Epithelial-to-mesenchymal transition (EMT) is a critical event in the progression toward cancer metastasis. The intermediate filament protein vimentin is an important marker of EMT and a requisite ...regulator of mesenchymal cell migration. However, it is not known how vimentin functionally contributes to cancer cell invasion. Here, we report that ectopic expression of oncogenic H-Ras-V12G and Slug induces vimentin expression and migration in pre-malignant breast epithelial cells. Conversely, vimentin expression is necessary for Slug- or H-Ras-V12G-induced EMT-associated migration. Furthermore, silencing of vimentin in breast epithelial cells results in specific changes in invasiveness-related gene expression including upregulation of RAB25 (small GTPase Rab25) and downregulation of AXL (receptor tyrosine kinase Axl), PLAU (plasminogen activator, urokinase) and ITGB4 (integrin β4-subunit). Importantly, gene expression profiling analyses reveal that vimentin expression correlates positively/negatively with these genes also in multiple breast cancer cell lines and breast cancer patient samples. Focusing on the tyrosine kinase Axl, we show that induction of vimentin by EMT is associated with upregulation of Axl expression and that Axl enhances the migratory activity of pre-malignant breast epithelial cells. Using null and knock-down cells and overexpression models, we also show that regulation of breast cancer cell migration in two- and three-dimensional matrices by vimentin is Axl- dependent and that Axl functionally contributes to lung extravasation of breast cancer cells in mice. In conclusion, our data show that vimentin functionally contributes to EMT and is required for induction of Axl expression. Moreover, these results provide a molecular explanation for vimentin-dependent cancer cell migration during EMT by identifying Axl as a key proximal component in this process.
Marine spatial planning relies on detailed spatial information of marine areas to ensure effective conservation of species. To enhance our understanding of marine habitat use by the highly pelagic ...Bermuda petrel
Pterodroma cahow
, we deployed GPS tags on 6 chick-rearing adults in April 2019 and constructed a habitat suitability model using locations classified as foraging to explore functional responses to a selection of marine environmental variables. We defined 15 trips for 5 individuals, ranging from 1-6 trips per bird, that included both short and long foraging excursions indicative of a dual foraging strategy that optimizes chick feeding and self maintenance. The maximum distance birds flew from Bermuda during foraging trips ranged from 61 to 2513 km (total trip lengths: 186-14051 km). Behaviourally deduced foraging habitat was best predicted at shorter distances from the colony, under warmer sea surface temperature, greater sea surface height, and in deeper water compared to transiting locations; our model results indicated that suitable foraging habitat exists beyond the core home range of the population, as far north as the highly productive Gulf Stream frontal system, and within the territorial waters of both the USA and Canada. Our results are crucial to inform management decisions and international conservation efforts by better identifying potential threats encountered at sea by this globally rare seabird and highlighting jurisdictions potentially responsible for mitigating those threats.
Seabirds and other marine animals are at risk from anthropogenic activities that target them directly and those that can harm them incidentally. We integrate year‐round tracking and vessel studies to ...assess risks for a globally important seabird population in the North‐West Atlantic. The eastern Canadian Grand Bank has a rich and diverse food web that supports an abundance of apex predators. Major resource extraction industries (hydrocarbon production and fisheries) operate in the area, and, in addition to shipping and hunting, pose risks for marine birds. Understanding the relative risks has been hampered by poor information on bird distribution at sea. Here, we deployed global location sensors (loggers or geolocators) on common murres Uria aalge at Funk Island, the species' largest North American breeding colony. Adults (n=10) were resident on the Grand Bank and in adjacent pelagic waters year round. Within 10 days of leaving the colony, males dispersed offshore (<50°W), south–south‐east of Funk Island. Females departed later and spent 10–47 days in coastal waters before moving offshore. All birds were in the vicinity of offshore oil platforms during November and December, but remained outside the area of the coastal Newfoundland and Labrador murre hunt. Three of six tracked females, but only one of four tracked males moved closer to shore during January and February where vulnerability to the hunt may have increased. Vessel‐based surveys confirmed the importance of offshore, shelf‐edge habitats for murres in winter. Our results highlight the relative risk to wintering murres from different human activities, providing a sound scientific rationale for focusing conservation and management actions. This information is particularly timely given the continued expansion of deep‐water drilling in the North‐West Atlantic and increasing risk of oil pollution for seabirds attracted to platforms.
I used a supplemental feeding experiment to determine whether adult Tufted Puffins (Fratercula cirrhata) would decrease provisioning effort in response to a reduction in nestling nutritional ...requirements, and to investigate the relationship between parental provisioning effort and timing of fledging. As predicted, parents of the supplemented nestlings decreased the number of provisioning trips but did not alter bill-load size or prey composition. Supplemental feeding significantly increased the growth of the culmen and tarsus but had no detectable effect on wing growth or body mass of chicks. Supplemented nestlings fledged significantly older than control nestlings. The maximum mass attained by the nestling and the age at which mass peaked also influenced timing of fledging. These results demonstrate that fledging age is influenced by both the energy provided by the parents and the nestling's developmental state and are consistent with the hypothesis that nestlings time their departure from the nest based on the costs and benefits of remaining.
