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1.
Mol Phylogenet Evol ; 160: 107127, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33667632

RESUMO

The assembly of the Australian arid zone biota has long fascinated biogeographers. Covering over two-thirds of the continent, Australia's vast arid zone biome is home to a distinctive fauna and flora, including numerous lineages which have diversified since the Eocene. Tracing the origins and speciation history of these arid zone taxa has been an ongoing endeavour since the advent of molecular phylogenetics, and an increasing number of studies on invertebrate animals are beginning to complement a rich history of research on vertebrate and plant taxa. In this study, we apply continent-wide genetic sampling and one of the largest phylogenetic data matrices yet assembled for a genus of Australian spiders, to reconstruct the phylogeny and biogeographic history of the open-holed trapdoor spider genus Aname L. Koch, 1873. This highly diverse lineage of Australian mygalomorph spiders has a distribution covering the majority of Australia west of the Great Dividing Range, but apparently excluding the high rainfall zones of eastern Australia and Tasmania. Original and legacy sequences were obtained for three mtDNA and four nuDNA markers from 174 taxa in seven genera, including 150 Aname specimen terminals belonging to 102 species-level operational taxonomic units, sampled from 32 bioregions across Australia. Reconstruction of the phylogeny and biogeographic history of Aname revealed three radiations (Tropical, Temperate-Eastern and Continental), which could be further broken into eight major inclusive clades. Ancestral area reconstruction revealed the Pilbara, Monsoon Tropics and Mid-West to be important ancestral areas for the genus Aname and its closest relatives, with the origin of Aname itself inferred in the Pilbara bioregion. From these origins in the arid north-west of Australia, our study found evidence for a series of subsequent biome transitions in separate lineages, with at least eight tertiary incursions back into the arid zone from more mesic tropical, temperate or eastern biomes, and only two major clades which experienced widespread (primary) in situ diversification within the arid zone. Based on our phylogenetic results, and results from independent legacy divergence dating studies, we further reveal the importance of climate-driven biotic change in the Miocene and Pliocene in shaping the distribution and composition of the Australian arid zone biota, and the value of continent-wide studies in revealing potentially complex patterns of arid zone diversification in dispersal-limited invertebrate taxa.


Assuntos
Clima Desértico , Filogenia , Filogeografia , Aranhas/classificação , Aranhas/genética , Animais , Austrália , DNA Mitocondrial/genética
2.
Ecotoxicology ; 30(4): 560-574, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33770305

RESUMO

Rats and mice can damage food and agricultural products as well as transmit diseases, thereby requiring control of their numbers. Application of Second Generation Anticoagulant Rodenticides (SGARs) often reduces rodent numbers locally. However, predators eating rodents, including non-target species, that have consumed SGARs may be secondarily exposed and potentially lethally poisoned. Here we study whether SGARs may have contributed to the widespread population declines of a rodent-eating raptor, the Common Kestrel (Falco tinnunculus) in the UK. We show that 161 (66.8%) of the 241 Kestrels submitted for ecotoxicology tests between 1997 and 2012 had detectable levels of at least one SGAR in their livers. Adult Kestrels had significantly higher prevalence of SGARs than juveniles, suggesting accumulation of SGARs through time. The prevalence and concentrations of individual SGARs in Kestrels were significantly higher in England than in Scotland. SGAR prevalence in Kestrels were positively associated with some land cover types, primarily arable cereals and broad-leaved woodland, and negatively associated with mainly mean elevation, probably reflecting variation in SGAR usage across land cover types. By using volunteer-collected data on national Kestrel abundance 1997-2012, we show that there is a negative correlation between the Kestrel population index in a specific year and the concentration of bromadialone as well as the total SGAR concentration in the same year. Although correlative, this is the first study to provide evidence for a potential population-limiting effect of SGARs on a raptor.


