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https://www.readbyqxmd.com/read/27321671/dynamic-axonal-translation-in-developing-and-mature-visual-circuits
#1
Toshiaki Shigeoka, Hosung Jung, Jane Jung, Benita Turner-Bridger, Jiyeon Ohk, Julie Qiaojin Lin, Paul S Amieux, Christine E Holt
Local mRNA translation mediates the adaptive responses of axons to extrinsic signals, but direct evidence that it occurs in mammalian CNS axons in vivo is scant. We developed an axon-TRAP-RiboTag approach in mouse that allows deep-sequencing analysis of ribosome-bound mRNAs in the retinal ganglion cell axons of the developing and adult retinotectal projection in vivo. The embryonic-to-postnatal axonal translatome comprises an evolving subset of enriched genes with axon-specific roles, suggesting distinct steps in axon wiring, such as elongation, pruning, and synaptogenesis...
June 30, 2016: Cell
https://www.readbyqxmd.com/read/26657954/ribotag-not-lost-in-translation
#2
REVIEW
Adam J Lesiak, John F Neumaier
No abstract text is available yet for this article.
January 2016: Neuropsychopharmacology: Official Publication of the American College of Neuropsychopharmacology
https://www.readbyqxmd.com/read/26054767/ribotag-is-a-flexible-tool-for-measuring-the-translational-state-of-targeted-cells-in-heterogeneous-cell-cultures
#3
Adam J Lesiak, Matthew Brodsky, John F Neumaier
Primary neuronal cultures are a useful tool for measuring pharmacological- and transgene-regulated gene expression; however, accurate measurements can be confounded by heterogeneous cell types and inconsistent transfection efficiency. Here we describe our adaptation of a ribosomal capture strategy that was designed to be used in transgenic mice expressing tagged ribosomal subunits (RiboTag) in specific cell types, thereby allowing measurement of translating RNAs from desired cell types within complex tissues...
June 2015: BioTechniques
https://www.readbyqxmd.com/read/25995477/opposing-role-for-egr3-in-nucleus-accumbens-cell-subtypes-in-cocaine-action
#4
Ramesh Chandra, T Chase Francis, Prasad Konkalmatt, Ariunzaya Amgalan, Amy M Gancarz, David M Dietz, Mary Kay Lobo
An imbalance in molecular signaling cascades and transcriptional regulation in nucleus accumbens (NAc) medium spiny neuron (MSN) subtypes, those enriched in dopamine D1 versus D2 receptors, is implicated in the behavioral responses to psychostimulants. To provide further insight into the molecular mechanisms occurring in MSN subtypes by cocaine, we examined the transcription factor early growth response 3 (Egr3). We evaluated Egr3 because it is a target of critical cocaine-mediated signaling pathways and because Egr3-binding sites are found on promoters of key cocaine-associated molecules...
May 20, 2015: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/25603796/cell-type-specific-gene-expression-profiling-in-brain-tissue-comparison-between-trap-lcm-and-rna-seq
#5
COMPARATIVE STUDY
TaeHyun Kim, Chae-Seok Lim, Bong-Kiun Kaang
The brain is an organ that consists of various cell types. As our knowledge of the structure and function of the brain progresses, cell type-specific research is gaining importance. Together with advances in sequencing technology and bioinformatics, cell type-specific transcriptome studies are providing important insights into brain cell function. In this review, we discuss 3 different cell type-specific transcriptome analyses i.e., Laser Capture Microdissection (LCM), Translating Ribosome Affinity Purification (TRAP)/RiboTag, and single cell RNA-Seq, that are widely used in the field of neuroscience...
July 2015: BMB Reports
https://www.readbyqxmd.com/read/25545461/drd2-cre-ribotag-mouse-line-unravels-the-possible-diversity-of-dopamine-d2-receptor-expressing-cells-of-the-dorsal-mouse-hippocampus
#6
Emma Puighermanal, Anne Biever, Julie Espallergues, Giuseppe Gangarossa, Dimitri De Bundel, Emmanuel Valjent
Increasing evidences suggest that dopamine facilitates the encoding of novel memories by the hippocampus. However, the role of dopamine D2 receptors (D2R) in such regulations remains elusive due to the lack of the precise identification of hippocampal D2R-expressing cells. To address this issue, mice expressing the ribosomal protein Rpl22 tagged with the hemagglutinin (HA) epitope were crossed with Drd2-Cre mice allowing the selective expression of HA in D2R-containing cells (Drd2-Cre:RiboTag mice). This new transgenic model revealed a more widespread pattern of D2R-expressing cells identified by HA immunoreactivity than the one initially reported in Drd2-EGFP mice, in which the hilar mossy cells were the main neuronal population detectable...
