Mitotic Spindle Proteomics in Chinese Hamster Ovary Cells

Mitosis is a fundamental process in the development of all organisms. The mitotic spindle guides the cell through mitosis as it mediates the segregation of chromosomes, the orientation of the cleavage furrow, and the progression of cell division. Birth defects and tissue-specific cancers often result from abnormalities in mitotic events. Here, we report a proteomic study of the mitotic spindle from Chinese Hamster Ovary (CHO) cells. Four different isolations of metaphase spindles were subjected to Multi-dimensional Protein Identification Technology (MudPIT) analysis and tandem mass spectrometry. We identified 1155 proteins and used Gene Ontology (GO) analysis to categorize proteins into cellular component groups. We then compared our data to the previously published CHO midbody proteome and identified proteins that are unique to the CHO spindle. Our data represent the first mitotic spindle proteome in CHO cells, which augments the list of mitotic spindle components from mammalian cells

Variation in Symbiodinium ITS2 Sequence Assemblages among Coral Colonies

Endosymbiotic dinoflagellates in the genus Symbiodinium are fundamentally important to the biology of scleractinian corals, as well as to a variety of other marine organisms. The genus Symbiodinium is genetically and functionally diverse and the taxonomic nature of the union between Symbiodinium and corals is implicated as a key trait determining the environmental tolerance of the symbiosis. Surprisingly, the question of how Symbiodinium diversity partitions within a species across spatial scales of meters to kilometers has received little attention, but is important to understanding the intrinsic biological scope of a given coral population and adaptations to the local environment. Here we address this gap by describing the Symbiodinium ITS2 sequence assemblages recovered from colonies of the reef building coral Montipora capitata sampled across Kāne'ohe Bay, Hawai'i. A total of 52 corals were sampled in a nested design of Coral Colony(Site(Region)) reflecting spatial scales of meters to kilometers. A diversity of Symbiodinium ITS2 sequences was recovered with the majority of variance partitioning at the level of the Coral Colony. To confirm this result, the Symbiodinium ITS2 sequence diversity in six M. capitata colonies were analyzed in much greater depth with 35 to 55 clones per colony. The ITS2 sequences and quantitative composition recovered from these colonies varied significantly, indicating that each coral hosted a different assemblage of Symbiodinium. The diversity of Symbiodinium ITS2 sequence assemblages retrieved from individual colonies of M. capitata here highlights the problems inherent in interpreting multi-copy and intra-genomically variable molecular markers, and serves as a context for discussing the utility and biological relevance of assigning species names based on Symbiodinium ITS2 genotyping

The recovery of coral genetic diversity in the Sunda Strait following the 1883 eruption of Krakatau

Surveys of microsatellite variation show that genetic diversity has largely recovered in two reef-building corals, Pocillopora damicornis and Seriatopora hystrix (Scleractinia: Pocilloporidae), on reefs which were decimated by the eruption of the volcano Krakatau in 1883. Assignment methods and gene flow estimates indicate that the recolonization of Krakatau occurred mainly from the closest upstream reef system, Pulau Seribu, but that larval input from other regions has also occurred. This pattern is clearer in S. hystrix, which is traditionally the more dispersal- limited species. Despite these observed patterns of larval dispersal, self-recruitment appears to now be the most important factor in supplying larvae to coral populations in Krakatau. This suggests that the colonization of devastated reefs can occur quickly through larval dispersal; however, their survival requires local sources of larvae for self-recruitment. This research supports the observation that the recovery of genetic diversity in coral reef animals can occur on the order of decades and centuries rather than millennia. Conservation measures aimed at sustaining coral reef populations in Krakatau and elsewhere should include both the protection of upstream source populations for larval replenishment should disaster occur as well as the protection of large adult colonies to serve as local larval source

Coral reefs Corals' adaptive response to climate change

The long-term response of coral reefs to climate change depends on the ability of reef-building coral symbioses to adapt or acclimatize to warmer temperatures, but there has been no direct evidence that such a response can occur. Here we show that corals containing unusual algal symbionts that are thermally tolerant and commonly associated with high-temperature environments are much more abundant on reefs that have been severely affected by recent climate change. This adaptive shift in symbiont communities indicates that these devastated reefs could be more resistant to future thermal stress, resulting in significantly longer extinction times for surviving corals than had been previously assumed