Assuntos
Falconiformes , Aves Predatórias , Rodenticidas , Animais , Anticoagulantes , Inglaterra , Camundongos , Ratos , Rodenticidas/toxicidade
3.
Glob Chang Biol ; 26(7): 3771-3787, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32350939

RESUMO

Climate change and anthropogenic nitrogen deposition are widely regarded as important drivers of environmental change in alpine habitats. However, due to the difficulties working in high-elevation mountain systems, the impacts of these drivers on alpine breeding species have rarely been investigated. The Eurasian dotterel (Charadrius morinellus) is a migratory wader, which has been the subject of uniquely long-term and spatially widespread monitoring effort in Scotland, where it breeds in alpine areas in dwindling numbers. Here we analyse data sets spanning three decades, to investigate whether key potential drivers of environmental change in Scottish mountains (snow lie, elevated summer temperatures and nitrogen deposition) have contributed to the population decline of dotterel. We also consider the role of rainfall on the species' wintering grounds in North Africa. We found that dotterel declines-in both density and site occupancy of breeding males-primarily occurred on low and intermediate elevation sites. High-elevation sites mostly continued to be occupied, but males occurred at lower densities in years following snow-rich winters, suggesting that high-elevation snow cover displaced dotterel to lower sites. Wintering ground rainfall was positively associated with densities of breeding males two springs later. Dotterel densities were reduced at low and intermediate sites where nitrogen deposition was greatest, but not at high-elevation sites. While climatic factors explained variation in breeding density between years, they did not seem to explain the species' uphill retreat and decline. We cannot rule out the possibility that dotterel have increasingly settled on higher sites previously unavailable due to extensive snow cover, while changes associated with nitrogen deposition may also have rendered lower lying sites less suitable for breeding. Causes of population and range changes in mountain-breeding species are thus liable to be complex, involving multiple anthropogenic drivers of environmental change acting widely across annual and migratory life cycles.


Assuntos
Aves , Mudança Climática , África do Norte , Animais , Cruzamento , Masculino , Dinâmica Populacional , Escócia , Estações do Ano
4.
Cladistics ; 36(6): 543-568, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34618949

RESUMO

In the trapdoor spider genus Euoplos Rainbow & Pulleine (tribe Euoplini), it was discovered recently that two divergent lineages occur in sympatry in eastern Australia. This challenged the monogeneric classification of the tribe and, in combination with inadequate taxonomic descriptions of some species, precluded comprehensive taxonomic revision. To resolve these issues, we conducted a total-evidence cladistic analysis on a largely undescribed continental fauna-the first such analysis on a group of Australian Mygalomorphae. We combined multilocus molecular data and/or morphological and behavioural data from all known species from eastern Australia (described and undescribed), plus a subset of Western Australian species, to produce a phylogeny for the tribe. We mapped morphological/behavioural characters onto this to identify clade-specific diagnostic characters, and applied these data to a generic reclassification of the tribe. We recovered two sympatric lineages in the Euoplini (the "wafer-door" and "plug-door/palisade" lineages), and revealed the phylogenetic position of all known eastern Australian species within these. Character mapping revealed morphological and behavioural (burrow architecture) features that allow diagnosis of the lineages and clades within them. We erect a new genus, Cryptoforis gen.n., to represent the wafer-door lineage, describe the type species, Cryptoforis hughesae sp.n., and transfer two species from Euoplos to Cryptoforis: C. tasmanica (Hickman, 1928) and C. victoriensis (Main, 1995). This study resolves phylogenetic structure within the Euoplini, and characterizes clades within the tribe to facilitate future taxonomic revisions. It also demonstrates that, whereas male morphology is more informative, female morphological characters relating to genitalia and the scopulation/spination of the anterior legs display phylogenetic signal in the Euoplini, highlighting the subtle nature of informative female characters in mygalomorph spiders.

