Linking specification to differentiation:From proneural genes to the regulation of ciliogenesis

Much of developmental biology is concerned with the processes by which cells become committed to particular fates in a regulated fashion, whereas cell biology addresses, among other things, the variety of differentiated forms and functions that cells can acquire. One open question is how the regulators of the former process lead to attainment of the latter. “High-level” regulators of cell fate specification include the proneural factors, which drive cells to commit as precursors in the sensory nervous system. Recent research has concentrated on the gene expression events downstream of proneural factor function. Here we summarize this research and describe our own research that has provided clear links between a proneural factor, atonal and the cell biological program of ciliogenesis, which is a central aspect of sensory neuron differentiation

The function and regulation of the bHLH gene, cato, in Drosophila neurogenesis

Abstract Background bHLH transcription factors play many roles in neural development. cousin of atonal (cato) encodes one such factor that is expressed widely in the developing sensory nervous system of Drosophila. However, nothing definitive was known of its function owing to the lack of specific mutations. Results We characterised the expression pattern of cato in detail using newly raised antibodies and GFP reporter gene constructs. Expression is predominantly in sensory lineages that depend on the atonal and amos proneural genes. In lineages that depend on the scute proneural gene, cato is expressed later and seems to be particularly associated with the type II neurons. Consistent with this, we find evidence that cato is a direct target gene of Atonal and Amos, but not of Scute. We generated two specific mutations of cato. Mutant embryos show several defects in chordotonal sensory lineages, most notably the duplication of the sensory neuron, which appears to be caused by an extra cell division. In addition, we show that cato is required to form the single chordotonal organ that persists in atonal mutant embryos. Conclusions We conclude that although widely expressed in the developing PNS, cato is expressed and regulated very differently in different sensory lineages. Mutant phenotypes correlate with cato's major expression in the chordotonal sensory lineage. In these cells, we propose that it plays roles in sense organ precursor maintenance and/or identity, and in controlling the number of cell divisions in the neuronal branch of the lineage arising from these precursors.</p

A framework for initialising a dynamic clustering algorithm: ART2-A

Algorithms in the Adaptive Resonance Theory (ART) family adapt to structural changes in data as new information presents, making it an exciting candidate for dynamic online clustering of big health data. Its use however has largely been restricted to the signal processing field. In this paper we introduce an refinement of the ART2-A method within an adapted separation and concordance (SeCo) framework which has been shown to identify stable and reproducible solutions from repeated initialisations that also provides evidence for an appropriate number of initial clusters that best calibrates the algorithm with the data presented. The results show stable, reproducible solutions for a mix of real-world heath related datasets and well known benchmark datasets, selecting solutions which better represent the underlying structure of the data than using a single measure of separation. The scalability of the method and it's facility for dynamic online clustering makes it suitable for finding structure in big data

Multiple enhancers contribute to spatial but not temporal complexity in the expression of the proneural gene, amos

BACKGROUND: The regulation of proneural gene expression is an important aspect of neurogenesis. In the study of the Drosophila proneural genes, scute and atonal, several themes have emerged that contribute to our understanding of the mechanism of neurogenesis. First, spatial complexity in proneural expression results from regulation by arrays of enhancer elements. Secondly, regulation of proneural gene expression occurs in distinct temporal phases, which tend to be under the control of separate enhancers. Thirdly, the later phase of proneural expression often relies on positive autoregulation. The control of these phases and the transition between them appear to be central to the mechanism of neurogenesis. We present the first investigation of the regulation of the proneural gene, amos. RESULTS: Amos protein expression has a complex pattern and shows temporally distinct phases, in common with previously characterised proneural genes. GFP reporter gene constructs were used to demonstrate that amos has an array of enhancer elements up- and downstream of the gene, which are required for different locations of amos expression. However, unlike other proneural genes, there is no evidence for separable enhancers for the different temporal phases of amos expression. Using mutant analysis and site-directed mutagenesis of potential Amos binding sites, we find no evidence for positive autoregulation as an important part of amos control during neurogenesis. CONCLUSION: For amos, as for other proneural genes, a complex expression pattern results from the sum of a number of simpler sub-patterns driven by specific enhancers. There is, however, no apparent separation of enhancers for distinct temporal phases of expression, and this correlates with a lack of positive autoregulation. For scute and atonal, both these features are thought to be important in the mechanism of neurogenesis. Despite similarities in function and expression between the Drosophila proneural genes, amos is regulated in a fundamentally different way from scute and atonal

ENVIROSAT-2000 report: Federal agency satellite requirements

The requirement of Federal agencies, other than NOAA, for the data and services of civil operational environmental satellites (both polar orbiting and geostationary) are summarized. Agency plans for taking advantage of proposed future Earth sensing space systems, domestic and foreign, are cited also. Current data uses and future requirements are addressed as identified by each agency

