Project description:Human NT2- cells differentiate in vitro into NT2N neurons in the presence of retinoic acid, and culture in the presence of mitosis inhibitors. We have previously observed the presence of the Lewisx determinant exclusively in NT2N neurons but not in NT2- cells. We also obtained evidence that Lex is synthesized in these cells by FUT9 (Brito,...,Costa (2007) J. Neurosci. Res. 85, 1260). We would like to compare the levels of expression of the different glycosyltransferases, particularly, fucosyltransferases and sialyltransferases to explain the exclusive detection of Lex in the neurons. In addition, we would like to know if there are some Lex-binding proteins that may act as binding partners of Lex in the neurons.
Project description:Neurons provide a rich setting for studying post-transcriptional control. Here, we investigate the landscape of translational control in neurons and search for mRNA features that explain differences in translational efficiency (TE), considering the interplay between TE, mRNA poly(A)-tail lengths, microRNAs, and neuronal activation. In neurons and brain tissues, TE correlates with tail length, and a few dozen mRNAs appear to undergo cytoplasmic polyadenylation upon light or chemical stimulation. However, the correlation between TE and tail length is modest, explaining <5% of TE variance, and even this modest relationship diminishes when accounting for other mRNA features. Thus, tail length appears to affect TE only minimally. Accordingly, miRNAs, which accelerate deadenylation of their mRNA targets, primarily influence target mRNA levels, with no detectable effect on either steady-state tail lengths or TE. Larger correlates with TE include codon composition and predicted mRNA folding energy. When combined in a model, the identified correlates explain 38–45% of TE variance. These results provide a framework for considering the relative impact of factors that contribute to translational control in neurons. They indicate that when examined in bulk, translational control in neurons largely resembles that of other types of post-embryonic cells. Thus, detection of more specialized control might require analyses that can distinguish translation occurring in neuronal processes from that occurring in cell bodies.
Project description:Exposure to electronic cigarette (e-cigarette) aerosol has been linked to a number of health concerns, including DNA damage, elevated oxidative stress, release of inflammatory cytokine, and dysfunctions in epithelial barriers. However, little is known about the effect of exclusive e-cigarette use on expression profiles of exosomal miRNAs, which play critical regulatory roles in many inflammatory responses and disease process including cancer. We aim to compare the exosomal microRNAs expression profile between exclusive e-cigarette users and normal controls without any tobacco product use (non-users). Using blood and urine samples from exclusive e-cigarette users and non-users in the Population Assessment of Tobacco and Health (PATH) Wave 1 study (2013-2014), we examined exosomal microRNAs expression levels through Illumina NextSeq 500/550 sequencing. We identified microRNAs that have significantly higher expression levels in exclusive e-cigarette users than non-users. Gene enrichment analysis of these significant exosomal microRNAs showed their involvement in cancer related pathways, which might indicate a potential elevated risk of cancer among exclusive e-cigarette users.
Project description:Microarray analysis has been applied to the study of ALS in order to investigate gene expression in whole spinal cord homogenates of SOD1 G93A mice and human ALS cases, although the massive presence of glial cells and inflammatory factors has made it difficult to define which gene expression changes were motor neuron specific. Recently, laser capture microdissection (LCM), combined with microarray analysis, has allowed the identification of motor neuron specific changes in gene expression in mouse and human ALS cases. The aim of the present study is to combine LCM and microarray analysis to compare the gene expression profiles of motor neurons from two SOD1G93A mouse strains (129Sv and C57) with different progression of the disease in order to discover the molecular mechanisms that may contribute to the distinct phenotypes and to uncover factors underlying fast and slow disease progression Motor neurons have been isolated from the spinal cord of 129SvG93A mice, C57G93A mice and non transgenic littermates at different time points and the transcription expression profile of the isolated motor neurons has been analysed
Project description:Cerebral cortex consists of many functional areas that are interconnected via direct axon projections from long association neurons. Long association neurons are located in layers 2/3, 5, and 6b. These neurons are intermingled with callosal neurons, which connect both cerebral hemisphere, in layers 2/3 and 5. To We used microarrays to compare gene expression profiles of long association neurons and callosal neurons in order to understand genetic programs that specifies these neuronal subtypes.
Project description:4 lymphoma cell lines (DOHH2, OCILy10, TMD8 and Toledo) have been treated with DMSO, CB103, NEO02734 or OTX015. The transcriptome has been sequenced after 6hrs from the treatment. The gene expression levels have been compared in order to elucidate the differencies and similarities for the different drugs compare to DMSO (control group).
Project description:We are interested in genes that determine the neuronal cellular fate and specific neurotransmitter phenotypes of cells in the nervous system. The reasons for our interest are two fold. First, identification of the genetic pathways operating in specific types of neurons could explain why they die in neurodegenerative diseases such as Alzheimer?s, Parkinson?s and amyotrophic lateral sclerosis. All of these disorders have in common the property that only certain types of neurotransmitter specific neurons degenerate. Secondly, neuronal cell replacement therapies using differentiated stem cell progeny hold the potential to reverse the devastating consequences of neurodegenerative diseases. The genetic personality of specific kinds of neurons must first be defined in order to use proper cells for neuronal replacement and this information will be essential in directing stem cell differentiation into proper developmental pathways. Our current approach to the problems of specification and neurotransmitter phenotype is to create transgenic animals where different neurotransmitter phenotypes are labeled with a fluorescent reporter gene. Labeled neurons are then isolated using Fluorescence Activated Cell Sorting. The purified populations of neurons are analyzed for their whole genome expression patterns using DNA microarray technology. We have successfully applied this approach using Drosophila cholinergic neurons and are now extending our observations to other classes of neurons that use GABA or glutamate as neurotransmitters. Cholinergic neurons express unique sets of ion channels, receptors and other types of genes. We also see unique sets of transcriptional regulatory proteins, and these may be important in the developmental pathways that result in the production of cholinergic neurons.
Project description:The hippocampus and entorhinal cortex are the earliest areas affected by AD pathology, which are related to early memory loss in patients. The hippocampus subfields are heterogeneous, with different order and degree affect by AD pathology. Studying the molecular changes may explain the difference, and may provide new therapeutic ideas for AD. In this study, specimens from the Chinese human brain banking were used to compare the proteome changes in the hippocampal subfields and the EC area affected by AD. The region-specific differential expressed proteins (associated with oligodendrocytes and myelin sheath) and co-variant proteins, also considered as constitute proteins (associated with astrocytes) were identified. The IHC results verified the specific loss of myelin in CA1, CA3 and EC areas, and find that protective S100A10-positive astrocytes number was increased in all hippocampal subfields and EC area. Further dual immunofluorescence results find that S100A10-positive astrocytes phagocyte apoptotic neuron debris. A hypothesis was put forward based on this, that S100A10-positive astrocytes protect neurons probably through engulfing apoptotic neurons. This study conducts a comprehensive proteomic analysis on the hippocampus subregion for the first time, highlighting the glia role in AD, and providing more information on the pathogenesis and therapy of AD.