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1.
Nat Commun ; 15(1): 3453, 2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38658623

RESUMO

Carbon removal is emerging as a pillar of governmental and industry commitments toward achieving Net Zero targets. Drawing from 44 focus groups in 22 countries, we map technical and societal issues that a representative sample of publics raise on five major types of carbon removal (forests, soils, direct air capture, enhanced weathering, and bioenergy with carbon capture and storage), and how these translate to preferences for governance actors, mechanisms, and rationales. We assess gaps and overlaps between a global range of public perceptions and how carbon removal is currently emerging in assessment, innovation, and decision-making. In conclusion, we outline key societal expectations for informing assessment and policy: prioritize public engagement as more than acceptance research; scrutiny and regulation of industry beyond incentivizing innovation; systemic coordination across sectors, levels, and borders; and prioritize underlying causes of climate change and interrelated governance issues.


Assuntos
Carbono , Mudança Climática , Grupos Focais , Opinião Pública , Humanos , Solo/química , Sequestro de Carbono , Feminino , Masculino , Adulto
2.
Env Polit ; 33(2): 340-365, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38444630

RESUMO

Institutional theory, behavioral science, sociology and even political science all emphasize the importance of actors in achieving social change. Despite this salience, the actors involved in researching, promoting, or deploying negative emissions and solar geoengineering technologies remain underexplored within the literature. In this study, based on a rigorous sample of semi-structured expert interviews (N = 125), we empirically explore the types of actors and groups associated with both negative emissions and solar geoengineering research and deployment. We investigate emergent knowledge networks and patterns of involvement across space and scale. We examine actors in terms of their support of, opposition to, or ambiguity regarding both types of climate interventions. We reveal incipient and perhaps unforeseen collections of actors; determine which sorts of actors are associated with different technology pathways to comprehend the locations of actor groups and potential patterns of elitism; and assess relative degrees of social acceptance, legitimacy, and governance.

3.
Nat Commun ; 15(1): 2060, 2024 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-38448460

RESUMO

Novel, potentially radical climate intervention technologies like carbon dioxide removal and solar geoengineering are attracting attention as the adverse impacts of climate change are increasingly felt. The ability of publics, particularly in the Global South, to participate in discussions about research, policy, and deployment is restricted amidst a lack of familiarity and engagement. Drawing on a large-scale, cross-country exercise of nationally representative surveys (N = 30,284) in 30 countries and 19 languages, this article establishes the first global baseline of public perceptions of climate-intervention technologies. Here, we show that Global South publics are significantly more favorable about potential benefits and express greater support for climate-intervention technologies. The younger age and level of climate urgency and vulnerability of these publics emerge as key explanatory variables, particularly for solar geoengineering. Conversely, Global South publics express greater concern that climate-intervention technologies could undermine climate-mitigation efforts, and that solar geoengineering could promote an unequal distribution of risks between poor and rich countries.

4.
Glob Environ Change ; 83: 102765, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38130391

RESUMO

Public perception of emerging climate technologies, such as greenhouse gas removal (GGR) and solar radiation management (SRM), will strongly influence their future development and deployment. Studying perceptions of these technologies with traditional survey methods is challenging, because they are largely unknown to the public. Social media data provides a complementary line of evidence by allowing for retrospective analysis of how individuals share their unsolicited opinions. Our large-scale, comparative study of 1.5 million tweets covers 16 GGR and SRM technologies and uses state-of-the-art deep learning models to show how attention, and expressions of sentiment and emotion developed between 2006 and 2021. We find that in recent years, attention has shifted from general geoengineering themes to specific GGR methods. On the other hand, there is little attention to specific SRM technologies and they often coincide with conspiracy narratives. Sentiments and emotions in GGR tweets tend to be more positive, particularly for methods perceived to be natural, but are more negative when framed in the geoengineering context.

5.
Risk Anal ; 43(4): 838-859, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35508324

RESUMO

Deliberations are underway to utilize increasingly radical technological options to help address climate change and stabilize the climatic system. Collectively, these options are often referred to as "climate geoengineering." Deployment of such options, however, can create wicked tradeoffs in governance and require adaptive forms of risk management. In this study, we utilize a large and novel set of qualitative expert interview data to more deeply and systematically explore the types of risk-risk tradeoffs that may emerge from the use of 20 different climate geoengineering options, 10 that focus on carbon dioxide or greenhouse gas removal, and 10 that focus on solar radiation management and reflecting sunlight. We specifically consider: What risks does the deployment of these options entail? What types of tradeoffs may emerge through their deployment? We apply a framework that clusters risk-risk tradeoffs into institutional and governance, technological and environmental, and behavioral and temporal dimensions. In doing so, we offer a more complete inventory of risk-risk tradeoffs than those currently available within the respective risk-assessment, energy-systems, and climate-change literatures, and we also point the way toward future research gaps concerning policy, deployment, and risk management.

