Potential of activated microglia as a source of dysregulated extracellular microRNAs contributing to neurodegeneration in amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is the most common form of motor neuron degeneration in adults, and several mechanisms underlying the disease pathology have been proposed. It has been shown that glia communicate with other cells by releasing extracellular vesicles containing proteins and nucleic acids, including microRNAs (miRNAs), which play a role in the post-transcriptional regulation of gene expression. Dysregulation of miRNAs is commonly observed in ALS patients, together with inflammation and an altered microglial phenotype. However, the role of miRNA-containing vesicles in microglia-to-neuron communication in the context of ALS has not been explored in depth. This review summarises the evidence for the presence of inflammation, pro-inflammatory microglia and dysregulated miRNAs in ALS, then explores how microglia may potentially be responsible for this miRNA dysregulation. The possibility of pro-inflammatory ALS microglia releasing miRNAs which may then enter neuronal cells to contribute to degeneration is also explored. Based on the literature reviewed here, microglia are a likely source of dysregulated miRNAs and potential mediators of neurodegenerative processes. Therefore, dysregulated miRNAs may be promising candidates for the development of therapeutic strategies

Fault-tolerant hardware designs and their reliability analysis

This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University.Fault-tolerance, which is a complement to fault prevention, is an effective method of achieving ultra-high reliability. By taking this approach fault free computation can be achieved despite the presence of fault in the system. In this thesis three new fault tolerant techniques are presented and their advantages over well known fault-tolerant strategies are shown. One of these new techniques achieves higher reliability than any other similar techniques presented in the literature. Generally fault-tolerant structures consist of four major blocks: the replicated modules, the disagreement and detection circuit, the switching circuit, and the voting mechanism. The most critical component in a fault-tolerant system is the voter because the final output of the system is computed by this component. This dissertation presents a new implementation for voters which reduces both the complexity and the occupied area on the chip. The structures of the three techniques developed in this work are such that the complexity of their switching mechanisms grows only linearly with the number of modules but the voting mechanism complexity increases significantly. This is a better approach than those schemes in which the switching complexity increases significantly and the voter's complexity remains constant or grows linearly with the number of modules because it is easier to implement a complex voter than a complex switch (voters have more regular structures). Extensive comparisons are made between different fault-tolerant techniques. A new reliability model is also developed for system reliability evaluation of the new designs. The results of these analyses are plotted, and the advantages of the new techniques are demonstrated. In the final part of the work an expert system is described which uses the knowledge acquired by these comparisons. This expert system is meant as a prototype of a component of a CAD tool which will act as an advisor on fault-tolerant techniques

Corrigendum: an overview of MicroRNAs as biomarkers of ALS

A Corrigendum on An Overview of MicroRNAs as Biomarkers of ALS by Joilin, G., Leigh, P. N., Newbury, S. F., and Hafezparast, M. (2019). Front. Neurol. 10:186. doi: 10.3389/fneur.2019.00186 In the original article, there was a mistake in Table 1 as published. Some of the miRNAs listed in the table were incorrectly placed in the wrong column and/or row. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated

A mathematical understanding of how cytoplasmic dynein walks on microtubules

Cytoplasmic dynein 1 (hereafter referred to simply as dynein) is a dimeric motor protein that walks and transports intracellular cargos towards the minus end of microtubules. In this article, we formulate, based on physical principles, a mechanical model to describe the stepping behaviour of cytoplasmic dynein walking on microtubules from the cell membrane towards the nucleus. Unlike previous studies on physical models of this nature, we base our formulation on the whole structure of dynein to include the temporal dynamics of the individual subunits such as the cargo ( for example, an endosome, vesicle or bead), two rings of six ATPase domains associated with diverse cellular activities (AAAþ rings) and the microtubule-binding domains which allow dynein to bind to microtubules. This mathematical framework allows us to examine experimental observations on dynein across a wide range of different species, as well as being able to make predictions on the temporal behaviour of the individual components of dynein not currently experimentally measured. Furthermore, we extend the model framework to include backward stepping, variable step size and dwelling. The power of our model is in its predictive nature; first it reflects recent experimental observations that dynein walks on microtubules using a weakly coordinated stepping pattern with predominantly not passing steps. Second, the model predicts that interhead coordination in the ATP cycle of cytoplasmic dynein is important in order to obtain the alternating stepping patterns and long run lengths seen in experiments

Neurobiology of axonal transport defects in motor neuron diseases: opportunities for translational research?

