RESUMO
Mounting evidence suggests that climate change will cause shifts of tree species range and abundance (biomass). Abundance changes under climate change are likely to occur prior to a detectable range shift. Disturbances are expected to directly affect tree species abundance and composition, and could profoundly influence tree species spatial distribution within a geographical region. However, how multiple disturbance regimes will interact with changing climate to alter the spatial distribution of species abundance remains unclear. We simulated such forest demographic processes using a forest landscape succession and disturbance model (LANDIS-II) parameterized with forest inventory data in the northeastern United States. Our study incorporated climate change under a high-emission future and disturbance regimes varying with gradients of intensities and spatial extents. The results suggest that disturbances catalyze changes in tree species abundance and composition under a changing climate, but the effects of disturbances differ by intensity and extent. Moderate disturbances and large extent disturbances have limited effects, while high-intensity disturbances accelerate changes by removing cohorts of mid- and late-successional species, creating opportunities for early-successional species. High-intensity disturbances result in the northern movement of early-successional species and the southern movement of late-successional species abundances. Our study is among the first to systematically investigate how disturbance extent and intensity interact to determine the spatial distribution of changes in species abundance and forest composition.
Assuntos
Mudança Climática , Árvores , Biomassa , Florestas , New EnglandRESUMO
The nutrient sensor O-GlcNAc transferase (OGT) catalyzes posttranslational addition of O-GlcNAc onto target proteins, influencing signaling pathways in response to cellular nutrient levels. OGT is highly expressed in pancreatic glucagon-secreting cells (α-cells), which secrete glucagon in response to hypoglycemia. The objective of this study was to determine whether OGT is necessary for the regulation of α-cell mass and function in vivo. We utilized genetic manipulation to produce two α-cell specific OGT-knockout models: a constitutive glucagon-Cre (αOGTKO) and an inducible glucagon-Cre (i-αOGTKO), which effectively delete OGT in α-cells. Using approaches including immunoblotting, immunofluorescent imaging, and metabolic phenotyping in vivo, we provide the first insight on the role of O-GlcNAcylation in α-cell mass and function. αOGTKO mice demonstrated normal glucose tolerance and insulin sensitivity but displayed significantly lower glucagon levels during both fed and fasted states. αOGTKO mice exhibited significantly lower α-cell glucagon content and α-cell mass at 6 months of age. In fasting, αOGTKO mice showed impaired pyruvate stimulated gluconeogenesis in vivo and reduced glucagon secretion in vitro. i-αOGTKO mice showed similarly reduced blood glucagon levels, defective in vitro glucagon secretion, and normal α-cell mass. Interestingly, both αOGTKO and i-αOGTKO mice had no deficiency in maintaining blood glucose homeostasis under fed or fasting conditions, despite impairment in α-cell mass and function, and glucagon content. In conclusion, these studies provide a first look at the role of OGT signaling in the α-cell, its effect on α-cell mass, and its importance in regulating glucagon secretion in hypoglycemic conditions.