Curriculum transformation with students as partners

8732142. That was my student ID as an undergraduate (now one of the authors). It was a number, not a name. It distinguished students from professors and all other teaching staff and, in a symbolic way, reminded us all of our firm place as students, as learners. There was a big power differential between students and teachers in the 1980s. What we learned was prescribed, transmitted and tested in implicit ways (no rubrics or marking criteria in those days) and rarely were our skills tested – just what we knew and could recall at a given time. Sometimes people say that teaching is an act. Indeed, sage on the stage suggests this precisely. But being a student is also an act. Students also assume roles and personas. If we want curriculum transformation, we seek to put a stop to acting – to engage students and staff in authentic learning. MIDAS is our curriculum transformation project in the Faculty of Engineering and Information Technology (FEIT) at UTS – More Innovative Design-Able Students. In MIDAS, we want students and teachers to be their authentic selves in a true teaching and learning partnership. MIDAS seeks mutual respect in people, not the fulfilment of roles. MIDAS doesn’t see students as numbers, but as partners, as people who can learn, contribute, inspire, teach and create … and it sees teachers as people who also learn, contribute, inspire, teach and creat

Use of q-values to Improve a Genetic Algorithm to Identify Robust Gene Signatures

Several approaches have been proposed for the analysis of DNA microarray datasets, focusing on the performance and robustness of the final feature subsets. The novelty of this paper arises in the use of q-values to pre-filter the features of a DNA microarray dataset identifying the most significant ones and including this information into a genetic algorithm for further feature selection. This method is applied to a lung cancer microarray dataset resulting in similar performance rates and greater robustness in terms of selected features (on average a 36.21% of robustness improvement) when compared to results of the standard algorithm

Exploring the cytotoxicity, uptake, cellular response, and proteomics of mono- and dinuclear DNA light-switch complexes

Drug resistance to platinum chemotherapeutics targeting DNA often involves abrogation of apoptosis, and has emerged as a significant challenge in modern, non-targeted chemotherapy. Consequently, there is great interest in the anti-cancer properties of metal complexes - particularly those that interact with DNA - and mechanisms of consequent cell death. Herein we compare a parent cytotoxic complex [Ru(phen)2(tpphz)]2+ [phen = 1,10-phenanthroline, tpphz = tetrapyridyl [3,2-a:2',3'-c:3'',2''-h:2''',3'''-j] phenazine], with a mononuclear analogue with modified intercalating ligand, [Ru(phen)2(taptp)]2+,[taptp = 4,5,9,18-tetraazaphenanthreno[9,10-b] triphenylene], and two structurally related di-nuclear, tpphz-bridged, heterometallic complexes, RuRe and RuPt. These changes result in a switch from intercalation to groove binding DNA interaction, concomitant reduction in cytotoxic potency, but no significant change in relative cytotoxicity toward platinum-resistant A2780CIS cancer cells, indicating that DNA interaction mode is not critical for the mechanism of platinum resistance. All variants exhibited a light-switch effect, which for the first time, was exploited to investigate timing of cell death by live cell microscopy. Surprisingly, cell death occurred rapidly as a consequence of oncosis, characterized by loss of cytoplasmic volume control, absence of significant mitochondrial membrane potential loss, and lack of activation of apoptotic cell death markers. Importantly, a novel, quantitative proteomic analysis of the A2780 cell genome following exposure to either mononuclear complex reveals changes in protein expression associated with global cell responses to oxidative stress, and DNA replication/repair cellular pathways. This combination of a multiple targeting modality and induction of a non-apoptotic death mechanism makes these complexes highly promising chemotherapeutic cytotoxicity leads

Forkhead Transcription Factor Fd3F Cooperates with Rfx to Regulate a Gene Expression Program for Mechanosensory Cilia Specialization

Cilia have evolved hugely diverse structures and functions to participate in a wide variety of developmental and physiological processes. Ciliary specialization requires differences in gene expression, but few transcription factors are known to regulate this, and their molecular function is unclear. Here, we show that the Drosophila Forkhead box (Fox) gene, fd3F, is required for specialization of the mechanosensory cilium of chordotonal (Ch) neurons. fd3F regulates genes for Ch-specific axonemal dyneins and TRPV ion channels, which are required for sensory transduction, and retrograde transport genes, which are required to differentiate their distinct motile and sensory ciliary zones. fd3F is reminiscent of vertebrate Foxj1, a motile cilia regulator, but fd3F regulates motility genes as part of a broader sensory regulation program. Fd3F cooperates with the pan-ciliary transcription factor, Rfx, to regulate its targets directly. This illuminates pathways involved in ciliary specialization and the molecular mechanism of transcription factors that regulate them