5.
Mol Phylogenet Evol ; 122: 157-165, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29428510

RESUMO

Spiders of the infraorder Mygalomorphae are fast becoming model organisms for the study of biogeography and speciation. However, these spiders can be difficult to study in the absence of fundamental life history information. In particular, their cryptic nature hinders comprehensive sampling, and linking males with conspecific females can be challenging. Recently discovered differences in burrow entrance architecture and male morphology indicated that these challenges may have impeded our understanding of the trapdoor spider genus Euoplos in Australia's eastern mesic zone. We investigated the evolutionary significance of these discoveries using a multi-locus phylogenetic approach. Our results revealed the existence of a second, previously undocumented, lineage of Euoplos in the eastern mesic zone. This new lineage occurs in sympatry with a lineage previously known from the region, and the two are consistently divergent in their burrow entrance architecture and male morphology, revealing the suitability of these characters for use in phylogenetic studies. Divergent burrow entrance architecture and observed differences in microhabitat preferences are suggested to facilitate sympatry and syntopy between the lineages. Finally, by investigating male morphology and plotting it onto the phylogeny, we revealed that the majority of Euoplos species remain undescribed, and that males of an unnamed species from the newly discovered lineage had historically been linked, erroneously, to a described species from the opposite lineage. This paper clarifies the evolutionary relationships underlying life history diversity in the Euoplos of eastern Australia, and provides a foundation for urgently needed taxonomic revision of this genus.


Assuntos
Filogenia , Aranhas/anatomia & histologia , Aranhas/classificação , Animais , Austrália , Evolução Biológica , Complexo IV da Cadeia de Transporte de Elétrons/genética , Feminino , Masculino , Análise de Sequência de DNA , Aranhas/genética , Simpatria
6.
J Hered ; 107(5): 413-22, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27225934

RESUMO

In 1993, a population of freshwater glass shrimp (Paratya australiensis) was translocated from Kilcoy Creek to Branch Creek in the Conondale Range, Queensland. Subsequent genetic analysis revealed that the translocated and resident shrimp belonged to different mitochondrial DNA (mtDNA) lineages that were capable of hybridizing. Monitoring of the pools along Branch Creek up until 2002 suggested that the translocated lineage had an advantage in upstream pools and the resident lineage dominated downstream. Differential temperature tolerance and hybridization barriers such as hybrid inviability and mate selection were factors proposed to explain hybrid zone structure. The major objective of this study was to combine nuclear and mtDNA markers to identify the structure of the hybrid zone in 2013 and identify any changes that had occurred since 2002. Specifically, we used genetic data to test for evidence of hybrid zone movement and used the inbreeding coefficient (F IS) to investigate whether mating was random in the contact zone where hybridization barriers could be present. The results revealed that the hybrid zone center has shifted 510 m downstream since 2002. Increased rainfall in the region since 2010 could have facilitated this. Secondly, mating appears significantly nonrandom in the pools where both lineages occur, supporting the existence of partial hybridization barriers. This study reveals a complex and dynamic hybrid zone and exemplifies why multiple temporal studies are necessary to understand hybrid zone structure.


Assuntos
Crustáceos/genética , Genética Populacional , Hibridização Genética , Animais , Austrália , DNA Mitocondrial , Evolução Molecular , Frequência do Gene , Variação Genética , Genótipo , Repetições de Microssatélites , Isolamento Reprodutivo
7.
Trends Ecol Evol ; 39(1): 89-100, 2024 01.
Artigo em Inglês | MEDLINE | ID: mdl-38114339

RESUMO

We present the results of our 15th horizon scan of novel issues that could influence biological conservation in the future. From an initial list of 96 issues, our international panel of scientists and practitioners identified 15 that we consider important for societies worldwide to track and potentially respond to. Issues are novel within conservation or represent a substantial positive or negative step-change with global or regional extents. For example, new sources of hydrogen fuel and changes in deep-sea currents may have profound impacts on marine and terrestrial ecosystems. Technological advances that may be positive include benchtop DNA printers and the industrialisation of approaches that can create high-protein food from air, potentially reducing the pressure on land for food production.