Assuntos
Glicemia/metabolismo , Células Secretoras de Glucagon/enzimologia , Glucagon/biossíntese , N-Acetilglucosaminiltransferases/genética , Obesidade/genética , Acilação/efeitos dos fármacos , Animais , Jejum/metabolismo , Feminino , Efeito Fundador , Glucagon/deficiência , Células Secretoras de Glucagon/efeitos dos fármacos , Células Secretoras de Glucagon/patologia , Gluconeogênese/efeitos dos fármacos , Gluconeogênese/genética , Teste de Tolerância a Glucose , Fator 3-beta Nuclear de Hepatócito/genética , Fator 3-beta Nuclear de Hepatócito/metabolismo , Resistência à Insulina , Integrases/genética , Integrases/metabolismo , Masculino , Camundongos , Camundongos Knockout , N-Acetilglucosaminiltransferases/deficiência , Obesidade/enzimologia , Obesidade/patologia , Ácido Pirúvico/metabolismo , Ácido Pirúvico/farmacologiaRESUMO
DNA amplification occurs at the DNA puff II/9A locus in the fungus fly Sciara coprophila. As a foundation to study the molecular mechanism for the initiating events of II/9A DNA re-replication, we have sequenced 14 kb spanning a DNase hypersensitive site (DHS) upstream of the 1 kb amplification origin and through transcription units II/9-1 and II/9-2 downstream of the origin. These elements are annotated as well as the ORC binding site at the origin and the transition point (TP) between continuous and discontinuous DNA syntheses that marks the origin of bidirectional replication at the nucleotide level. A 9 bp motif found at the TP is repeated near the other end of the 1 kb ORI and may identify a putative second TP. The steroid hormone ecdysone induces DNA amplification as well as transcription and puffing at locus II/9A. Within the 14 kb, several matches to the ecdysone response element (EcRE) consensus sequence were identified, including some in the amplification origin region. EcRE O-P is at a central axis of a remarkable symmetry, equidistant to the TPs that are themselves equidistant to EcRE O-1 and EcRE O-2. DNA sequence alterations have occurred throughout the II/9A region in a newly discovered polymorphism (#2). Polymorphism #2 is not specific to developmental stage, sex, or tissue, and it does not impair DNA amplification. The DHS, both 9 bp TP sequences, and EcREs O-1, O-P, and O-2 are conserved between the polymorphism #1 and #2 sequences, suggesting their functional importance and retention during evolutionary selection. Moreover, a 72 bp sequence in the Sciara DHS at DNA puff II/9A is conserved in DNA puff C-3 of Rhynchosciara americana. Comparisons are discussed between the Sciara II/9A amplicon and the chorion locus amplicon on the third chromosome of Drosophila.
Assuntos
Replicação do DNA , Origem de Replicação , Animais , DNA/genética , DNA/metabolismo , Drosophila/genética , Larva/metabolismoRESUMO
Amorphous tantala (Ta2O5) thin films were deposited by reactive ion beam sputtering with simultaneous low energy assist Ar+ or Ar+/O2+ bombardment. Under the conditions of the experiment, the as-deposited thin films are amorphous and stoichiometric. The refractive index and optical band gap of thin films remain unchanged by ion bombardment. Around 20% improvement in room temperature mechanical loss and 60% decrease in absorption loss are found in samples bombarded with 100-eV Ar+. A detrimental influence from low energy O2+ bombardment on absorption loss and mechanical loss is observed. Low energy Ar+ bombardment removes excess oxygen point defects, while O2+ bombardment introduces defects into the tantala films.
RESUMO
TAF4b is a gonadal-enriched subunit of the general transcription factor TFIID that is implicated in promoting healthy ovarian aging and female fertility in mice and humans. To further explore the potential mechanism of TAF4b in promoting ovarian follicle development, we analyzed global gene expression at multiple time points in the human fetal ovary. This computational analysis revealed coordinate expression of human TAF4B and critical regulators and effectors of meiosis I including SYCP3, YBX2, STAG3, and DAZL. To address the functional relevance of this analysis, we turned to the embryonic Taf4b-deficient mouse ovary where, for the first time, we demonstrate, severe deficits in prophase I progression as well as asynapsis in Taf4b-deficient oocytes. Accordingly, TAF4b occupies the proximal promoters of many essential meiosis and oogenesis regulators, including Stra8, Dazl, Figla, and Nobox, and is required for their proper expression. These data reveal a novel TAF4b function in regulating a meiotic gene expression program in early mouse oogenesis, and support the existence of a highly conserved TAF4b-dependent gene regulatory network promoting early oocyte development in both mice and women.
