Activated AKT/PKB signaling in C. elegans uncouples temporally distinct outputs of DAF-2/insulin-like signaling

BACKGROUND: In the nematode, Caenorhabditis elegans, a conserved insulin-like signaling pathway controls larval development, stress resistance and adult lifespan. AGE-1, a homolog of the p110 catalytic subunit of phosphoinositide 3-kinases (PI3K) comprises the major known effector pathway downstream of the insulin receptor, DAF-2. Phospholipid products of AGE-1/PI3K activate AKT/PKB kinase signaling via PDK-1. AKT/PKB signaling antagonizes nuclear translocation of the DAF-16/FOXO transcription factor. Reduced AGE-1/PI3K signaling permits DAF-16 to direct dauer larval arrest and promote long lifespan in adult animals. In order to study the downstream effectors of AGE-1/PI3K signaling in C. elegans, we conducted a genetic screen for mutations that suppress the constitutive dauer arrest phenotype of age-1(mg109) animals. RESULTS: This report describes mutations recovered in a screen for suppressors of the constitutive dauer arrest (daf-C) phenotype of age-1(mg109). Two mutations corresponded to alleles of daf-16. Two mutations were gain-of-function alleles in the genes, akt-1 and pdk-1, encoding phosphoinositide-dependent serine/threonine kinases. A fifth mutation, mg227, located on chromosome X, did not correspond to any known dauer genes, suggesting that mg227 may represent a new component of the insulin pathway. Genetic epistasis analysis by RNAi showed that reproductive development in age-1(mg109);akt-1(mg247) animals was dependent on the presence of pdk-1. Similarly, reproductive development in age-1(mg109);pdk-1(mg261) animals was dependent on akt-1. However, reproductive development in age-1(mg109); mg227 animals required only akt-1, and pdk-1 activity was dispensable in this background. Interestingly, while mg227 suppressed dauer arrest in age-1(mg109) animals, it enhanced the long lifespan phenotype. In contrast, akt-1(mg247) and pdk-1(mg261) did not affect lifespan or stress resistance, while both daf-16 alleles fully suppressed these phenotypes. CONCLUSION: A screen for suppressors of PI3K mutant phenotypes identified activating mutations in two known pathway components, providing insights into their regulation. In particular, the interdependence of akt-1 and pdk-1, even in activated forms, supports the existence of AGE-1-independent pathways for these phospholipid-dependent kinases. Phenotypic analysis of these alleles shows that the larval and adult outputs of AGE-1/PI3K are fully separable in these mutants

Rendering an Account: An Open-State Archive in Postgraduate Supervision

The paper begins with a brief account of the transformation of research degree studies under the pressures of global capitalism and neo-liberal governmentality. A parallel transformation is occurring in the conduct of research through the use of information and communication technologies. Yet the potential of ICTs to shape practices of surveillance or to produce new student-supervisor relations and enhance the processes of developing the dissertation has received almost no critical attention. As doctoral supervisor and student, we then describe the features and uses of a web-based open state archive of the student's work-in-progress, developed by the student and accessible to his supervisor. Our intention was to encourage more open conversations between data and theorising, student and supervisor, and ultimately between the student and professional community. However, we recognise that relations of accountability, as these have developed within a contemporary "audit revolution" (Power, 1994, 1997) in universities, create particular "lines of visibility" (Munro, 1996). Thus while the open-state archive may help to redefine in less managerial terms notions of quality, transparency, flexibility and accountability, it might also make possible greater supervisory surveillance. How should we think about the panoptical potential of this archive? We argue that the diverse kinds of interactional patterns and pedagogical intervention it encourages help to create shifting subjectivities. Moreover, the archive itself is multiple, in bringing together an array of diverse materials that can be read in various ways, by following multiple paths. It therefore constitutes a collage, which we identify as a mode of cognition and of accounting distinct from but related to argument and narrative. As a more "open" text (Iser, 1978) it has an indeterminacy which may render it less open to abuse for the technologies of managerial accountability

Cities in fiction: Perambulations with John Berger

This paper explores selected novels by John Berger in which cities play a central role. These cities are places, partially real and partially imagined, where memory, hope, and despair intersect. My reading of the novels enables me to trace important themes in recent discourses on the nature of contemporary capitalism, including notions of resistance and universality. I also show how Berger?s work points to a writing that can break free from the curious capacity of capitalism to absorb and feed of its critique

‘Shared intelligibility’ and two reflexive strategies as methods of supporting ‘responsible decisions’ in a hermeneutic phenomenological study

Hermeneutic phenomenologists propose that researchers inescapably bring themselves into their research because interpretation must inevitably be influenced by their contexts and pre-understandings. They propose that interpretation is a dynamic and active part of the construction of a text’s meaning, and involvement in this construction process leads to deep empathic understanding of others’ experience, reappraisal of accepted social and cultural systems, and a level of self-enlightenment. The strengths of the hermeneutic methodological approach have led to its use in a number of disciplines, however, there remains concerns about interpretative validity. It is widely acknowledged that in order to support rigour and validity in hermeneutic studies, researchers are required to develop and integrate strategies within the research process to promote awareness of the interplay between their pre-understandings and interpretation. This paper discusses how episodic interviewing which capitalises on ‘shared intelligibility’, and the reflexive strategies of ‘oppositional arrangement of perspectives’ and ‘backgrounding’ were used to shed light on data from a study of the experiences and views of nursing home nurses regarding their occupational role and status, and work identity