Assuntos
Biodiversidade , Ecossistema , Conservação dos Recursos Naturais , Previsões , Alimentos
8.
Ecol Evol ; 13(1): e9706, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36636427

RESUMO

Understanding the drivers of morphological convergence requires investigation into its relationship with behavior and niche space, and such investigations in turn provide insights into evolutionary dynamics, functional morphology, and life history. Mygalomorph spiders (trapdoor spiders and their kin) have long been associated with high levels of morphological homoplasy, and many convergent features can be intuitively associated with different behavioral niches. Using genus-level phylogenies based on recent genomic studies and a newly assembled matrix of discrete behavioral and somatic morphological characters, we reconstruct the evolution of burrowing behavior in the Mygalomorphae, compare the influence of behavior and evolutionary history on somatic morphology, and test hypotheses of correlated evolution between specific morphological features and behavior. Our results reveal the simplicity of the mygalomorph adaptive landscape, with opportunistic, web-building taxa at one end, and burrowing/nesting taxa with structurally modified burrow entrances (e.g., a trapdoor) at the other. Shifts in behavioral niche, in both directions, are common across the evolutionary history of the Mygalomorphae, and several major clades include taxa inhabiting both behavioral extremes. Somatic morphology is heavily influenced by behavior, with taxa inhabiting the same behavioral niche often more similar morphologically than more closely related but behaviorally divergent taxa, and we were able to identify a suite of 11 somatic features that show significant correlation with particular behaviors. We discuss these findings in light of the function of particular morphological features, niche dynamics within the Mygalomorphae, and constraints on the mygalomorph adaptive landscape relative to other spiders.

9.
Trends Ecol Evol ; 38(1): 96-107, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36460563

RESUMO

We present the results of our 14th horizon scan of issues we expect to influence biological conservation in the future. From an initial set of 102 topics, our global panel of 30 scientists and practitioners identified 15 issues we consider most urgent for societies worldwide to address. Issues are novel within biological conservation or represent a substantial positive or negative step change at global or regional scales. Issues such as submerged artificial light fisheries and accelerating upper ocean currents could have profound negative impacts on marine or coastal ecosystems. We also identified potentially positive technological advances, including energy production and storage, improved fertilisation methods, and expansion of biodegradable materials. If effectively managed, these technologies could realise future benefits for biological diversity.


Assuntos
Conservação dos Recursos Naturais , Ecossistema , Biodiversidade , Previsões , Pesqueiros
10.
Trends Ecol Evol ; 37(1): 95-104, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34809998

RESUMO

We present the results of our 13th annual horizon scan of issues likely to impact on biodiversity conservation. Issues are either novel within the biological conservation sector or could cause a substantial step-change in impact, either globally or regionally. Our global panel of 26 scientists and practitioners identified 15 issues that we believe to represent the highest priorities for tracking and action. Many of the issues we identified, including the impact of satellite megaconstellations and the use of long-distance wireless energy transfer, have both elements of threats and emerging opportunities. A recent state-sponsored application to commence deep-sea mining represents a significant step-change in impact. We hope that this horizon scan will increase research and policy attention on the highlighted issues.


Assuntos
Biodiversidade , Conservação dos Recursos Naturais , Saúde Global/tendências , Animais , Políticas
11.
Zootaxa ; 4861(1): zootaxa.4861.1.5, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-33055870

RESUMO

The D'Aguilar Range of subtropical south-eastern Queensland (Australia), harbours an upland rainforest biota characterised by high levels of endemic diversity. Following recent phylogenetic and biogeographic research into the open-holed trapdoor spiders of the genus Namea Raven, 1984 (family Anamidae), remarkable levels of sympatry for a single genus of mygalomorph spiders were recorded from the D'Aguilar Range. It is now known that eight different species in the genus can be found in the D'Aguilar uplands, with five apparently endemic to rainforest habitats. In this paper we present a phylogenetic and taxonomic synopsis of the remarkable anamid fauna of the D'Aguilar Range: a key to the eight species is provided, and four new species of Namea are described (N. gloriosa sp. nov., N. gowardae sp. nov., N. nebo sp. nov. and N. nigritarsus sp. nov.). In shining a spotlight on the mygalomorph spiders of this region, we highlight the D'Aguilar Range as a hotspot of subtropical rainforest diversity, and an area of considerable conservation value.