Assuntos
Meiose/genética , Oócitos/metabolismo , Fatores Associados à Proteína de Ligação a TATA/genética , Fator de Transcrição TFIID/genética , Animais , Feminino , Expressão Gênica/genética , Redes Reguladoras de Genes/genética , Humanos , Masculino , Camundongos , Oogênese/genética , Ovário/metabolismo , Regiões Promotoras Genéticas/genéticaRESUMO
Climate change is expected to cause geographic shifts in tree species' ranges, but such shifts may not keep pace with climate changes because seed dispersal distances are often limited and competition-induced changes in community composition can be relatively slow. Disturbances may speed changes in community composition, but the interactions among climate change, disturbance and competitive interactions to produce range shifts are poorly understood. We used a physiologically based mechanistic landscape model to study these interactions in the northeastern United States. We designed a series of disturbance scenarios to represent varied disturbance regimes in terms of both disturbance extent and intensity. We simulated forest succession by incorporating climate change under a high-emissions future, disturbances, seed dispersal, and competition using the landscape model parameterized with forest inventory data. Tree species range boundary shifts in the next century were quantified as the change in the location of the 5th (the trailing edge) and 95th (the leading edge) percentiles of the spatial distribution of simulated species. Simulated tree species range boundary shifts in New England over the next century were far below (usually <20 km) that required to track the velocity of temperature change (usually more than 110 km over 100 years) under a high-emissions scenario. Simulated species` ranges shifted northward at both the leading edge (northern boundary) and trailing edge (southern boundary). Disturbances may expedite species' recruitment into new sites, but they had little effect on the velocity of simulated range boundary shifts. Range shifts at the trailing edge tended to be associated with photosynthetic capacity, competitive ability for light and seed dispersal ability, whereas shifts at the leading edge were associated only with photosynthetic capacity and competition for light. This study underscores the importance of understanding the role of interspecific competition and disturbance when studying tree range shifts.
Assuntos
Mudança Climática , Florestas , Árvores/fisiologia , Demografia , New England , SementesRESUMO
American chestnut (Castanea dentata) was once an important component forests in the central Appalachians (USA), but it was functionally extirpated nearly a century ago. Attempts are underway to reintroduce blight-resistant chestnut to its former range, but it is uncertain how current forest composition, climate, and atmospheric changes and disturbance regimes will interact to determine future forest dynamics and ecosystem services. The combination of novel environmental conditions (e.g. climate change), a reintroduced tree species and new disturbance regimes (e.g. exotic insect pests, fire suppression) have no analog in the past that can be used to parameterize phenomenological models. We therefore used a mechanistic approach within the LANDIS-II forest landscape model that relies on physiological first principles to project forest dynamics as the outcome of competition of tree cohorts for light and water as a function of temperature, precipitation, CO2 concentration, and life history traits. We conducted a factorial landscape simulation experiment to evaluate specific hypotheses about future forest dynamics in two study sites in the center of the former range of chestnut. Our results supported the hypotheses that climate change would favor chestnut because of its optimal temperature range and relative drought resistance, and that chestnut would be less competitive in the more mesic Appalachian Plateau province because competitors will be less stressed. The hypothesis that chestnut will increase carbon stocks was supported, although the increase was modest. Our results confirm that aggressive restoration is needed regardless of climate and soils, and that increased aggressiveness of chestnut restoration increased biomass accumulation. The hypothesis that chestnut restoration will increase both compositional and structural richness was not supported because chestnut displaced some species and age cohorts. Although chestnut restoration did not markedly enhance carbon stocks, our findings provide hope that this formerly important species can be successfully reintroduced and associated ecosystem services recovered.
