Regulation of chemotropic guidance of nerve growth cones by microRNA.

BACKGROUND: The small non-coding microRNAs play an important role in development by regulating protein translation, but their involvement in axon guidance is unknown. Here, we investigated the role of microRNA-134 (miR-134) in chemotropic guidance of nerve growth cones. RESULTS: We found that miR-134 is highly expressed in the neural tube of Xenopus embryos. Fluorescent in situ hybridization also showed that miR-134 is enriched in the growth cones of Xenopus spinal neurons in culture. Importantly, overexpression of miR-134 mimics or antisense inhibitors blocked protein synthesis (PS)-dependent attractive responses of Xenopus growth cones to a gradient of brain-derived neurotrophic factor (BDNF). However, miR-134 mimics or inhibitors had no effect on PS-independent bidirectional responses of Xenopus growth cones to bone morphogenic protein 7 (BMP7). Our data further showed that Xenopus LIM kinase 1 (Xlimk1) mRNA is a potential target of miR-134 regulation. CONCLUSIONS: These findings demonstrate a role for miR-134 in translation-dependent guidance of nerve growth cones. Different guidance cues may act through distinct signaling pathways to elicit PS-dependent and -independent mechanisms to steer growth cones in response to a wide array of spatiotemporal cues during development.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Selective Role of the Catalytic PI3K Subunit p110β in Impaired Higher Order Cognition in Fragile X Syndrome

SummaryDistinct isoforms of the PI3K catalytic subunit have specialized functions in the brain, but their role in cognition is unknown. Here, we show that the catalytic subunit p110β plays an important role in prefrontal cortex (PFC)-dependent cognitive defects in mouse models of Fragile X syndrome (FXS), an inherited intellectual disability. FXS is caused by loss of function of the fragile X mental retardation protein (FMRP), which binds and translationally represses mRNAs. PFC-selective knockdown of p110β, an FMRP target that is translationally upregulated in FXS, reverses deficits in higher cognition in Fmr1 knockout mice. Genetic full-body reduction of p110β in Fmr1 knockout mice normalizes excessive PI3K activity, restores stimulus-induced protein synthesis, and corrects increased dendritic spine density and behavior. Notably, adult-onset PFC-selective Fmr1 knockdown mice show impaired cognition, which is rescued by simultaneous p110β knockdown. Our results suggest that FMRP-mediated control of p110β is crucial for neuronal protein synthesis and cognition

Fragile X Mental Retardation Protein is Involved in Protein Synthesis-Dependent Collapse of Growth Cones Induced by Semaphorin-3A

Fragile X syndrome, the most frequent form of familial mental retardation, is caused by mutation of the Fmr1 gene. Fmr1 encodes the fragile X mental retardation protein (FMRP), an mRNA binding protein regulating local, postsynaptic mRNA translation along dendrites necessary for long-term synaptic plasticity. However, recent studies on FMRP localization in axons and growth cones suggest a possible function in the regulation of local protein synthesis needed for axon guidance. Here, we have demonstrated that FMRP is involved in axonal and growth cone responses induced by the axon guidance factor, Semaphorin-3A (Sema3A). In cultured hippocampal neurons from wild type mice, Sema3A-induced growth cone collapse was protein synthesis-dependent. In contrast, Sema3A-induced growth cone collapse was attenuated in Fmr1 knock-out (KO) neurons and insensitive to protein synthesis inhibitors, suggesting that FMRP is involved in protein synthesis-dependent growth cone collapse. Sema3A increased phosphorylation of eukaryotic initiation factor 4E (eIF4E), an indicator of local translation, in distal axons and growth cones of wild type, but not Fmr1 KO neurons. Furthermore, Sema3A rapidly induced a protein synthesis-dependent increase in levels of microtubule associated protein 1B (MAP1B) in distal axons of wild type neurons, but this response was attenuated in Fmr1 KO neurons. These results suggest a possible role of FMRP to regulate local translation and axonal protein localization in response to Sema3A. This study reveals a new link between FMRP and semaphorin signaling in vitro, and raises the possibility that FMRP may have a critical role in semaphorin signaling in axon guidance during brain development

