤毛结构和功能的缺陷会导致多种器官病变,引发一系列人类遗传疾病。纤毛由中心粒顶端延伸而出,但纤毛形成的起始过程目前尚不十分清楚。纤毛的发生起始涉及到中心粒/基体的锚定、中心粒帽蛋白CP110的去除以及纤毛芽的形成。该文将对近年来这三个方面的研究进展进行综述,为深入理解纤毛发生起始的分子机制提供参考。

A General Overview of the Typical 18 Frontal-VentralTransverse Cirri Oxytrichidae s. l. Genera(Ciliophora, Hypotrichia)

Oxytrichidae s. l. ciliates usually have 18 frontal-ventral-transverse cirri which are clustered to six distinct groups usu- ally originating from six longitudinal primordia segregating 1, 3, 3, 3, 4, 4 cirri. During morphogenesis, three dorsal kinetics anlagen are primarily formed. Fragmentation of kinety 3 usually present, while sometimes secondarily lost. Dorsomarginal kineties are formed, while sometimes lost. Oxytrichids tend to have overlapping characters, e.g. cell shape and size, infraciliature, pellicle fea- tures. This makes a great problem for genera separation. In the present work, all typical 18 frontal-ventral-transverse-cirri Oxytrichi- dae s. 1. genera were revised systematically based on their living morphology, ciliature patterns and dorsal morphogenetic features. The outline of the genera, the schematic illustrations, and the key to typical 18 frontal-ventral-transverse-cirri genera of Oxytrichidae s. l. were clarified. Additionally, some morphological and morphogenetic patterns were summarized and compared.

Phylogeny of subclass Scuticociliatia(Protozoa,Ciliophora) using combined data inferred from genetic,morphological,and morphogenetic evidence

Gene sequence-based genealogies of scuticociliates are different from those produced by morphological analyses.For this reason,11 representative scuticociliates and two ambiguously related genera were chosen to test the ability of combined phylogenetic analyses using both gene sequences and morphological/morphogenetic characteristics.Analyses of both the SSrRNA gene sequences and the combined datasets revealed a consistent branching pattern.While the terminal branches and the order level relationships were generally well resolved,the family level relationships remain unresolved.However,two other trees based on ITS1-5.8S-ITS2 region sequences and morphological/morphogenetic characters showed limited information,due to a lack of informative sites in these two datasets.Our data suggest,however,that the combined analysis of morphological/morphogenetic characters and gene sequences did produce some changes to the phylogenetic estimates of this group.

Insights into the phylogeny of sporadotrichid ciliates (Protozoa, Ciliophora: Hypotricha) based on genealogical analyses of multiple molecular markers

The sporadotrichid ciliates are an especially diverse group. A number of investigators have studied the morphological, morphogenetic, and molecular relationships among members of this group. Despite this, a consistent classification is still lacking and several important questions about the phylogenetic relationships within this group remain unsolved. To improve our understanding of these relationships, we constructed phylogenetic trees using the nucleotide sequences of the small-subunit rRNA (SSrRNA) gene and amino acid sequences of actin I and α-tubulin. Analyses of SSrRNA gene sequences indicated that: 1) the Sporadotrichida sensu Lynn (2008) and the Oxytrichidae are polyphyletic; 2) the Uroleptus species, which are classified to urostylids, formed a sister group with the oxytrichids; 3) Halteria grandinella, which is grouped morphologically with oligotrich species, clustered within the oxytrichids. These results are congruent with previous studies based on SSrRNA gene sequences. However, the amino acid sequences of actin I and α-tubulin yielded different topologies. The main results are: 1) in all phylogenetic trees, the genus Oxytricha was paraphyletic; 2) Uroleptus was sister to a subset of Urostyla and Holosticha, albeit with low supporting values; 3) Halteria grandinella was separated distantly from the Oxytrichidae in trees inferred from actin I amino acid sequences but clustered with oligotrichids in the α-tubulin analysis. The inconsistency among the trees inferred from these different molecular markers may be caused by rapidly accumulated genetic characterizations of ciliates. Further studies with additional molecular markers and sampling of more taxa are expected to better address the relationships among sporadotrichids.

Molecular Phylogenetic Lineage of Plagiopogon and Askenasia(Protozoa, Ciliophora) Revealed by Their Gene Sequences

Prostomates and haptorians are two basal groups of ciliates with limited morphological characteristics available for tax- onomy. Morphologically, the structures used to identify prostomates and haptorians are similar or even identical, which generate heavy taxonomic and phylogenetic confusion. In present work, phylogenetic positions lineage of two rare genera, Plagiopogon and Askenasia, were investigated. Three genes including small subunit ribosomal RNA gene （hereafter SSU rDNA）, internal transcribed spacer region （ITS region）, and large subunit ribosomal RNA gene （LSU rDNA） were analyzed, 10 new sequences five species each. Our findings included 1） class Prostomatea and order Haptorida are multiphyletic; 2） it may not be appropriate to place order Cyclotriehiida in subclass Haptoria, and the systematic lineage of order Cyclotrichiida needs to be verified further; 3） genus Plagio- pogon branches consistently within a clade covering most prostomes and is basal of clade Colepidae, implying its close lineage to Prostomatea; and 4） Askenasia is phylogenetically distant from the subclass Haptoria but close to classes Prostomatea, Plagiopylea and Oligohymenophorea. We supposed that the toxicyst of Askenasia may be close to taxa of prostomes instead of haptorians, and the dorsal brush is a more typical morphological characteristics of haptorians than toxicysts.

Evolution of discocephalid ciliates:Molecular,morphological and ontogenetic data support a sister group of discocephalids and pseudoamphisiellids (Protozoa,Ciliophora) with establishment of a new suborder Pseudoamphisiellina subord.n.

Discocephalids and pseudoamphisiellids are possibly two of the most confused groups among hypotrichous/euplotid ciliates regarding their systematic position and phylogenetic relationships.The former were often regarded as related to euplotids while the latter,in the absence of molecular data,were mostly assigned to the urostylid-like hypotrichs.In the present work,the small subunit rRNA genes of several rarely observed discocephalid and pseudoamphisiellid genera were analyzed to obtain insights into the phylogenetic relationships of these highly ambiguous Spirotrichea.Four different tree reconstruction algorithms yielded nearly identical topologies,which indicated both groups belong to the same assemblage.This assemblage is clearly isolated as a deep-branching clade and invariably positioned between Euplotida and Hypotricha.The sister group relationship of the Pseudoamphisiellidae and Discocephalidae supports the previous suggestion that they might represent an ordinal taxon,the Discocephalida.Both morphological and morphogenetic features indicate that the pseudoamphisiellids should be placed in the order Discocephalida but as a sister group to other typical discocephalids.Thus we propose establishing a new suborder,Pseudoamphisiellina subord.n.The new taxon is diagnosed by the following characteristics:(i) two distantly separated midventral rows that are morphogenetically formed with an urostylid mode;(ii) absence of the "frontoterminal row",which is formed from the posterior-most frontoventral-transverse cirral anlage in all other typical urostylids;(iii) numerous caudal cirri that derive from each of the dorsal kinety anlagen;(iv) right marginal row that has a unique de novo origin;and (v) inhabiting periphytic communities.The validity of the suborder Pseudoamphisiellina is firmly supported by molecular data.