Assuntos
Floresta Úmida , Aranhas , Animais , Austrália , Filogenia
12.
PLoS One ; 15(2): e0228742, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32050003

RESUMO

We propose a new approach to quantifying a minimum threshold value for the size of an animal population, below which that population might be categorised as having unfavourable status. Under European Union law, the concept of Favourable Conservation Status requires assessment of populations as having favourable or unfavourable status, but quantitative methods for such assessments have not yet been developed. One population threshold that is well established in conservation biology is the minimum viable population (MVP) defined as the size of a small but stable population with an acceptably low risk of extinction within a specified period. Our approach combines this small-population paradigm MVP concept with a multiplier, which is a factor by which the MVP is multiplied to allow for the risk of a sustained future decline. We demonstrate this approach using data on UK breeding bird population sizes. We used 43-year time-series data for 189 species and a qualitative assessment of population trends over almost 200 years for 229 species to examine the prevalence, duration and magnitude of sustained population declines. Our study addressed the problem of underestimation of the duration and magnitude of declines caused by short runs of monitoring data by allowing for the truncation of time series. The multiplier was derived from probability distributions of decline magnitudes within a given period, adjusted for truncation. Over a surveillance period of 100 years, we estimated that there was a 10% risk across species that a sustained population decline of at least sixteen-fold would begin. We therefore suggest that, in this case, a factor of 16 could be used as the multiplier of small-population MVPs to obtain minimum threshold population sizes for favourable status. We propose this 'MVP Multiplier' method as a new and robust approach to obtaining minimum threshold population sizes which integrates the concepts of small-population and declining-population paradigms. The minimum threshold value we propose is intended for use alongside a range of other measures to enable overall assessments of favourable conservation status.


Assuntos
Estatística como Assunto/métodos , Animais , Conservação dos Recursos Naturais , Densidade Demográfica , Prevalência , Fatores de Tempo
13.
Zootaxa ; 4674(3): zootaxa.4674.3.3, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31716002

RESUMO

The open-holed trapdoor spiders of the genus Teyl Main, 1975 from the Pilbara bioregion of Western Australia are investigated. A single endemic species from the southern Pilbara, T. heuretes sp. nov., is newly described, representing the northern-most occurrence of the genus in Australia. Legacy molecular data for Australian Nemesiidae, along with newly generated sequences for all described species of Teyl known from Western Australia, are analysed using Maximum Likelihood methods, providing molecular data for T. heuretes and an expanded phylogenetic assessment of the genus.


Assuntos
Aranhas , Animais , Austrália , Filogenia , Austrália Ocidental
14.
Ambio ; 37(6): 460-5, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18833801

RESUMO

The Scottish Raptor Monitoring Scheme (SRMS) comprises 7 partner organizations and was established in 2002 after i) the publication of the UK Government's Raptor Working Group Report that made recommendations for enhanced monitoring, ii) increased applied data needs (e.g., for site designation), and iii) concerns for the status of some species. The SRMS has 3 major objectives: i) to facilitate cooperation between parties; ii) to provide robust information on Scottish raptor populations by determining trends in numbers, range, survival, and productivity and understanding the causes of change; and iii) to maintain high and uniform standards for the collection, collation, auditing, and analysis of data and reporting of information. Data are collected for 19 species: 14 diurnal raptors, 4 owls, and 1 corvid, the Common Raven. Here we describe the development of the scheme, challenges, and achievements during its first 4 y, the nature and value of the data collected, and plans for the future.