Intracellular trafficking of cargoes is an essential process to maintain the structure and function of all mammalian cell types, but especially of neurons because of their extreme axon/dendrite polarisation. Axonal transport mediates the movement of cargoes such as proteins, mRNA, lipids, membrane-bound vesicles and organelles that are mostly synthesised in the cell body and in doing so is responsible for their correct spatiotemporal distribution in the axon, for example at specialised sites such as nodes of Ranvier and synaptic terminals. In addition, axonal transport maintains the essential long-distance communication between the cell body and synaptic terminals that allows neurons to react to their surroundings via trafficking of for example signalling endosomes. Axonal transport defects are a common observation in a variety of neurodegenerative diseases, and mutations in components of the axonal transport machinery have unequivocally shown that impaired axonal transport can cause neurodegeneration (Reviewed in El-Kadi et al., 2007, De Vos et al., 2008; Millecamps and Julien, 2013). Here we review our current understanding of axonal transport defects and the role they play in motor neuron diseases (MNDs) with a specific focus on the most common form of MND, amyotrophic lateral sclerosis (ALS)

Automatic segmentation of focal adhesions from mouse embryonic fibroblasts

This work describes an automatic algorithm for the segmentation and quantification of focal adhesions from mouse embryonic fibroblasts. The main challenges solved by this algorithm are: the variability of the intensity of the focal adhesions, the detection of an outer ring, which distinguishes the cell periphery responsible for the cell migration, and the quantification of the characteristics of the focal adhesions. The algorithm detects maximal regions through gradients and uses a region-growing algorithm limited by intensity-based edges. The outer ring is calculated based on the average radial intensity from an extended centroid of the cell. Finally, traditional morphological characteristics are obtained to distinguish between two groups of cells. Two of the measurements employed showed statistical difference between two groups of cells

A Conceptual Rainfall-Runoff Model Using the Auto Calibrated NAM Models in the Sarisoo River

This paper describes the application of a conceptual rainfall runoff model to investigate the peak and monthly flows at the Sarisoo River Basin on the North West of Iran. The model was calibrated using measured stream flow data and then validated for three years. Calculations of level and time of peak flows are vital for designing structures downstream in the catchment areas. The simulated peak flows were occurring in the months of February in 2003, 2006 and 2007 with approximate values of 6.32, 9.35 and 6.13 m3s-1 respectively. After calibrating 9 NAM parameters using record data of daily rainfall, monthly evaporation and daily discharge in the period of 1th October 2003 to 31th March 2006 and validating the model daily discharges were calculated for 12 years. The outputs of the calibrated model are able to be used in the assessment of water resources management models like Mike Basin, WEAP… because they normally work based on monthly flows with a large time horizon. The results show that monthly averages of mean, maximum and minimum flows are about 10%, 2% and 33% less than daily computed Nash–Sutcliffe coefficients, all calculated over a period of 12 years. The optimum values of the 9 NAM parameters obtained during the calibration procedure are presented. The reliability of MIKE11 NAM was evaluated based on the Nash–Sutcliffe coefficient (R2), Root Mean Square Error (RMSE), peak flow (RMSE) and low flow (RMSE). The R2 obtained during this study is 0.74

Mouse cytoplasmic dynein intermediate chains: identification of new isoforms, alternative splicing and tissue distribution of transcripts