There is a great demand for the improvement of mammalian cell production systems such that they can compete economically with their prokaryotic counterparts. Of a number of parameters that need to be ...explored to accomplish this we have tested the effects of different signal peptides on the synthesis and secretion of
Gaussia princeps luciferase in mammalian cells. A series of plasmids were transfected into CHO cells where the coding region for the marine luciferase was fused to the signal peptide coding regions derived from different sources. Both cell extracts and medium samples were analysed for luciferase activity. When the native
Gaussia luciferase signal sequence in the vector was substituted by that from human interleukin-2 or albumin then the amount of active recombinant protein produced was substantially reduced, both in transiently and stably transfected cells. Western blotting showed that enzyme activity and protein levels mirrored one another. The major decrease in luciferase activity was shown not to be a result of decreased mRNA levels, indicating the involvement of a post-transcriptional event. When the coding region of human endostatin was fused to that of the
Gaussia luciferase signal peptide then an elevated level of secreted endostatin was observed compared to when that of the albumin signal peptide was used. Stable transfection of HepG2 cells with the different signal peptide constructs gave essentially the same results as seen in CHO cells. The overall results indicate that the choice of signal peptide can be imperative to ensure an optimal synthesis and secretion of a recombinant protein in a mammalian cell culture system.
We examined nest-site selection and nesting success in Saltmarsh Sharp-tailed (Ammodramus caudacutus) and Seaside Sparrows (A. maritimus), at seven sites in Connecticut. We found 160 Saltmarsh ...Sharp-tailed Sparrow nests and 23 Seaside Sparrow nests, and compared characteristics of their locations to each other and to random locations. We tracked success of all nests, quantified nest productivity and causes of nest losses, and tested for habitat differences between successful and unsuccessful nests. Saltmarsh Sharp-tailed Sparrows nested in higher than average locations, where the vegetation was taller and more dense than at random locations, where there was a deep layer of thatch, and where saltmeadow cordgrass (Spartina patens) dominated the vegetation. There was little evidence that habitat characteristics influenced the success of nesting birds, but the timing of nest initiation relative to spring tides was important. Seaside Sparrow nests occurred in even taller vegetation, that was more sparse than average and dominated by the tall form of smooth cordgrass (S. alterniflora). Habitat influenced the success of Seaside Sparrow nests, but timing did not; on average, successful nests occurred in taller vegetation. Model comparisons suggest that vegetation structure influences site selection more than species composition or inherent differences among marshes. Overall, our results indicate that nest flooding is a major threat to successful reproduction in both species, but they have different strategies to avoid flooding. Saltmarsh Sharp-tailed Sparrows time their reproduction to avoid especially high tides, while Seaside Sparrows avoid flooding spatially by nesting in tall vegetation.
Anomalously warm sea-surface temperatures (SSTs) are associated with interannual and decadal variability as well as with long-term climate changes indicative of global warming. Such oscillations ...could precipitate changes in a variety of oceanic processes to affect marine species worldwide. As global temperatures continue to rise, it will be critically important to be able to predict the effects of such changes on species' abundance, distribution, and ecological relationships so as to identify vulnerable populations. Off the coast of British Columbia, warm SSTs have persisted through the last two decades. Based on 16 years of reproductive data collected between 1975 and 2002, we show that the extreme variation in reproductive performance exhibited by tufted puffins (Fratercula cirrhata) was related to changes in SST both within and among seasons. Especially warm SSTs corresponded with drastically decreased growth rates and fledging success of puffin nestlings. Puffins may partially compensate for within-season changes associated with SST by adjusting their breeding phenology, yet our data also suggest that they are highly vulnerable to the effects of climate change at this site and may serve as a valuable indicator of biological change in the North Pacific. Further and prolonged increases in ocean temperature could make Triangle Island, which contains the largest tufted puffin colony in Canada, unsuitable as a breeding site for this species.
I used a supplemental feeding experiment to determine whether adult Tufted Puffins (Fratercula cirrhata) would decrease provisioning effort in response to a reduction in nestling nutritional ...requirements, and to investigate the relationship between parental provisioning effort and timing of fledging. As predicted, parents of the supplemented nestlings decreased the number of provisioning trips but did not alter bill-load size or prey composition. Supplemental feeding significantly increased the growth of the culmen and tarsus but had no detectable effect on wing growth or body mass of chicks. Supplemented nestlings fledged significantly older than control nestlings. The maximum mass attained by the nestling and the age at which mass peaked also influenced timing of fledging. These results demonstrate that fledging age is influenced by both the energy provided by the parents and the nestling's developmental state and are consistent with the hypothesis that nestlings time their departure from the nest based on the costs and benefits of remaining.
We examined nest-site selection and nesting success in Saltmarsh Sharp-tailed (Ammodramus caudacutus) and Seaside Sparrows (A. maritimus), at seven sites in Connecticut. We found 160 Saltmarsh ...Sharp-tailed Sparrow nests and 23 Seaside Sparrow nests, and compared characteristics of their locations to each other and to random locations. We tracked success of all nests, quantified nest productivity and causes of nest losses, and tested for habitat differences between successful and unsuccessful nests. Saltmarsh Sharp-tailed Sparrows nested in higher than average locations, where the vegetation was taller and more dense than at random locations, where there was a deep layer of thatch, and where saltmeadow cordgrass (Spartina patens) dominated the vegetation. There was little evidence that habitat characteristics influenced the success of nesting birds, but the timing of nest initiation relative to spring tides was important. Seaside Sparrow nests occurred in even taller vegetation, that was more sparse than average and dominated by the tall form of smooth cordgrass (S. alterniflora). Habitat influenced the success of Seaside Sparrow nests, but timing did not; on average, successful nests occurred in taller vegetation. Model comparisons suggest that vegetation structure influences site selection more than species composition or inherent differences among marshes. Overall, our results indicate that nest flooding is a major threat to successful reproduction in both species, but they have different strategies to avoid flooding. Saltmarsh Sharp-tailed Sparrows time their reproduction to avoid especially high tides, while Seaside Sparrows avoid flooding spatially by nesting in tall vegetation.
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