Assuntos
Sequestro de Carbono , Mudança Climática , Conservação dos Recursos Naturais/métodos , Fagaceae/fisiologia , Árvores/fisiologia , Fagaceae/crescimento & desenvolvimento , Maryland , Árvores/crescimento & desenvolvimentoRESUMO
Malreduction of distal tibiofibular syndesmosis (DTFS) leads to poor functional outcomes after ankle fracture surgery. Difficulty achieving anatomic alignment of the syndesmosis is due to variable morphology of the fibular incisura of the tibia and a paucity of literature regarding its morphologic characteristics. We surveyed 775 consecutive ankle computed tomography (CT) scans performed from June 2008 to December 2011, and 203 (26.2%) were included for evaluation. Two observers performed quantitative measurements and qualitative evaluated fibular incisura morphology. Tang ratios for fibular rotation, anterior and posterior tibiofibular distances, fibular incisura depth, and subjective morphologies on CT were assessed using conventional multiplanar reconstruction (MPR) and maximum intensity projections (MIPs). On conventional CT, the mean Tang ratio was 0.97 ± 0.06; the mean anterior tibiofibular distance was 2.17 ± 0.87 mm; the mean posterior tibiofibular distance was 3.52 ± 0.94 mm; and the mean depth of fibular incisura was 3.29 ± 1.19 mm. Five morphologic variations of the fibular incisura were identified: crescentic, trapezoid, flat, chevron, and widow's peak. The most common fibular incisura morphology was crescentic (61.3%), followed by trapezoid shape (25.1%); the least common morphology was flat (3.1%). Interobserver variability with intraclass correlation coefficient (ICC) was slightly higher for all quantitative measures on MPR (ICCâ¯=â¯0.72 to .81) versus MIP (ICCâ¯=â¯0.64 to 0.75). ICC for incisura shape and depth assessments was poor on both modalities (0.13 to 0.38). This comprehensive CT study reports on quantitative and qualitative descriptive measures to evaluate fibular incisura morphologies and fibular orientation. It also defines the frequency of DTFS measures and the interobserver performance on 2 CT evaluation methods.
Assuntos
Articulação do Tornozelo/anatomia & histologia , Articulação do Tornozelo/diagnóstico por imagem , Imageamento Tridimensional , Tomografia Computadorizada por Raios X , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Valores de Referência , Estudos RetrospectivosRESUMO
Long-term mammalian spermatogenesis requires proper development of spermatogonial stem cells (SSCs) that replenish the testis with germ cell progenitors during adult life. TAF4b is a gonadal-enriched component of the general transcription factor complex, TFIID, which is required for the maintenance of spermatogenesis in the mouse. Successful germ cell transplantation assays into adult TAF4b-deficient host testes suggested that TAF4b performs an essential germ cell autonomous function in SSC establishment and/or maintenance. To elucidate the SSC function of TAF4b, we characterized the initial gonocyte pool and rounds of spermatogenic differentiation in the context of the Taf4b-deficient mouse testis. Here, we demonstrate a significant reduction in the late embryonic gonocyte pool and a deficient expansion of this pool soon after birth. Resulting from this reduction of germ cell progenitors is a developmental delay in meiosis initiation, as compared to age-matched controls. While GFRα1+ spermatogonia are appropriately present as Asingle and Apaired in wild-type testes, TAF4b-deficient testes display an increased proportion of long and clustered chains of GFRα1+ cells. In the absence of TAF4b, seminiferous tubules in the adult testis either lack germ cells altogether or are found to have missing generations of spermatogenic progenitor cells. Together these data indicate that TAF4b-deficient spermatogenic progenitor cells display a tendency for differentiation at the expense of self-renewal and a renewing pool of SSCs fail to establish during the critical window of SSC development.
Assuntos
Células-Tronco Adultas/fisiologia , Diferenciação Celular/fisiologia , Espermatogênese/fisiologia , Espermatogônias/crescimento & desenvolvimento , Fatores Associados à Proteína de Ligação a TATA/biossíntese , Fator de Transcrição TFIID/biossíntese , Animais , Animais Recém-Nascidos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos KnockoutRESUMO
This is an overview of the adaptive optics used in Advanced LIGO (aLIGO), known as the thermal compensation system (TCS). The TCS was designed to minimize thermally induced spatial distortions in the interferometer optical modes and to provide some correction for static curvature errors in the core optics of aLIGO. The TCS is comprised of ring heater actuators, spatially tunable CO2 laser projectors, and Hartmann wavefront sensors. The system meets the requirements of correcting for nominal distortion in aLIGO to a maximum residual error of 5.4 nm rms, weighted across the laser beam, for up to 125 W of laser input power into the interferometer.