6.
Sci Prog ; 105(4): 368504221138443, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36476205

RESUMO

This article discusses and illuminates the synergies and jeopardies or tradeoffs that exist between the 17 Sustainable Development Goals (SDGs) and net-zero or future climate protection options such as greenhouse gas removal (GGR) technologies and solar radiation management (SRM) deployment approaches, respectively. Through a large-scale expert-interview exercise (N = 125), the study finds firstly that numerous synergies and tradeoffs exist between GGR, SRM, and the SDGs. More specifically, we reveal that GGR deployment could enhance the attainment of 16 of the 17 SDGs, but this comes with possible tradeoffs with 12 of the SDGs. SRM deployment could not only enhance the attainment of 16 of the 17 SDGs, but also create possible tradeoffs with (a different) 12 SDGs. The findings further support the understanding of the complexity of SRM and GGR proposals and help policymakers and industrial pioneers understand, navigate, and benchmark between geoengineering approaches using sustainable development goals.


Assuntos
Exercício Físico , Desenvolvimento Sustentável
7.
Artigo em Inglês | MEDLINE | ID: mdl-36200076

RESUMO

Negative emissions technologies and solar radiation management techniques could contribute towards climate stability, either by removing carbon dioxide from the atmosphere and storing it permanently or reflecting sunlight away from the atmosphere. Despite concerns about them, such options are increasingly being discussed as crucial complements to traditional climate change mitigation and adaptation. Expectations around negative emissions and solar radiation management and their associated risks and costs shape public and private discussions of how society deals with the climate crisis. In this study, we rely on a large expert survey (N = 74) to critically examine the future potential of both negative emission options (e.g., carbon dioxide removal) and solar radiation management techniques. We designed a survey process that asked a pool of prominent experts questions about (i) the necessity of adopting negative emissions or solar radiation management options, (ii) the desirability of such options when ranked against each other, (iii) estimations of future efficacy in terms of temperature reductions achieved or gigatons of carbon removed, (iv) expectations about future scaling, commercialization, and deployment targets, and (v) potential risks and barriers. Unlike other elicitation processes where experts are more positive or have high expectations about novel options, our results are more critical and cautionary. We find that some options (notably afforestation and reforestation, ecosystem restoration, and soil carbon sequestration) are envisioned frequently as necessary, desirable, feasible, and affordable, with minimal risks and barriers (compared to other options). This contrasts with other options envisaged as unnecessary risky or costly, notably ocean alkalization or fertilization, space-based reflectors, high-altitude sunshades, and albedo management via clouds. Moreover, only the options of afforestation and reforestation and soil carbon sequestration are expected to be widely deployed before 2035, which raise very real concerns about climate and energy policy in the near- to mid-term.

8.
Risk Anal ; 42(9): 1965-1979, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-32722874

RESUMO

Climate change is a paradigmatic example of systemic risk. Recently, proposals for large-scale interventions-carbon dioxide removal (CDR) and solar radiation management (SRM)-have started to redefine climate governance strategies. We describe how evolving modeling practices are trending toward optimized and "best-case" projections-portraying deployment schemes that create both technically slanted and politically sanitized profiles of risk, as well as ideal objectives for CDR and SRM as mitigation-enhancing, time-buying mechanisms for carbon transitions or vulnerable populations. As promises, stylized and hopeful projections may selectively reinforce industry and political activities built around the inertia of the carbon economy. Some evidence suggests this is the emerging case for certain kinds of CDR, where the prospect of future carbon capture substitutes for present mitigation. Either of these implications are systemic: explorations of climatic futures may entrench certain carbon infrastructures. We point out efforts and recommendations to forestall this trend in the implementation of the Paris Agreement, by creating more stakeholder input and strengthening political realism in modeling and other assessments, as well as through policy guardrails.


