Centrosome-independent mitotic spindle formation in vertebrates

AbstractBackground: In cells lacking centrosomes, the microtubule-organizing activity of the centrosome is substituted for by the combined action of chromatin and molecular motors. The question of whether a centrosome-independent pathway for spindle formation exists in vertebrate somatic cells, which always contain centrosomes, remains unanswered, however. By a combination of labeling with green fluorescent protein (GFP) and laser microsurgery we have been able to selectively destroy centrosomes in living mammalian cells as they enter mitosis.Results: We have established a mammalian cell line in which the boundaries of the centrosome are defined by the constitutive expression of γ-tubulin–GFP. This feature allows us to use laser microsurgery to selectively destroy the centrosomes in living cells. Here we show that this method can be used to reproducibly ablate the centrosome as a functional entity, and that after destruction the microtubules associated with the ablated centrosome disassemble. Depolymerization–repolymerization experiments reveal that microtubules form in acentrosomal cells randomly within the cytoplasm. When both centrosomes are destroyed during prophase these cells form a functional bipolar spindle. Surprisingly, when just one centrosome is destroyed, bipolar spindles are also formed that contain one centrosomal and one acentrosomal pole. Both the polar regions in these spindles are well focused and contain the nuclear structural protein NuMA. The acentrosomal pole lacks pericentrin, γ-tubulin, and centrioles, however.Conclusions: These results reveal, for the first time, that somatic cells can use a centrosome-independent pathway for spindle formation that is normally masked by the presence of the centrosome. Furthermore, this mechanism is strong enough to drive bipolar spindle assembly even in the presence of a single functional centrosome

Assessment of the SEEV Model to Predict Attention Allocation at Intersections During Simulated Driving

We attempted to model attention allocation of experienced drivers using the SEEV model. Unlike previous attempts, the present work looked at attention to entities (vehicles, signs, traffic control devices) in the outside world rather than considering the outside world as a unitary construct. Model parameters were generated from rankings of entities by experienced drivers. Experienced drivers drove a scenario that included a number of intersections interspersed with stretches of straight road. The intersections included non-hazard events. Eye movements were monitored during the driving session. The results of fitting the observed eye movement data to our SEEV model were poor, and were no better than fitting the data to a randomized SEEV model. A number of explanations for this are discussed

Hominin tracks in southern Africa: a review and an approach to identification

Three Late Pleistocene hominin tracksites have been reported from coastal aelioanites in South Africa. Two have been dated to 124 ka and 117 ka , and the third is inferred to be 90 ka. There are no other globally reported sites for probable Homo sapiens tracks older than 46 ka. Given this documented record, a search for further hominin tracksites in southern Africa may well yield additional positive results. However, this is a field that demands scientific rigour, as false positive tracksites (pseudotracks) may occur. Criteria have been developed for the identification of fossil vertebrate tracks and hominin tracks, but these are specific neither to southern Africa nor to aeolianites.An important caveat is that the tracks of shod humans would not fulfil these criteria. Preservation of tracks varies with facies and is known to be suboptimal in aeolianites. An analysis of the tracks from the three documented South African sites, along with pseudotracks and tracks of questionable provenance, allows for the proposal and development of guidelines for fossil hominin track identification that are of specific relevance to southern Africa. Such guidelines have broader implications for understanding the constraints that track preservation and substrate have on identifying diagnostic morphological features.Palaeontological Scientific Trust (PAST)JNC201

Nasal Congestion on the International Space Station

No abstract availabl

Development of the morpholino gene knockdown technique in Fundulus heteroclitus : a tool for studying molecular mechanisms in an established environmental model

Author Posting. © Elsevier B.V., 2008. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Aquatic Toxicology 87 (2008): 289-295, doi:10.1016/j.aquatox.2008.02.010.A significant challenge in environmental toxicology is that many genetic and genomic tools available in laboratory models are not developed for commonly used environmental models. The Atlantic killifish (Fundulus heteroclitus) is one of the most studied teleost environmental models, yet few genetic or genomic tools have been developed for use in this species. The advancement of genetic and evolutionary toxicology will require that many of the tools developed in laboratory models be transferred into species more applicable to environmental toxicology. Antisense morpholino oligonucleotide (MO) gene knockdown technology has been widely utilized to study development in zebrafish and has been proven to be a powerful tool in toxicological investigations through direct manipulation of molecular pathways. To expand the utility of killifish as an environmental model, MO gene knockdown technology was adapted for use in Fundulus. Morpholino microinjection methods were altered to overcome the significant differences between these two species. Morpholino efficacy and functional duration were evaluated with molecular and phenotypic methods. A cytochrome P450-1A (CYP1A) MO was used to confirm effectiveness of the methodology. For CYP1A MO-injected embryos, a 70% reduction in CYP1A activity, a 86% reduction in total CYP1A protein, a significant increase in β-naphthoflavone-induced teratogenicity, and estimates of functional duration (50% reduction in activity 10 dpf, and 86% reduction in total protein 12 dpf) conclusively demonstrated that MO technologies can be used effectively in killifish and will likely be just as informative as they have been in zebrafish.This work was funded in part by the National Institute of Environmental Health Sciences through the Duke Superfund Basic Research Center (P42ES010356), the Boston University Superfund Basic Research Program (P42ES007381), and the Duke Integrated Toxicology and Environmental Health Program (ES-T32-0007031). Additional support was provided by a U.S. Environmental Protection Agency STAR fellowship awarded to C.R.F