RESUMO
KEY POINTS: Activation of NMDA receptors (NMDARs) is essential for encoding visual stimuli into signals for the brain, although their over-activation can cause cell death. The recruitment of NMDARs is important for encoding light intensity in retinal ganglion cells. D-serine binding is essential for proper activation of NMDARs, although its role in signal processing and the mechanisms that underlie its availability are not well understood. In these light-evoked experiments, the addition of exogenous D-serine had a large effect on low contrast and low intensity NMDAR responses that decreased as the intensity was increased. The degradation of endogenous D-serine decreased the responses more at higher intensities. The results provide compelling evidence favouring a new interpretation of NMDAR recruitment in which light-evoked D-serine release serves an important regulatory control over the recruitment of NMDARs. ABSTRACT: The present study aimed to investigate the functional properties of NMDA receptor coagonist release and to specifically evaluate whether light-evoked release mechanisms contribute to the availability of the coagonist D-serine. Two different methods were involved in our approach: (i) whole-cell recordings from identified retinal ganglion cells in the tiger salamander were used to study light adaptation with positive and negative contrast stimuli over a range of ± 1 log unit against a steady background illumination and (ii) the mechanisms for intensity encoding to a range of light intensities covering 6 log10 units were investigated. This latter study employed extracellular recordings of the proximal negative field potential, pharmacologically manipulated to generate a pure NMDA mediated response. For the adaptation study, we examined the light-evoked responses under control conditions, followed by light stimuli presented in the presence of D-serine, followed by light stimulation in the presence of dichlorokynurenic acid to block the coagonist site of NMDA receptors. For the brightness encoding studies, we examined the action of D-serine on each intensity used and then applied the enzyme D-serine deaminase to remove significant levels of D-serine. These studies provided new insights into the mechanisms that regulate coagonist availability in the vertebrate retina. Our results strongly support the idea that light-evoked coagonist release, a major component of which is D-serine, is needed to provide the full range of coagonist availability for optimal activation of NMDA receptors.
Assuntos
Receptores de N-Metil-D-Aspartato/fisiologia , Células Ganglionares da Retina/fisiologia , Serina/fisiologia , Ambystoma , Animais , Estimulação LuminosaRESUMO
Fundamental drivers of ecosystem processes such as temperature and precipitation are rapidly changing and creating novel environmental conditions. Forest landscape models (FLM) are used by managers and policy-makers to make projections of future ecosystem dynamics under alternative management or policy options, but the links between the fundamental drivers and projected responses are weak and indirect, limiting their reliability for projecting the impacts of climate change. We developed and tested a relatively mechanistic method to simulate the effects of changing precipitation on species competition within the LANDIS-II FLM. Using data from a field precipitation manipulation experiment in a piñon pine (Pinus edulis) and juniper (Juniperus monosperma) ecosystem in New Mexico (USA), we calibrated our model to measurements from ambient control plots and tested predictions under the drought and irrigation treatments against empirical measurements. The model successfully predicted behavior of physiological variables under the treatments. Discrepancies between model output and empirical data occurred when the monthly time step of the model failed to capture the short-term dynamics of the ecosystem as recorded by instantaneous field measurements. We applied the model to heuristically assess the effect of alternative climate scenarios on the piñon-juniper ecosystem and found that warmer and drier climate reduced productivity and increased the risk of drought-induced mortality, especially for piñon. We concluded that the direct links between fundamental drivers and growth rates in our model hold great promise to improve our understanding of ecosystem processes under climate change and improve management decisions because of its greater reliance on first principles.