Assuntos
Dióxido de Carbono , Luz Solar , Dióxido de Carbono/química , Mudança Climática , Previsões
9.
Environ Sci Technol ; 55(17): 11925-11936, 2021 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-34291911

RESUMO

Biomass waste-derived porous carbons (BWDPCs) are a class of complex materials that are widely used in sustainable waste management and carbon capture. However, their diverse textural properties, the presence of various functional groups, and the varied temperatures and pressures to which they are subjected during CO2 adsorption make it challenging to understand the underlying mechanism of CO2 adsorption. Here, we compiled a data set including 527 data points collected from peer-reviewed publications and applied machine learning to systematically map CO2 adsorption as a function of the textural and compositional properties of BWDPCs and adsorption parameters. Various tree-based models were devised, where the gradient boosting decision trees (GBDTs) had the best predictive performance with R2 of 0.98 and 0.84 on the training and test data, respectively. Further, the BWDPCs in the compiled data set were classified into regular porous carbons (RPCs) and heteroatom-doped porous carbons (HDPCs), where again the GBDT model had R2 of 0.99 and 0.98 on the training and 0.86 and 0.79 on the test data for the RPCs and HDPCs, respectively. Feature importance revealed the significance of adsorption parameters, textural properties, and compositional properties in the order of precedence for BWDPC-based CO2 adsorption, effectively guiding the synthesis of porous carbons for CO2 adsorption applications.


Assuntos
Dióxido de Carbono , Carbono , Adsorção , Biomassa , Aprendizado de Máquina , Porosidade
10.
Front Mol Neurosci ; 11: 286, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30323738

RESUMO

To study characterization of zebrafish glycine receptors (zGlyRs), we assessed expression and function of five α- and two ß-subunit encoding GlyR in zebrafish. Our qPCR analysis revealed variable expression during development, while in situ hybridizations uncovered expression in the hindbrain and spinal cord; a finding consistent with the reported expression of GlyR subunits in these tissues from other organisms. Electrophysiological recordings using Xenopus oocytes revealed that all five α subunits form homomeric receptors activated by glycine, and inhibited by strychnine and picrotoxin. In contrast, ß subunits only formed functional heteromeric receptors when co-expressed with α subunits. Curiously, the second transmembranes of both ß subunits were found to lack a phenylalanine at the sixth position that is commonly associated with conferring picrotoxin resistance to heteromeric receptors. Consistent with the absence of phenylalanines at the sixth position, heteromeric zGlyRs often lacked significant picrotoxin resistance. Subsequent efforts revealed that resistance to picrotoxin in both zebrafish and human heteromeric GlyRs involves known residues within transmembrane 2, as well as previously unknown residues within transmembrane 3. We also found that a dominant mutation in human GlyRα1 that gives rise to hyperekplexia, and recessive mutations in zebrafish GlyRßb that underlie the bandoneon family of motor mutants, result in reduced receptor function. Lastly, through the use of a concatenated construct we demonstrate that zebrafish heteromeric receptors assemble with a stoichiometry of 3α:2ß. Collectively, our findings have furthered our knowledge regarding the assembly of heteromeric receptors, and the molecular basis of ß subunit-conferred picrotoxin resistance. These results should aid in future investigations of glycinergic signaling in zebrafish and mammals.

11.
Sci Total Environ ; 580: 90-104, 2017 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-28011016

RESUMO

Since solar radiation management (SRM) technologies do not yet exist and capacities to model their impacts are limited, proposals for its governance are implicitly designed not around realities, but possibilities - baskets of risk and benefit that are often components of future imaginaries. This paper reports on the project Solar Radiation Management: Foresight for Governance (SRM4G), which aimed to encourage an anticipatory mode of thinking about the future of an engineered climate. Leveraging the participation of 15 scholars and practitioners heavily engaged in early conversations on SRM governance, SRM4G applied scenario construction to generate a set of alternative futures leading to 2030, each exercising different influences on the need for - and challenges associated with - development of SRM technologies. The scenarios then provided the context for the design of systems of governance with the capacity and legitimacy to respond to those challenges, and for the evaluation of the advantages and drawbacks of different options against a wide range of imaginary but plausible futures. SRM4G sought to initiate a conversation within the SRM research community on the capacity of foresight approaches to highlight the centrality of conceptions of the future to discussions of SRM's threats and opportunities, and in doing so, examined and challenged the assumptions embedded in conceptualizing SRM's aims, development and governance, and discussed the capacity of governance options to adapt to a wide range of possibilities.