This Is The Story of a Man Named Stanley : Narratology, Authorship and Agency in The Stanley Parable

From Aristotle’s Rhetoric to Kenneth Burke’s more contemporary Grammar of Motives, classical rhetorical scholarship has traditionally focused on examining the strategies, styles, and best methods of persuasion through a focus on the individual rhetor, and how they can achieve their desired end. The rise of interactive technologies in the past twenty years have provided contemporary challenges to these classical theories. A player’s ability to interact with a game, translating into an ability to participate in the construction of a text, has now allowed the argument-building process to become co-creative...or so it would appear. This thesis examines the struggle for authorial control in the digital age by examining The Stanley Parable, a game that pits the player in direct opposition to the narrator. The symbolic struggle in The Stanley Parable not only questions the ways in which game stories are told, but the significance of the player interacting with a game’s story. It also raises questions about the significance of these choices and their role in constructing meaning. I argue that the introduction of such technologies creates new rhetorical opportunities that expose the importance of narrative structure, authorship and agency within persuasion. In doing so, I draw from the works of Michel Foucault, Roland Barthes, and Ian Bogost, to examine why “ownership” in storytelling is significant, and why The Stanley Parable serves as a significant, postmodern addition to the ways persuasion can occur

Compartmentalisation and localisation of the translation initiation factor (eIF) 4F complex in normally growing fibroblasts

Previous observations of association of mRNAs and ribosomes with subcellular structures highlight the importance of localised translation. However, little is known regarding associations between eukaryotic translation initiation factors and cellular structures within the cytoplasm of normally growing cells. We have used detergent-based cellular fractionation coupled with immunofluorescence microscopy to investigate the subcellular localisation in NIH3T3 fibroblasts of the initiation factors involved in recruitment of mRNA for translation, focussing on eIF4E, the mRNA cap-binding protein, the scaffold protein eIF4GI and poly(A) binding protein (PABP). We find that these proteins exist mainly in a soluble cytosolic pool, with only a subfraction tightly associated with cellular structures. However, this "associated" fraction was enriched in active "eIF4F" complexes (eIF4E.eIF4G.eIF4A.PABP). Immunofluorescence analysis reveals both a diffuse and a perinuclear distribution of eIF4G, with the perinuclear staining pattern similar to that of the endoplasmic reticulum. eIF4E also shows both a diffuse staining pattern and a tighter perinuclear stain, partly coincident with vimentin intermediate filaments. All three proteins localise to the lamellipodia of migrating cells in close proximity to ribosomes, microtubules, microfilaments and focal adhesions, with eIF4G and eIF4E at the periphery showing a similar staining pattern to the focal adhesion protein vinculin

MeCP2 Regulates the Synaptic Expression of a Dysbindin-BLOC-1 Network Component in Mouse Brain and Human Induced Pluripotent Stem Cell-Derived Neurons

