250 research outputs found
Myosin Drives Retrograde F-Actin Flow in Neuronal Growth Cones
AbstractActin filaments assembled at the leading edge of neuronal growth cones are centripetally transported via retrograde F-actin flow, a process fundamental to growth cone guidance and other forms of directed cell motility. Here we investigated the role of myosins in retrograde flow, using two distinct modes of myosin inhibition: microinjection of NEM inactivated myosin S1 fragments, or treatment with 2,3-butanedione-2-monoxime, an inhibitor of myosin ATPase. Both treatments resulted in dose-dependent attenuation of retrograde F-actin flow and growth of filopodia. Growth was cytochalasin sensitive and directly proportional to the degree of myosin inhibition, suggesting that retrograde flow results from superimposition of two independent processes: actin assembly and myosin-based filament retraction. These results provide the first direct evidence for myosin involvement in neuronal growth cone function
A novel cytoskeletal structure involved in purse string wound closure and cell polarity maintenance.
The psychological science acceleratorβs COVID-19 rapid-response dataset
In response to the COVID-19 pandemic, the Psychological Science Accelerator coordinated three large-scale psychological studies to examine the effects of loss-gain framing, cognitive reappraisals, and autonomy framing manipulations on behavioral intentions and affective measures. The data collected (April to October 2020) included specific measures for each experimental study, a general questionnaire examining health prevention behaviors and COVID-19 experience, geographical and cultural context characterization, and demographic information for each participant. Each participant started the study with the same general questions and then was randomized to complete either one longer experiment or two shorter experiments. Data were provided by 73,223 participants with varying completion rates. Participants completed the survey from 111 geopolitical regions in 44 unique languages/dialects. The anonymized dataset described here is provided in both raw and processed formats to facilitate re-use and further analyses. The dataset offers secondary analytic opportunities to explore coping, framing, and self-determination across a diverse, global sample obtained at the onset of the COVID-19 pandemic, which can be merged with other time-sampled or geographic data.info:eu-repo/semantics/publishedVersio
Structural validity evidence for the Oxford Utilitarianism Scale across 15 languages
Background: The Psychological Science Accelerator (PSA) recently completed a large-scale moral psychology study using translated versions of the Oxford Utilitarianism Scale (OUS). However, the translated versions have no validity evidence. Objective: The study investigated the structural validity evidence of the OUS across 15 translated versions and produced version-specific validity reports. Methods: We analyzed OUS data from the PSA, which was collected internationally on a centralized online questionnaire. We also collected qualitative feedback from experts for each translated version. Results: For each version, we produced version-specific psychometric reports which include the following: (1) descriptive item and demographics analyses, (2) factor structure evidence using confirmatory factor analyses, (3) measurement invariance testing across languages using multiple-group confirmatory factor analyses and alignment optimization, and (4) reliability analyses using coefficients ? and ?.info:eu-repo/semantics/publishedVersio
The Role of Actin Turnover in Retrograde Actin Network Flow in Neuronal Growth Cones
The balance of actin filament polymerization and depolymerization maintains a steady state network treadmill in neuronal growth cones essential for motility and guidance. Here we have investigated the connection between depolymerization and treadmilling dynamics. We show that polymerization-competent barbed ends are concentrated at the leading edge and depolymerization is distributed throughout the peripheral domain. We found a high-to-low G-actin gradient between peripheral and central domains. Inhibiting turnover with jasplakinolide collapsed this gradient and lowered leading edge barbed end density. Ultrastructural analysis showed dramatic reduction of leading edge actin filament density and filament accumulation in central regions. Live cell imaging revealed that the leading edge retracted even as retrograde actin flow rate decreased exponentially. Inhibition of myosin II activity before jasplakinolide treatment lowered baseline retrograde flow rates and prevented leading edge retraction. Myosin II activity preferentially affected filopodial bundle disassembly distinct from the global effects of jasplakinolide on network turnover. We propose that growth cone retraction following turnover inhibition resulted from the persistence of myosin II contractility even as leading edge assembly rates decreased. The buildup of actin filaments in central regions combined with monomer depletion and reduced polymerization from barbed ends suggests a mechanism for the observed exponential decay in actin retrograde flow. Our results show that growth cone motility is critically dependent on continuous disassembly of the peripheral actin network
Alteration of the Cortical Actin Cytoskeleton Deregulates Ca2+ Signaling, Monospermic Fertilization, and Sperm Entry
Background: When preparing for fertilization, oocytes undergo meiotic maturation during which structural changes occur in the endoplasmic reticulum (ER) that lead to a more efficient calcium response. During meiotic maturation and subsequent fertilization, the actin cytoskeleton also undergoes dramatic restructuring. We have recently observed that rearrangements of the actin cytoskeleton induced by actin-depolymerizing agents, or by actin-binding proteins, strongly modulate intracellular calcium (Ca 2+) signals during the maturation process. However, the significance of the dynamic changes in F-actin within the fertilized egg has been largely unclear. Methodology/Principal Findings: We have measured changes in intracellular Ca 2+ signals and F-actin structures during fertilization. We also report the unexpected observation that the conventional antagonist of the InsP3 receptor, heparin, hyperpolymerizes the cortical actin cytoskeleton in postmeiotic eggs. Using heparin and other pharmacological agents that either hypo- or hyperpolymerize the cortical actin, we demonstrate that nearly all aspects of the fertilization process are profoundly affected by the dynamic restructuring of the egg cortical actin cytoskeleton. Conclusions/Significance: Our findings identify important roles for subplasmalemmal actin fibers in the process of spermegg interaction and in the subsequent events related to fertilization: the generation of Ca 2+ signals, sperm penetration
Causal circuit explanations of behavior: Are necessity and sufficiency necessary and sufficient?