Assuntos
Conservação dos Recursos Naturais , Monitoramento Ambiental/métodos , Falconiformes/crescimento & desenvolvimento , Animais , Conservação dos Recursos Naturais/legislação & jurisprudência , Conservação dos Recursos Naturais/métodos , Conservação dos Recursos Naturais/tendências , Monitoramento Ambiental/legislação & jurisprudência , Extinção Biológica , Programas Governamentais , Dinâmica Populacional , Escócia
15.
Biol Rev Camb Philos Soc ; 93(4): 1915-1937, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29790246

RESUMO

The impact of increasing vertebrate predator numbers on bird populations is widely debated among the general public, game managers and conservationists across Europe. However, there are few systematic reviews of whether predation limits the population sizes of European bird species. Views on the impacts of predation are particularly polarised in the UK, probably because the UK has a globally exceptional culture of intensive, high-yield gamebird management where predator removal is the norm. In addition, most apex predators have been exterminated or much depleted in numbers, contributing to a widely held perception that the UK has high numbers of mesopredators. This has resulted in many high-quality studies of mesopredator impacts over several decades. Here we present results from a systematic review of predator trends and abundance, and assess whether predation limits the population sizes of 90 bird species in the UK. Our results confirm that the generalist predators Red Fox (Vulpes vulpes) and Crows (Corvus corone and C. cornix) occur at high densities in the UK compared with other European countries. In addition, some avian and mammalian predators have increased numerically in the UK during recent decades. Despite these high and increasing densities of predators, we found little evidence that predation limits populations of pigeons, woodpeckers and passerines, whereas evidence suggests that ground-nesting seabirds, waders and gamebirds can be limited by predation. Using life-history characteristics of prey species, we found that mainly long-lived species with high adult survival and late onset of breeding were limited by predation. Single-brooded species were also more likely to be limited by predation than multi-brooded species. Predators that depredate prey species during all life stages (i.e. from nest to adult stages) limited prey numbers more than predators that depredated only specific life stages (e.g. solely during the nest phase). The Red Fox and non-native mammals (e.g. the American Mink Neovison vison) were frequently identified as numerically limiting their prey species. Our review has identified predator-prey interactions that are particularly likely to result in population declines of prey species. In the short term, traditional predator-management techniques (e.g. lethal control or fencing to reduce predation by a small number of predator species) could be used to protect these vulnerable species. However, as these techniques are costly and time-consuming, we advocate that future research should identify land-use practices and landscape configurations that would reduce predator numbers and predation rates.


Assuntos
Aves/fisiologia , Mamíferos/fisiologia , Comportamento Predatório , Animais , Densidade Demográfica , Reino Unido
16.
Ecol Lett ; 10(1): 25-35, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17204114

RESUMO

A key assumption underlying any management practice implemented to aid wildlife conservation is that it will have similar effects on target species across the range it is applied. However, this basic assumption is rarely tested. We show that predictors [nearly all associated with agri-environment scheme (AES) options known to affect European birds] had similar effects for 11 bird species on sites with differing farming practice (pastoral vs. mixed farming) or which differed in the density at which the species was found. However, predictors from sites in one geographical region tended to have different effects in other areas suggesting that AES options targeted at a regional scale are more likely to yield beneficial results for farmland birds than options applied uniformly in national schemes. Our study has broad implications for designing conservation strategies at an appropriate scale, which we discuss.


Assuntos
Aves , Conservação dos Recursos Naturais , Ecossistema , Agricultura , Animais , Europa (Continente) , Geografia , Modelos Estatísticos
17.
J Biomed Opt ; 12(3): 030503, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17614706

RESUMO

Angularly resolved light scattering measurements made at visible wavelengths have the ability to quantify subcellular morphology, with particular sensitivity to organelles the size of mitochondria and lysosomes. We have recently reported on a lysosome-staining-based method that provides scattering contrast between stained and unstained cells, and through the use of appropriate models, we extracted a size distribution and contribution to cellular light scattering that we attributed to lysosomes. We provide an independent measurement of the lysosomal size distribution and contribution to cellular light scattering by exploiting photodynamic ablation of lysosomes and observing its effect on angularly resolved light scattering measurements. From these measurements, we conclude that lysosomes scatter approximately 14% of the light from EMT6 cells at 633 nm and that their size distribution has a mean and standard deviation of 0.8 and 0.4 microm, respectively.