Assuntos
Mudança Climática , Secas , Florestas , Juniperus/fisiologia , Pinus/fisiologia , Carbono/metabolismo , Modelos Teóricos , Fotossíntese , Transpiração VegetalRESUMO
BACKGROUND: Piwi proteins are essential for germ line development, stem cell maintenance, and more recently found to function in epigenetic and somatic gene regulation. In the sea urchin Strongylocentrotus purpuratus, two Piwi proteins, Seawi and Piwi-like1, have been identified, yet their functional contributions have not been reported. RESULTS: Here we found that Seawi protein was localized uniformly in the early embryo and then became enriched in the primordial germ cells (PGCs) (the small micromere lineage) from blastula stage and thereafter. Morpholino knockdown of Sp-seawi diminished PGC-specific localization of Seawi proteins, and altered expression of other germ line markers such as Vasa and Gustavus, but had no effect on Nanos. Furthermore, Seawi knockdown transiently resulted in Vasa positive cell proliferation in the right coelomic pouch that appear to be derived from the small micromere lineage, yet they quickly disappeared with an indication of apoptosis by larval stage. Severe Seawi knockdown resulted in an increased number of apoptotic cells in the entire gut area. CONCLUSION: Piwi proteins appear to regulate PGC proliferation perhaps through control of Vasa accumulation. In this organism, Piwi is likely regulating mRNAs, not just transposons, and is potentially functioning both inside and outside of the germ line during embryogenesis.
Assuntos
Proteínas Argonautas/metabolismo , RNA Helicases DEAD-box/metabolismo , Desenvolvimento Embrionário/fisiologia , Strongylocentrotus purpuratus/embriologia , Animais , Apoptose/fisiologia , Proteínas Argonautas/genética , Proliferação de Células , RNA Helicases DEAD-box/genética , Técnicas de Silenciamento de Genes , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Strongylocentrotus purpuratus/genéticaRESUMO
The formation of the germ line in an embryo marks a fresh round of reproductive potential. The developmental stage and location within the embryo where the primordial germ cells (PGCs) form, however, differs markedly among species. In many animals, the germ line is formed by an inherited mechanism, in which molecules made and selectively partitioned within the oocyte drive the early development of cells that acquire this material to a germ-line fate. In contrast, the germ line of other animals is fated by an inductive mechanism that involves signaling between cells that directs this specialized fate. In this review, we explore the mechanisms of germ-line determination in echinoderms, an early-branching sister group to the chordates. One member of the phylum, sea urchins, appears to use an inherited mechanism of germ-line formation, whereas their relatives, the sea stars, appear to use an inductive mechanism. We first integrate the experimental results currently available for germ-line determination in the sea urchin, for which considerable new information is available, and then broaden the investigation to the lesser-known mechanisms in sea stars and other echinoderms. Even with this limited insight, it appears that sea stars, and perhaps the majority of the echinoderm taxon, rely on inductive mechanisms for germ-line fate determination. This enables a strongly contrasted picture for germ-line determination in this phylum, but one for which transitions between different modes of germ-line determination might now be experimentally addressed.
Assuntos
Equinodermos/embriologia , Indução Embrionária/fisiologia , Gametogênese/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Células Germinativas/fisiologia , Modelos Biológicos , Transdução de Sinais/fisiologia , Animais , Divisão Celular Assimétrica/fisiologia , Cálcio/metabolismo , Canais de Cálcio Tipo L/metabolismo , RNA Helicases DEAD-box/metabolismo , Retículo Endoplasmático/metabolismo , Especificidade da EspécieRESUMO
BACKGROUND: Meiosis is a unique mechanism in gamete production and a fundamental process shared by all sexually reproducing eukaryotes. Meiosis requires several specialized and highly conserved genes whose expression can also identify the germ cells undergoing gametogenic differentiation. Sea urchins are echinoderms, which form a phylogenetic sister group of chordates. Sea urchin embryos undergo a feeding, planktonic larval phase in which they construct an adult rudiment prior to metamorphosis. Although a series of conserved meiosis genes (e.g., dmc1, msh5, rad21, rad51, and sycp1) is expressed in sea urchin oocytes, we sought to determine when in development meiosis would first be initiated. RESULTS: We surveyed the expression of several meiotic genes and their corresponding proteins in the sea urchin Strongylocentrotus purpuratus. Surprisingly, meiotic genes are highly expressed not only in ovaries but beginning in larvae. Both RNA and protein localizations strongly suggest that meiotic gene expression initiates in tissues that will eventually give rise to the adult rudiment of the late larva. CONCLUSIONS: These results demonstrate that broad expression of the molecules associated with meiotic differentiation initiates prior to metamorphosis and may have additional functions in these cells, or mechanisms repressing their function, until later in development when gametogenesis begins.