Clinical, epidemiological, and genetic evidence suggest overlapping pathogenic mechanisms between autism spectrum disorder (ASD) and schizophrenia. We tested this hypothesis by asking if mutations in the ASD gene MECP2 which cause Rett syndrome affect the expression of genes encoding the schizophrenia risk factor dysbindin, a subunit of the biogenesis of lysosome-related organelles complex-1 (BLOC-1), and associated interacting proteins. We measured mRNA and protein levels of key components of a dysbindin interaction network by, quantitative real time PCR and quantitative immunohistochemistry in hippocampal samples of wild-type and Mecp2 mutant mice. In addition, we confirmed results by performing immunohistochemistry of normal human hippocampus and quantitative qRT-PCR of human inducible pluripotent stem cells (iPSCs)-derived human neurons from Rett syndrome patients. We defined the distribution of the BLOC-1 subunit pallidin in human and mouse hippocampus and contrasted this distribution with that of symptomatic Mecp2 mutant mice. Neurons from mutant mice and Rett syndrome patients displayed selectively reduced levels of pallidin transcript. Pallidin immunoreactivity decreased in the hippocampus of symptomatic Mecp2 mutant mice, a feature most prominent at asymmetric synapses as determined by immunoelectron microcopy. Pallidin immunoreactivity decreased concomitantly with reduced BDNF content in the hippocampus of Mecp2 mice. Similarly, BDNF content was reduced in the hippocampus of BLOC-1 deficient mice suggesting that genetic defects in BLOC-1 are upstream of the BDNF phenotype in Mecp2 deficient mice. Our results demonstrate that the ASD-related gene Mecp2 regulates the expression of components belonging to the dysbindin interactome and these molecular differences may contribute to synaptic phenotypes that characterize Mecp2 deficiencies and ASD.Fil: Larimore, Jennifer. Agnes Scott College; Estados UnidosFil: Ryder, Pearl V.. University of Emory; Estados UnidosFil: Kim, Kun Yong. University of Yale. School of Medicine; Estados UnidosFil: Ambrose, L. Alex. Agnes Scott College; Estados UnidosFil: Chapleau, Christopher. University Of Alabama; Estados UnidosFil: Calfa, Gaston Diego. University Of Alabama; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gross, Christina. University of Emory; Estados UnidosFil: Bassell, Gary J.. University of Emory; Estados UnidosFil: Pozzo Miller, Lucas. University Of Alabama; Estados UnidosFil: Smith, Yoland. University of Emory; Estados UnidosFil: Talbot, Konrad. The Pennsylvania State University; Estados UnidosFil: Park, In Hyun. University of Yale. School of Medicine; Estados UnidosFil: Faundez, Victor. University of Emory; Estados Unido

Marketing Leadership in a Knowledge Economy

Often the most valuable assets of a marketing driven firm are intangible assets such as a brand name, intellectual capital, and the expertise and knowledge of employees. The new breed of marketing leaders understand that it is important for employees to collaborate and be engaged and that leaders must be agents of change, creative, ethical, and global thinkers who can create learning organizations. The research reveals that organizations that are going to thrive in the knowledge economy are those that have marketing leaders who can build learning organizations, encourage diversity, and ensure employees are engaged in meaningful work

Storytelling And Reflective Pedagogy: Transforming Nursing Education Through Faculty Development

Nurse educators require pedagogical approaches beyond traditional methods to facilitate student learning of new competencies to practice in complex health care environments. However, little direction is available about how to effectively transform education. The purpose of this quality improvement project was to develop and implement steps to initiate change in both systems and processes of teaching and learning; to provide an efficient, sustainable method to incorporate transformative pedagogies through innovative faculty development; and, to collect outcomes of an e-Learning course to support teaching, using Kirkpatrick’s 4-level Model. An innovative course using storytelling and reflective pedagogy was developed to guide faculty into a transformative learning experience to challenge assumptions, gather insights, and raise questions about teaching practices. Pre- and post-course surveys captured data across three levels: satisfaction, knowledge and skill acquisition, and change in behavior. Forty-five participants were initially evaluated, while 31 were eligible for evaluation at three months. Follow-up survey results yielded a 42% response rate. Pre- and post-surveys were analyzed using a two-tailed, dependent t-test. Significant gains were recorded across all three areas (p<0.05), with large to medium effect size noted using Cohen’s d. Follow-up surveys revealed a significant change in knowledge (p<0.05), whereas the skill and attitude effect change were not statistically significant (p<0.05). Results suggest storytelling and reflective pedagogy are effective for faculty to confront and resolve actual and desired teaching practices, and that faculty placed value on reflection to facilitate self-awareness, question assumptions, and nurture ideas about personal and professional growth