12.
Proc Natl Acad Sci U S A ; 112(9): 2859-64, 2015 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-25691753

RESUMO

Following their synthesis in the endoplasmic reticulum (ER), voltage-gated sodium channels (NaV) are transported to the membranes of excitable cells, where they often cluster, such as at the axon initial segment of neurons. Although the mechanisms by which NaV channels form and maintain clusters have been extensively examined, the processes that govern their transport and degradation have received less attention. Our entry into the study of these processes began with the isolation of a new allele of the zebrafish mutant alligator, which we found to be caused by mutations in the gene encoding really interesting new gene (RING) finger protein 121 (RNF121), an E3-ubiquitin ligase present in the ER and cis-Golgi compartments. Here we demonstrate that RNF121 facilitates two opposing fates of NaV channels: (i) ubiquitin-mediated proteasome degradation and (ii) membrane localization when coexpressed with auxiliary NaVß subunits. Collectively, these results indicate that RNF121 participates in the quality control of NaV channels during their synthesis and subsequent transport to the membrane.


Assuntos
Proteólise , Domínios RING Finger , Ubiquitina-Proteína Ligases/metabolismo , Canais de Sódio Disparados por Voltagem/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/metabolismo , Animais , Sequência de Bases , Membrana Celular/genética , Membrana Celular/metabolismo , Dados de Sequência Molecular , Mutação , Complexo de Endopeptidases do Proteassoma/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Transporte Proteico/fisiologia , Ubiquitina/genética , Ubiquitina/metabolismo , Ubiquitina-Proteína Ligases/genética , Canais de Sódio Disparados por Voltagem/genética , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
13.
Philos Trans A Math Phys Eng Sci ; 372(2031)2014 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-25404678

RESUMO

We examine the claim that in governance for solar climate engineering research, and especially field tests, there is no need for external governance beyond existing mechanisms such as peer review and environmental impact assessments that aim to assess technically defined risks to the physical environment. By drawing on the historical debate on recombinant DNA research, we show that defining risks is not a technical question but a complex process of narrative formation. Governance emerges from within, and as a response to, narratives of what is at stake in a debate. In applying this finding to the case of climate engineering, we find that the emerging narrative differs starkly from the narrative that gave meaning to rDNA technology during its formative period, with important implications for governance. While the narrative of rDNA technology was closed down to narrowly focus on technical risks, that of climate engineering continues to open up and includes social, political and ethical issues. This suggests that, in order to be legitimate, governance must take into account this broad perception of what constitutes the relevant issues and risks of climate engineering, requiring governance that goes beyond existing mechanisms that focus on technical risks. Even small-scale field tests with negligible impacts on the physical environment warrant additional governance as they raise broader concerns that go beyond the immediate impacts of individual experiments.

14.
Dis Model Mech ; 5(6): 852-9, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22645112

RESUMO

Myotubular myopathy (MTM) is a severe congenital muscle disease characterized by profound weakness, early respiratory failure and premature lethality. MTM is defined by muscle biopsy findings that include centralized nuclei and disorganization of perinuclear organelles. No treatments currently exist for MTM. We hypothesized that aberrant neuromuscular junction (NMJ) transmission is an important and potentially treatable aspect of the disease pathogenesis. We tested this hypothesis in two murine models of MTM. In both models we uncovered evidence of a disorder of NMJ transmission: fatigable weakness, improved strength with neostigmine, and electrodecrement with repetitive nerve stimulation. Histopathological analysis revealed abnormalities in the organization, appearance and size of individual NMJs, abnormalities that correlated with changes in acetylcholine receptor gene expression and subcellular localization. We additionally determined the ability of pyridostigmine, an acetylcholinesterase inhibitor, to ameliorate aspects of the behavioral phenotype related to NMJ dysfunction. Pyridostigmine treatment resulted in significant improvement in fatigable weakness and treadmill endurance. In all, these results describe a newly identified pathological abnormality in MTM, and uncover a potential disease-modifying therapy for this devastating disorder.