Assuntos
Proteínas de Ciclo Celular/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Meiose/genética , Strongylocentrotus purpuratus/metabolismo , Animais , Sequência de Bases , Clonagem Molecular , Primers do DNA/genética , Perfilação da Expressão Gênica , Hibridização in Situ Fluorescente , Larva/crescimento & desenvolvimento , Larva/metabolismo , Meiose/fisiologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Strongylocentrotus purpuratus/crescimento & desenvolvimentoRESUMO
D-serine is present in the vertebrate retina and serves as a coagonist for the N-methyl-D-aspartate (NMDA) receptors of ganglion cells. Although the enzyme D-amino acid oxidase (DAO) has been implicated as a pathway for d-serine degradation, its role in the retina has not been established. In this study, we investigated the role of DAO in regulating D-serine levels using a mutant mouse line deficient in DAO (ddY/DAO(-)) and compared these results with their wild-type counterparts (ddY/DAO(+)). Our results show that DAO is functionally present in the mouse retina and normally serves to reduce the background levels of D-serine. The enzymatic activity of DAO was restricted to the inner plexiform layer as determined by histochemical analysis. Using capillary electrophoresis, we showed that mutant mice had much higher levels of D-serine. Whole cell recordings from identified retinal ganglion cells demonstrated that DAO-deficient animals had light-evoked synaptic activity strongly biased toward a high NMDA-to-AMPA receptor ratio. In contrast, recordings from wild-type ganglion cells showed a more balanced ratio between the two receptor subclasses. Immunostaining for AMPA and NMDA receptors was carried out to compare the two receptor ratios by quantitative immunofluorescence. These studies revealed that the mutant mouse had a significantly higher representation of NMDA receptors compared with the wild-type controls. We conclude that 1) DAO is an important regulatory enzyme and normally functions to reduce D-serine levels in the retina, and 2) D-serine levels play a role in the expression of NMDA receptors and the NMDA-to-AMPA receptor ratio.
Assuntos
D-Aminoácido Oxidase/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Retina/metabolismo , Potenciais de Ação , Animais , D-Aminoácido Oxidase/deficiência , Potenciais Pós-Sinápticos Excitadores , Camundongos , Mutação , Receptores de AMPA/genética , Receptores de AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/genética , Retina/enzimologia , Retina/fisiologia , Células Ganglionares da Retina/metabolismo , Células Ganglionares da Retina/fisiologia , Serina/química , Serina/metabolismo , EstereoisomerismoRESUMO
The importance of sexual dimorphism has been highlighted in recent years since the National Institutes of Health's mandate on considering sex as a biological variable. Although recent studies have taken strides to study both sexes side by side, investigations into the normal physiological differences between males and females are limited. In this study, we aimed to characterized sex-dependent differences in glucose metabolism and pancreatic ß-cell physiology in normal conditions using C57BL/6J mice, the most common mouse strain used in metabolic studies. Here, we report that female mice have improved glucose and insulin tolerance associated with lower nonfasted blood glucose and insulin levels compared with male mice at 3 and 6 months of age. Both male and female animals show ß-cell mass expansion from embryonic day 17.5 to adulthood, and no sex differences were observed at embryonic day 17.5, newborn, 1 month, or 3 months of age. However, 6-month-old males displayed increased ß-cell mass in response to insulin resistance compared with littermate females. Molecularly, we uncovered sexual dimorphic alterations in the protein levels of nutrient sensing proteins O-GlcNAc transferase and mTOR, as well as differences in glucose-stimulus coupling mechanisms that may underlie the differences in sexually dimorphic ß-cell physiology observed in C57BL/6J mice.