Ten polymorphic STR loci in the cosmopolitan reef coral, Pocillopora damicornis

We report the development of 10 polymorphic molecular markers containing short tandem repeats in the cosmopolitan reef‐building coral, Pocillopora damicornis, an important model species for coral health, physiology, ecology, and genetics. The availability of polymorphic DNA markers in P. damicornis can act as impetus for investigations into inheritance and population genetics, as well as novel investigations into host‐symbiont ecology and evolution. Coral bleaching and gene flow studies performed with these markers can have direct conservation implications

Strong genetic structure among coral populations within a conservation priority region, the Birds Head Seascape (Papua and West Papua, Indonesia)

Marine Protected Areas (MPAs) are widely considered to be one of the best strategies available for protecting species diversity and ecosystem processes in marine environments, particularly in developing, tropical nations. While data on connectivity and genetic structure of marine populations are critical to designing appropriately sized and spaced networks of MPAs, such data are rarely available. Here we present an assessment of genetic structure in reef-building corals from Papua and West Papua, Indonesia, among the most biologically diverse and least disturbed coral reef regions in the world, and the focus of the multi-institutional Birds Head Seascape initiative to design and implement a functional network of MPAs. Microsatellite variation was assessed within and among populations of Pocillopora damicornis (Linnaeus , 1758) and Seriatopora hystrix (Dana 1846) (family: Pocilloporidae) from three regions, each currently under a different conservation regime: Teluk Cenderawasih, Raja Ampat, and southwest Papua. Analyses of molecular variance, assignment tests, and genetical bandwidth mapping revealed significant local-scale structure in both species, and a lack of regional filters to gene flow. Overall, P. damicornis populations were less structured ( F ST = 0.139, p < 0.00001) than those of S. hystrix ( F ST = 0.357, p < 0.00001). In order to maintain connectivity within and among regions, coral reef conservation on the local scale is needed. These data have been directly applied to the design of a MPA network in the Bird’s Head Seascape

Morphogenesis of Human Placental Chorionic Villi:Cytoskeletal, Syncytioskeletal and Extracellular Matrix Proteins

Immunocytochemical and ultrastructural methods were used to investigate the distribution of a family of structural proteins in the human placenta near term. These reveal the distribution of cytoskeletal and `syncytioskeletal' components that may account for some of the more obvious micromorphological features of placental structure. In the syncytiotrophoblast a potentially supporting structure `the syncytioskeletal layer' is described. It is an apparently continuous and complex polymeric network covering the villous tree, a surface of the order of 10 m$^{2}$ in area in the full term placenta (Aherne & Dunnill 1966). It is suggested that this layer plays a part in morphogenesis of the villous tree

Characteristics of cells derived from the girdle region of the pre-implantation blastocyst of the donkey

The establishment of a monolayer culture of cells derived from the girdle region of a 34-day-old donkey conceptus is described. These cells have had over 100 repeated passages in culture. Low levels of pregnant mares' serum gonadotrophin (PMSG, eCG) could be detected in the cells by indirect immunofluorescence using some monoclonal anti-eCG antibodies, but the cells did not secrete eCG as measured by radioimmunoassay or inhibition of haemagglutination. There was marked nuclear polymorphism with binucleate and occasional multinucleate cells. The cells were strongly reactive with wheatgerm agglutinin and concanavalin A suggesting the synthesis of many glycosylated products. Some cells were reactive with antisera to prekeratin, others with antisera to vimentin. The cells also contained actin (showing peculiar intercellular communications), -actinin and tubulin. They were able to metabolize certain steroid precursors, but there was no definitive evidence for the presence of aromatase or 5-3-hydroxysteroid dehydrogenase in these cells. This cell line appears to resemble trophectodermal girdle epithelium at a stage of development prior to the onset of eCG production, and may be useful in studies on the control of expression of this substance