Assuntos
Lisossomos/fisiologia , Microscopia de Fluorescência/métodos , Modelos Biológicos , Organelas/fisiologia , Células Cultivadas , Simulação por Computador , Luz , Tamanho da Partícula , Espalhamento de Radiação
18.
J Biomed Opt ; 12(1): 014010, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17343485

RESUMO

Angularly resolved light scattering and wavelength-resolved darkfield scattering spectroscopy measurements were performed on intact, control EMT6 cells and cells stained with high-extinction lysosomal- or mitochondrial-localizing dyes. In the presence of the lysosomal-localizing dye NPe6, we observe changes in the details of light scattering from stained and unstained cells, which have both wavelength- and angular-dependent features. Analysis of measurements performed at several wavelengths reveals a reduced scattering cross section near the absorption maximum of the lysosomal-localizing dye. When identical measurements are made with cells loaded with a similar mitochondrial-localizing dye, HPPH, we find no evidence that staining mitochondria had any effect on the light scattering. Changes in the scattering properties of candidate populations of organelles induced by the addition of an absorber are modeled with Mie theory, and we find that any absorber-induced scattering response is very sensitive to the inherent refractive index of the organelle population. Our measurements and modeling are consistent with EMT6-cell-mitochondria having refractive indices close to those reported in the literature for organelles, approximately 1.4. The reduction in scattering cross section induced by NPe6 constrains the refractive index of lysosomes to be significantly higher. We estimate the refractive index of lysosomes in EMT6 cells to be approximately 1.6.


Assuntos
Clorofila/análogos & derivados , Modelos Biológicos , Porfirinas , Refratometria/métodos , Sarcoma/fisiopatologia , Espectrometria de Fluorescência/métodos , Frações Subcelulares , Animais , Linhagem Celular Tumoral , Simulação por Computador , Lisossomos , Camundongos , Microscopia de Fluorescência/métodos , Mitocôndrias , Espalhamento de Radiação
19.
Photochem Photobiol ; 83(5): 1056-62, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17880500

RESUMO

Phthalocyanine 4 (Pc 4) is a promising photosensitizer currently in clinical trials. Photobiological responses to Pc 4 photodynamic therapy (Pc 4-PDT) have been characterized extensively, but relatively little has been done to evaluate dose metrics for this sensitizer. We describe an irradiation-induced increase in fluorescence in tumor cell monolayers. This increase is due solely to enhanced fluorescence from Pc 4, as confirmed by confocal spectroscopy. In EMT6 cells incubated with 250 nM Pc 4 for 24 h, the maximum increase in fluorescence is approximately 3.7-fold above baseline levels. This increase occurs over a range of fluences, 0.05-0.6 J cm(-2), where clonogenic survival decreases by 3 orders of magnitude. Light scattering measurements performed on similarly treated EMT6 cells in suspension suggested a Pc 4-PDT-mediated mitochondrial swelling of approximately 13% at 0.6 J cm(-2), where fluorescence enhancement saturates under these treatment conditions. Fluorescence imaging and light scattering experiments performed at a five-fold lower Pc 4 incubation concentration revealed a reduced fluorescence enhancement at a five-fold higher fluence, which produced comparable mitochondrial swelling. Taken together, these data suggest that Pc 4 is initially aggregated at high local concentration in mitochondria and that irradiation relaxes the quenching of Pc 4 fluorescence through a mechanism that may involve mitochondrial swelling.


Assuntos
Indóis/farmacologia , Animais , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Fluorescência , Luz , Camundongos , Espalhamento de Radiação
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