RESUMO
Vasa is a broadly conserved DEAD-box RNA helicase associated with germ line development and is expressed in multipotent cells in many animals. During embryonic development of the sea urchin Strongylocentrotus purpuratus, Vasa protein is enriched in the small micromeres despite a uniform distribution of vasa transcript. Here we show that the Vasa coding region is sufficient for its selective enrichment and find that gustavus, the B30.2/SPRY and SOCS box domain gene, contributes to this phenomenon. In vitro binding analyses show that Gustavus binds the N-terminal and DEAD-box portions of Vasa protein independently. A knockdown of Gustavus protein reduces both Vasa protein abundance and its propensity for accumulation in the small micromeres, whereas overexpression of the Vasa-interacting domain of Gustavus (GusΔSOCS) results in Vasa protein accumulation throughout the embryo. We propose that Gustavus has a conserved, positive regulatory role in Vasa protein accumulation during embryonic development.
Assuntos
RNA Helicases DEAD-box/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Células-Tronco Multipotentes/metabolismo , Strongylocentrotus purpuratus/embriologia , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Animais , Clonagem Molecular , Imunofluorescência , Imuno-Histoquímica , Hibridização In Situ , Mutagênese , Oligonucleotídeos , Filogenia , Strongylocentrotus purpuratus/metabolismo , Proteínas Supressoras da Sinalização de Citocina/genéticaRESUMO
Insect disturbance is often thought to increase fire risk through enhanced fuel loadings, particularly in coniferous forest ecosystems. Yet insect disturbances also affect successional pathways and landscape structure that interact with fire disturbances (and vice-versa) over longer time scales. We applied a landscape succession and disturbance model (LANDIS-II) to evaluate the relative strength of interactions between spruce budworm (Choristoneura fumiferana) outbreaks and fire disturbances in the Boundary Waters Canoe Area (BWCA) in northern Minnesota (USA). Disturbance interactions were evaluated for two different scenarios: presettlement forests and fire regimes vs. contemporary forests and fire regimes. Forest composition under the contemporary scenario trended toward mixtures of deciduous species (primarily Betula papyrifera and Populus spp.) and shade-tolerant conifers (Picea mariana, Abies balsamea, Thuja occidentalis), with disturbances dominated by a combination of budworm defoliation and high-severity fires. The presettlement scenario retained comparatively more "big pines" (i.e., Pinus strobus, P. resinosa) and tamarack (L. laricina), and experienced less budworm disturbance and a comparatively less-severe fire regime. Spruce budworm disturbance decreased area burned and fire severity under both scenarios when averaged across the entire 300-year simulations. Contrary to past research, area burned and fire severity during outbreak decades were each similar to that observed in non-outbreak decades. Our analyses suggest budworm disturbances within forests of the BWCA have a comparatively weak effect on long-term forest composition due to a combination of characteristics. These include strict host specificity, fine-scaled patchiness created by defoliation damage, and advance regeneration of its primary host, balsam fir (A. balsamea) that allows its host to persist despite repeated disturbances. Understanding the nature of the three-way interaction between budworm, fire, and composition has important ramifications for both fire mitigation strategies and ecosystem restoration initiatives. We conclude that budworm disturbance can partially mitigate long-term future fire risk by periodically reducing live ladder fuel within the mixed forest types of the BWCA but will do little to reverse the compositional trends caused in part by reduced fire rotations.
Assuntos
Ecossistema , Incêndios , Mariposas/fisiologia , Árvores , Animais , Simulação por Computador , Conservação dos Recursos Naturais , Monitoramento Ambiental , Comportamento Alimentar , Larva/fisiologia , Minnesota , Modelos Biológicos , Dinâmica Populacional , Fatores de TempoRESUMO
Sexually reproducing metazoans establish a cell lineage during development that is ultimately dedicated to gamete production. Work in a variety of animals suggests that a group of conserved molecular determinants act in this germ line maintenance and function. The most universal of these genes are Vasa and Vasa-like DEAD-box RNA helicase genes. However, recent evidence indicates that Vasa genes also function in other cell types, distinct from the germ line. Here we evaluate our current understanding of Vasa function and its regulation during development, addressing Vasa's emerging role in multipotent cells. We also explore the evolutionary diversification of the N-terminal domain of this gene and how this impacts the association of Vasa with nuage-like perinuclear structures.