In the current advent of technological innovation allowing for precise neural manipulations and copious data collection, it is hardly questioned that the explanation of behavioral processes is to be chiefly found in neural circuits. Such belief, rooted in the exhausted dualism of cause and effect, is enacted by a methodology that promotes βnecessity and sufficiencyβ claims as the goal-standard in neuroscience, thus instructing young students on what shall reckon as explanation. Here we wish to deconstruct and explicate the difference between what is done, what is said, and what is meant by such causal circuit explanations of behavior. Well-known to most philosophers, yet ignored or at least hardly ever made explicit by neuroscientists, the original grand claim of βunderstanding the brainβ is imperceptibly substituted by the methodologically sophisticated task of empirically establishing counterfactual dependencies. But for the 21st century neuroscientist, after so much pride, this is really an excess of humility. I argue that to upgrade intervention to explanation is prone to logical fallacies, interpretational leaps and carries a weak explanatory force, thus settling and maintaining low standards for intelligibility in neuroscience. To claim that behavior is explained by a βnecessary and sufficientβ neural circuit is, at best, misleading. In that, my critique (rather than criticism) is indeed mainly negative. Positively, I briefly suggest some available alternatives for conceptual progress, such as adopting circular causality (rather than lineal causality in the flavor of top-down reductionism), searching for principles of behavior(rather than taking an arbitrary definition of behavior and rushing to dissect its βunderlyingβ neural mechanisms), and embracing process philosophy (rather than substance-mechanistic ontologies). Overall, if the goal of neuroscience is to understand the relation between brain and behavior then, in addition to excruciating neural studies (one pillar), we will need a strong theory of behavior (the other pillar) and a solid foundation to establish their relation (the bridge)
An Order of Magnitude Faster AIP1-Associated Actin Disruption than Nucleation by the Arp2/3 Complex in Lamellipodia
The mechanism of lamellipod actin turnover is still under debate. To clarify the intracellular behavior of the recently-identified actin disruption mechanism, we examined kinetics of AIP1 using fluorescent single-molecule speckle microscopy. AIP1 is thought to cap cofilin-generated actin barbed ends. Here we demonstrate a reduction in actin-associated AIP1 in lamellipodia of cells overexpressing LIM-kinase. Moreover, actin-associated AIP1 was rapidly abolished by jasplakinolide, which concurrently blocked the F-actin-cofilin interaction. Jasplakinolide also slowed dissociation of AIP1, which is analogous to the effect of this drug on capping protein. These findings provide in vivo evidence of the association of AIP1 with barbed ends generated by cofilin-catalyzed filament disruption. Single-molecule observation found distribution of F-actin-associated AIP1 throughout lamellipodia, and revealed even faster dissociation of AIP1 than capping protein. The estimated overall AIP1-associated actin disruption rate, 1.8 Β΅M/s, was one order of magnitude faster than Arp2/3 complex-catalyzed actin nucleation in lamellipodia. This rate does not suffice the filament severing rate predicted in our previous high frequency filament severing-annealing hypothesis. Our data together with recent biochemical studies imply barbed end-preferred frequent filament disruption. Frequent generation of AIP1-associated barbed ends and subsequent release of AIP1 may be the mechanism that facilitates previously observed ubiquitous actin polymerization throughout lamellipodia
Effects of Ionomycin on Egg Activation and Early Development in Starfish
Ionomycin is a Ca2+-selective ionophore that is widely used to increase intracellular Ca2+ levels in cell biology laboratories. It is also occasionally used to activate eggs in the clinics practicing in vitro fertilization. However, neither the precise molecular action of ionomycin nor its secondary effects on the eggs' structure and function is well known. In this communication we have studied the effects of ionomycin on starfish oocytes and zygotes. By use of confocal microscopy, calcium imaging, as well as light and transmission electron microscopy, we have demonstrated that immature oocytes exposed to ionomycin instantly increase intracellular Ca2+ levels and undergo structural changes in the cortex. Surprisingly, when microinjected into the cells, ionomycin produced no Ca2+ increase. The ionomycin-induced Ca2+ rise was followed by fast alteration of the actin cytoskeleton displaying conspicuous depolymerization at the oocyte surface and in microvilli with concomitant polymerization in the cytoplasm. In addition, cortical granules were disrupted or fused with white vesicles few minutes after the addition of ionomycin. These structural changes prevented cortical maturation of the eggs despite the normal progression of nuclear envelope breakdown. At fertilization, the ionomycin-pretreated eggs displayed reduced Ca2+ response, no elevation of the fertilization envelope, and the lack of orderly centripetal translocation of actin fibers. These alterations led to difficulties in cell cleavage in the monospermic zygotes and eventually to a higher rate of abnormal development. In conclusion, ionomycin has various deleterious impacts on egg activation and the subsequent embryonic development in starfish. Although direct comparison is difficult to make between our findings and the use of the ionophore in the in vitro fertilization clinics, our results call for more defining investigations on the issue of a potential risk in artificial egg activation
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