Assuntos
Modelos Animais de Doenças , Miopatias Congênitas Estruturais/patologia , Miopatias Congênitas Estruturais/terapia , Junção Neuromuscular/patologia , Animais , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , Camundongos Knockout , Atividade Motora/efeitos dos fármacos , Miopatias Congênitas Estruturais/fisiopatologia , Neuregulina-1/metabolismo , Junção Neuromuscular/efeitos dos fármacos , Junção Neuromuscular/fisiopatologia , Junção Neuromuscular/ultraestrutura , Fenótipo , Brometo de Piridostigmina/farmacologia , Receptores Colinérgicos/genética , Receptores Colinérgicos/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Transmissão Sináptica/efeitos dos fármacos
15.
J Neurophysiol ; 108(1): 148-59, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22490555

RESUMO

The molecular and physiological basis of the touch-unresponsive zebrafish mutant fakir has remained elusive. Here we report that the fakir phenotype is caused by a missense mutation in the gene encoding voltage-gated calcium channel 2.1b (CACNA1Ab). Injection of RNA encoding wild-type CaV2.1 restores touch responsiveness in fakir mutants, whereas knockdown of CACNA1Ab via morpholino oligonucleotides recapitulates the fakir mutant phenotype. Fakir mutants display normal current-evoked synaptic communication at the neuromuscular junction but have attenuated touch-evoked activation of motor neurons. NMDA-evoked fictive swimming is not affected by the loss of CaV2.1b, suggesting that this channel is not required for motor pattern generation. These results, coupled with the expression of CACNA1Ab by sensory neurons, suggest that CaV2.1b channel activity is necessary for touch-evoked activation of the locomotor network in zebrafish.


Assuntos
Canais de Cálcio Tipo N/metabolismo , Ativação do Canal Iônico/genética , Tato/genética , Acetilcolina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/genética , Vias Aferentes/fisiologia , Animais , Animais Geneticamente Modificados , Bungarotoxinas/metabolismo , Canais de Cálcio Tipo N/genética , Curare/farmacologia , Relação Dose-Resposta a Droga , Embrião não Mamífero , Reação de Fuga/efeitos dos fármacos , Reação de Fuga/fisiologia , Potenciais Evocados/genética , Células HEK293 , Humanos , Ativação do Canal Iônico/efeitos dos fármacos , Leucina/genética , Locomoção/efeitos dos fármacos , Locomoção/genética , Modelos Moleculares , Morfolinas/farmacologia , Atividade Motora/genética , Neurônios Motores/efeitos dos fármacos , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/fisiologia , Mutagênese Sítio-Dirigida/métodos , Mutação/genética , Mutação de Sentido Incorreto/genética , Rede Nervosa/fisiologia , Antagonistas Nicotínicos/farmacologia , Medula Espinal/citologia , Medula Espinal/fisiologia , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/genética , Tato/fisiologia , Valina/genética , Peixe-Zebra , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
16.
J Biol Chem ; 287(2): 1080-9, 2012 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-22075003

RESUMO

In many tissues and organs, connexin proteins assemble between neighboring cells to form gap junctions. These gap junctions facilitate direct intercellular communication between adjoining cells, allowing for the transmission of both chemical and electrical signals. In rodents, gap junctions are found in differentiating myoblasts and are important for myogenesis. Although gap junctions were once believed to be absent from differentiated skeletal muscle in mammals, recent studies in teleosts revealed that differentiated muscle does express connexins and is electrically coupled, at least at the larval stage. These findings raised questions regarding the functional significance of gap junctions in differentiated muscle. Our analysis of gap junctions in muscle began with the isolation of a zebrafish motor mutant that displayed weak coiling at day 1 of development, a behavior known to be driven by slow-twitch muscle (slow muscle). We identified a missense mutation in the gene encoding Connexin 39.9. In situ hybridization found connexin 39.9 to be expressed by slow muscle. Paired muscle recordings uncovered that wild-type slow muscles are electrically coupled, whereas mutant slow muscles are not. The further examination of cellular activity revealed aberrant, arrhythmic touch-evoked Ca(2+) transients in mutant slow muscle and a reduction in the number of muscle fibers contracting in response to touch in mutants. These results indicate that Connexin 39.9 facilitates the spreading of neuronal inputs, which is irregular during motor development, beyond the muscle cells and that gap junctions play an essential role in the efficient recruitment of slow muscle fibers.