BACKGROUND: Intracellular transport of cargoes including organelles, vesicles, signalling molecules, protein complexes, and RNAs, is essential for normal function of eukaryotic cells. The cytoplasmic dynein complex is an important motor that moves cargos along microtubule tracks within the cell. In mammals this multiprotein complex includes dynein intermediate chains 1 and 2 which are encoded by two genes, Dync1i1 and Dync1i2. These proteins are involved in dynein cargo binding and dynein complexes with different intermediate chains bind to specific cargoes, although the mechanisms to achieve this are not known. The DYNC1I1 and DYNC1I2 proteins are translated from different splice isoforms, and specific forms of each protein are essential for the function of different dynein complexes in neurons. METHODOLOGY/PRINCIPAL FINDINGS: Here we have undertaken a systematic survey of the dynein intermediate chain splice isoforms in mouse, basing our study on mRNA expression patterns in a range of tissues, and on bioinformatics analysis of mouse, rat and human genomic and cDNA sequences. We found a complex pattern of alternative splicing of both dynein intermediate chain genes, with maximum complexity in the embryonic and adult nervous system. We have found novel transcripts, including some with orthologues in human and rat, and a new promoter and alternative non-coding exon 1 for Dync1i2. CONCLUSIONS/SIGNIFICANCE: These data, including the cloned isoforms will be essential for understanding the role of intermediate chains in the cytoplasmic dynein complex, particularly their role in cargo binding within individual tissues including different brain regions

Behavioral and other phenotypes in a cytoplasmic Dynein light intermediate chain 1 mutant mouse

The cytoplasmic dynein complex is fundamentally important to all eukaryotic cells for transporting a variety of essential cargoes along microtubules within the cell. This complex also plays more specialized roles in neurons. The complex consists of 11 types of protein that interact with each other and with external adaptors, regulators and cargoes. Despite the importance of the cytoplasmic dynein complex, we know comparatively little of the roles of each component protein, and in mammals few mutants exist that allow us to explore the effects of defects in dynein-controlled processes in the context of the whole organism. Here we have taken a genotype-driven approach in mouse (Mus musculus) to analyze the role of one subunit, the dynein light intermediate chain 1 (Dync1li1). We find that, surprisingly, an N235Y point mutation in this protein results in altered neuronal development, as shown from in vivo studies in the developing cortex, and analyses of electrophysiological function. Moreover, mutant mice display increased anxiety, thus linking dynein functions to a behavioral phenotype in mammals for the first time. These results demonstrate the important role that dynein-controlled processes play in the correct development and function of the mammalian nervous system

Expression of a pathogenic mutation of SOD1 sensitizes aprataxin-deficient cells and mice to oxidative stress and triggers hallmarks of premature ageing

Aprataxin (APTX) deficiency causes progressive cerebellar degeneration, ataxia and oculomotor apraxia in man. Cell free assays and crystal structure studies demonstrate a role for APTX in resolving 5'-adenylated nucleic acid breaks, however, APTX function in vertebrates remains unclear due to the lack of an appropriate model system. Here, we generated a murine model in which a pathogenic mutant of superoxide dismutase 1 (SOD1(G93A)) is expressed in an Aptx-/- mouse strain. We report a delayed population doubling and accelerated senescence in Aptx-/- primary mouse fibroblasts, which is not due to detectable telomere instability or cell cycle deregulation but is associated with a reduction in transcription recovery following oxidative stress. Expression of SOD1(G93A) uncovers a survival defect ex vivo in cultured cells and in vivo in tissues lacking Aptx. The surviving neurons feature numerous and deep nuclear envelope invaginations, a hallmark of cellular stress. Furthermore, they possess an elevated number of high-density nuclear regions and a concomitant increase in histone H3 K9 trimethylation, hallmarks of silenced chromatin. Finally, the accelerated cellular senescence was also observed at the organismal level as shown by down-regulation of insulin-like growth factor 1 (IGF-1), a hallmark of premature ageing. Together, this study demonstrates a protective role of Aptx in vivo and suggests that its loss results in progressive accumulation of DNA breaks in the nervous system, triggering hallmarks of premature ageing, systemically