The Inositol 5-Phosphatase dOCRL Controls PI(4,5)P2 Homeostasis and Is Necessary for Cytokinesis

SummaryDuring cytokinesis, constriction of an equatorial actomyosin ring physically separates the two daughter cells. At the cleavage furrow, the phosphoinositide PI(4,5)P2 plays an important role by recruiting and regulating essential proteins of the cytokinesis machinery [1]. Accordingly, perturbation of PI(4,5)P2 regulation leads to abortive furrowing and binucleation [2–4]. To determine how PI(4,5)P2 is regulated during cytokinesis, we individually knocked down each of the enzymes controlling the phosphoinositide (PIP) cycle in Drosophila. We show that depletion of the Drosophila ortholog of human oculocerebrorenal syndrome of Lowe 1 (OCRL1), an inositol 5-phosphatase mutated in the X-linked disorder oculocerebrorenal Lowe syndrome, triggers a high rate of cytokinesis failure. In absence of dOCRL, several essential components of the cleavage furrow were found to be incorrectly localized on giant cytoplasmic vacuoles rich in PI(4,5)P2 and in endocytic markers. We demonstrate that dOCRL is associated with endosomes and that it dephosphorylates PI(4,5)P2 on internal membranes to restrict this phosphoinositide at the plasma membrane and thereby regulates cleavage furrow formation and ingression. Identification of dOCRL as essential for cell division may be important to understand the molecular basis of the phenotypic manifestations of Lowe syndrome

Collaborative writing with young people with disabilities: raising new questions of authorship and agency

The process of communication between author and reader is a critical issue in examining any text. When considering the communication of ideas from young people whose voices are seldom heard, the journey from author to audience has particular significance. The construction of children and young people as ‘authors’ is important, especially for those with learning difficulties or who struggle to comply with the current emphasis on spelling, punctuation and grammar. This article relates to a UK Research-Council-funded three-year collaborative research project involving the co-creation of fictional stories with young people with disabilities to represent aspects of their lives. Drawing on frameworks from narratology, I analyse the co-creation of one of the stories, and present an interpretation and elaboration of the discourse structure of narrative fiction to illustrate the complexities of the relationship between the multi-faceted ‘author’ and community ‘reader’ of these stories. The combination of qualitative research and fictional prose has particular characteristics and implications for the dissemination and communication of research findings; while extending feminist critique of Barthes’ claim for the death of the author provides new insights for engaging children in writing with their own voice

Co-Regulation of the DAF-16 Target Gene, cyp-35B1/dod-13, by HSF-1 in C. elegans Dauer Larvae and daf-2 Insulin Pathway Mutants

Insulin/IGF-I-like signaling (IIS) has both cell autonomous and non-autonomous functions. In some cases, targets through which IIS regulates cell-autonomous functions, such as cell growth and metabolism, have been identified. In contrast, targets for many non-autonomous IIS functions, such as C. elegans dauer morphogenesis, remain elusive. Here, we report the use of genomic and genetic approaches to identify potential non-autonomous targets of C. elegans IIS. First, we used transcriptional microarrays to identify target genes regulated non-autonomously by IIS in the intestine or in neurons. C. elegans IIS controls expression of a number of stress response genes, which were differentially regulated by tissue-restricted IIS. In particular, expression of sod-3, a MnSOD enzyme, was not regulated by tissue-restricted IIS on the microarrays, while expression of hsp-16 genes was rescued back to wildtype by tissue restricted IIS. One IIS target regulated non-autonomously by age-1 was cyp-35B1/dod-13, encoding a cytochrome P450. Genetic analysis of the cyp-35B1 promoter showed both DAF-16 and HSF-1 are direct regulators. Based on these findings, we propose that hsf-1 may participate in the pathways mediating non-autonomous activities of age-1 in C. elegans

Narrative and Interpretation on Twitter: Reading tweets by telling stories

Existing research on communication on Twitter has largely ignored the question of how users make sense of the fragmentary tweets with which they are presented. Focusing on the use of Twitter for political reporting in post-revolutionary Egypt, this article argues that the production of mental stories provides readers with a mechanism for interpreting the meaning of individual tweets in terms of their relationships to other material. Drawing on contemporary narratology, it argues that Twitter exhibits key elements of narrativity, but that a creative reading process is nonetheless required to transform this incipient narrativity into coherent, sense-making mental narratives. This foregrounding of the reader’s creative role makes stories on Twitter highly fluid and dynamic. Through reference to classic critical theory, I propose that this nonetheless represents an evolution rather than a radical break from earlier forms of narrative reception, which in many cases demanded similarly creative reading practices