Assuntos
Conexinas/metabolismo , Proteínas Musculares/metabolismo , Mutação de Sentido Incorreto , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/metabolismo , Animais , Sequência de Bases , Conexinas/genética , Junções Comunicantes/genética , Junções Comunicantes/metabolismo , Regulação da Expressão Gênica/genética , Dados de Sequência Molecular , Fibras Musculares de Contração Lenta , Proteínas Musculares/genética , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
17.
J Neurosci ; 31(32): 11633-44, 2011 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-21832193

RESUMO

Mutations in the gene encoding TRPM7 (trpm7), a member of the Transient Receptor Potential (TRP) superfamily of cation channels that possesses an enzymatically active kinase at its C terminus, cause the touch-unresponsive zebrafish mutant touchdown. We identified and characterized a new allele of touchdown, as well as two previously reported alleles, and found that all three alleles harbor mutations that abolish channel activity. Through the selective restoration of TRPM7 expression in sensory neurons, we found that TRPM7's kinase activity and selectivity for divalent cations over monovalent cations were dispensable for touch-evoked activation of escape behaviors in zebrafish. Additional characterization revealed that sensory neurons were present and capable of responding to tactile stimuli in touchdown mutants, indicating that TRPM7 is not required for sensory neuron survival or mechanosensation. Finally, exposure to elevated concentrations of divalent cations was found to restore touch-evoked behaviors in touchdown mutants. Collectively, these findings are consistent with a role for zebrafish TRPM7 within sensory neurons in the modulation of neurotransmitter release at central synapses, similar to that proposed for mammalian TRPM7 at peripheral synapses.


Assuntos
Alelos , Reação de Fuga/fisiologia , Células Receptoras Sensoriais/fisiologia , Canais de Cátion TRPM/fisiologia , Tato/fisiologia , Proteínas de Peixe-Zebra/fisiologia , Animais , Animais Geneticamente Modificados , Feminino , Sistema Nervoso Periférico/citologia , Sistema Nervoso Periférico/metabolismo , Sistema Nervoso Periférico/fisiologia , Proteínas Serina-Treonina Quinases , Especificidade da Espécie , Canais de Cátion TRPM/genética , Tato/genética , Xenopus , Peixe-Zebra , Proteínas de Peixe-Zebra/genética
18.
J Neurosci ; 30(28): 9359-67, 2010 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-20631165

RESUMO

The process by which light touch in vertebrates is transformed into an electrical response in cutaneous mechanosensitive neurons is a largely unresolved question. To address this question we undertook a forward genetic screen in zebrafish (Danio rerio) to identify mutants exhibiting abnormal touch-evoked behaviors, despite the presence of sensory neurons and peripheral neurites. One family, subsequently named touché, was found to harbor a recessive mutation which produced offspring that were unresponsive to light touch, but responded to a variety of other sensory stimuli. The optogenetic activation of motor behaviors by touché mutant sensory neurons expressing channelrhodopsin-2 suggested that the synaptic output of sensory neurons was intact, consistent with a defect in sensory neuron activation. To explore sensory neuron activation we developed an in vivo preparation permitting the precise placement of a combined electrical and tactile stimulating probe upon eGFP-positive peripheral neurites. In wild-type larva electrical and tactile stimulation of peripheral neurites produced action potentials detectable within the cell body. In a subset of these sensory neurons an underlying generator potential could be observed in response to subthreshold tactile stimuli. A closer examination revealed that the amplitude of the generator potential was proportional to the stimulus amplitude. When assayed touché mutant sensory neurons also responded to electrical stimulation of peripheral neurites similar to wild-type larvae, however tactile stimulation of these neurites failed to uncover a subset of sensory neurons possessing generator potentials. These findings suggest that touché is required for generator potentials, and that cutaneous mechanoreceptors with generator potentials are necessary for responsiveness to light touch in zebrafish.


Assuntos
Potenciais Somatossensoriais Evocados/fisiologia , Células Receptoras Sensoriais/fisiologia , Transdução de Sinais/fisiologia , Tato/fisiologia , Proteínas de Peixe-Zebra/genética , Animais , Eletrofisiologia , Rede Nervosa/fisiologia , Neurônios Aferentes/fisiologia , Estimulação Física , Peixe-Zebra/genética
19.
Dev Neurobiol ; 70(7): 508-22, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20225246

RESUMO

A screen for zebrafish motor mutants identified two noncomplementing alleles of a recessive mutation that were named non-active (nav(mi89) and nav(mi130)). nav embryos displayed diminished spontaneous and touch-evoked escape behaviors during the first 3 days of development. Genetic mapping identified the gene encoding Na(V)1.6a (scn8aa) as a potential candidate for nav. Subsequent cloning of scn8aa from the two alleles of nav uncovered two missense mutations in Na(V)1.6a that eliminated channel activity when assayed heterologously. Furthermore, the injection of RNA encoding wild-type scn8aa rescued the nav mutant phenotype indicating that scn8aa was the causative gene of nav. In-vivo electrophysiological analysis of the touch-evoked escape circuit indicated that voltage-dependent inward current was decreased in mechanosensory neurons in mutants, but they were able to fire action potentials. Furthermore, tactile stimulation of mutants activated some neurons downstream of mechanosensory neurons but failed to activate the swim locomotor circuit in accord with the behavioral response of initial escape contractions but no swimming. Thus, mutant mechanosensory neurons appeared to respond to tactile stimulation but failed to initiate swimming. Interestingly fictive swimming could be initiated pharmacologically suggesting that a swim circuit was present in mutants. These results suggested that Na(V)1.6a was required for touch-induced activation of the swim locomotor network.


Assuntos
Sistema Nervoso Central/embriologia , Rede Nervosa/embriologia , Canais de Sódio/fisiologia , Natação/fisiologia , Tato/fisiologia , Proteínas de Peixe-Zebra/fisiologia , Peixe-Zebra/embriologia , Potenciais de Ação/genética , Potenciais de Ação/fisiologia , Animais , Comportamento Animal/fisiologia , Sistema Nervoso Central/crescimento & desenvolvimento , Sistema Nervoso Central/metabolismo , Mapeamento Cromossômico , Feminino , Masculino , Mecanorreceptores/citologia , Mecanorreceptores/metabolismo , Mutagênese Sítio-Dirigida , Mutação de Sentido Incorreto/genética , Canal de Sódio Disparado por Voltagem NAV1.6 , Rede Nervosa/crescimento & desenvolvimento , Rede Nervosa/metabolismo , Células Receptoras Sensoriais/citologia , Células Receptoras Sensoriais/metabolismo , Canais de Sódio/genética , Tato/genética , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/fisiologia , Proteínas de Peixe-Zebra/genética
20.
PLoS Genet ; 5(2): e1000372, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19197364

RESUMO

Myotubularin is a lipid phosphatase implicated in endosomal trafficking in vitro, but with an unknown function in vivo. Mutations in myotubularin cause myotubular myopathy, a devastating congenital myopathy with unclear pathogenesis and no current therapies. Myotubular myopathy was the first described of a growing list of conditions caused by mutations in proteins implicated in membrane trafficking. To advance the understanding of myotubularin function and disease pathogenesis, we have created a zebrafish model of myotubular myopathy using morpholino antisense technology. Zebrafish with reduced levels of myotubularin have significantly impaired motor function and obvious histopathologic changes in their muscle. These changes include abnormally shaped and positioned nuclei and myofiber hypotrophy. These findings are consistent with those observed in the human disease. We demonstrate for the first time that myotubularin functions to regulate PI3P levels in a vertebrate in vivo, and that homologous myotubularin-related proteins can functionally compensate for the loss of myotubularin. Finally, we identify abnormalities in the tubulo-reticular network in muscle from myotubularin zebrafish morphants and correlate these changes with abnormalities in T-tubule organization in biopsies from patients with myotubular myopathy. In all, we have generated a new model of myotubular myopathy and employed this model to uncover a novel function for myotubularin and a new pathomechanism for the human disease that may explain the weakness associated with the condition (defective excitation-contraction coupling). In addition, our findings of tubuloreticular abnormalities and defective excitation-contraction coupling mechanistically link myotubular myopathy with several other inherited muscle diseases, most notably those due to ryanodine receptor mutations. Based on our findings, we speculate that congenital myopathies, usually considered entities with similar clinical features but very disparate pathomechanisms, may at their root be disorders of calcium homeostasis.


Assuntos
Fibras Musculares Esqueléticas/ultraestrutura , Miopatias Congênitas Estruturais/etiologia , Miopatias Congênitas Estruturais/patologia , Proteínas Tirosina Fosfatases não Receptoras/fisiologia , Peixe-Zebra/genética , Animais , Modelos Animais de Doenças , Embrião não Mamífero/metabolismo , Imunofluorescência , Homeostase , Humanos , Fibras Musculares Esqueléticas/patologia , Músculo Esquelético/metabolismo , Mutação , Miopatias Congênitas Estruturais/metabolismo , Proteínas Tirosina Fosfatases não Receptoras/genética , Peixe-Zebra/metabolismo
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