July 2015: Hippocampus
https://www.readbyqxmd.com/read/25486063/quantitative-profiling-of-initiating-ribosomes-in-vivo
#7
Xiangwei Gao, Ji Wan, Botao Liu, Ming Ma, Ben Shen, Shu-Bing Qian
Cells have evolved exquisite mechanisms to fine-tune the rate of protein synthesis in response to stress. Systemic mapping of start-codon positions and precise measurement of the corresponding initiation rate would transform our understanding of translational control. Here we present quantitative translation initiation sequencing (QTI-seq), with which the initiating ribosomes can be profiled in real time at single-nucleotide resolution. Resultant initiation maps not only delineated variations of start-codon selection but also highlighted a dynamic range of initiation rates in response to nutrient starvation...
February 2015: Nature Methods
https://www.readbyqxmd.com/read/25122893/ribosome-profiling-reveals-a-cell-type-specific-translational-landscape-in-brain-tumors
#8
Christian Gonzalez, Jennifer S Sims, Nicholas Hornstein, Angeliki Mela, Franklin Garcia, Liang Lei, David A Gass, Benjamin Amendolara, Jeffrey N Bruce, Peter Canoll, Peter A Sims
Glioma growth is driven by signaling that ultimately regulates protein synthesis. Gliomas are also complex at the cellular level and involve multiple cell types, including transformed and reactive cells in the brain tumor microenvironment. The distinct functions of the various cell types likely lead to different requirements and regulatory paradigms for protein synthesis. Proneural gliomas can arise from transformation of glial progenitors that are driven to proliferate via mitogenic signaling that affects translation...
August 13, 2014: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/24719406/murine-coagulation-factor-viii-is-synthesized-in-endothelial-cells
#9
Lesley A Everett, Audrey C A Cleuren, Rami N Khoriaty, David Ginsburg
The primary cellular source of factor VIII (FVIII) biosynthesis is controversial, with contradictory evidence supporting an endothelial or hepatocyte origin. LMAN1 is a cargo receptor in the early secretory pathway that is responsible for the efficient secretion of factor V (FV) and FVIII to the plasma. Lman1 mutations result in combined deficiency of FV and FVIII, with levels of both factors reduced to ~10% to 15% of normal in human patients. We generated Lman1 conditional knockout mice to characterize the FVIII secretion profiles of endothelial cells and hepatocytes...
June 12, 2014: Blood
https://www.readbyqxmd.com/read/24719255/riding-the-spermatogenic-wave-profiling-gene-expression-within-neonatal-germ-and-sertoli-cells-during-a-synchronized-initial-wave-of-spermatogenesis-in-mice
#10
Elizabeth Evans, Cathryn Hogarth, Debra Mitchell, Michael Griswold
Continual sperm production relies on germ cells undergoing spermatogenesis asynchronously. As a result, the testis always contains a mixed population of germ cells at different stages of their differentiation process. The heterogeneous nature of the testis makes profiling gene expression within Sertoli cells or specific populations of germ cells impossible when a wild-type testis is assessed. We recently reported a unique method for synchronizing spermatogenesis without affecting fertility by manipulating RA levels within the neonatal testis...
May 2014: Biology of Reproduction
https://www.readbyqxmd.com/read/24606126/genome-wide-identification-of-ar-regulated-genes-translated-in-sertoli-cells-in-vivo-using-the-ribotag-approach
#11
Karel De Gendt, Guido Verhoeven, Paul S Amieux, Miles F Wilkinson
An understanding of the molecular mechanisms by which androgens drive spermatogenesis has been thwarted by the fact that few consistent androgen receptor (AR) target genes have been identified. Here, we addressed this issue using next-generation sequencing coupled with the RiboTag approach, which purifies translated mRNAs expressed in cells that express cyclic recombinase (CRE). Using RiboTag mice expressing CRE in Sertoli cells (SCs), we identified genes expressed specifically in SCs in both prepubertal and adult mice...
April 2014: Molecular Endocrinology
https://www.readbyqxmd.com/read/23776628/ribotag-analysis-of-actively-translated-mrnas-in-sertoli-and-leydig-cells-in-vivo
#12
Elisenda Sanz, Ryan Evanoff, Albert Quintana, Elizabeth Evans, Jeremy A Miller, Chemyong Ko, Paul S Amieux, Michael D Griswold, G Stanley McKnight
Male spermatogenesis is a complex biological process that is regulated by hormonal signals from the hypothalamus (GnRH), the pituitary gonadotropins (LH and FSH) and the testis (androgens, inhibin). The two key somatic cell types of the testis, Leydig and Sertoli cells, respond to gonadotropins and androgens and regulate the development and maturation of fertilization competent spermatozoa. Although progress has been made in the identification of specific transcripts that are translated in Sertoli and Leydig cells and their response to hormones, efforts to expand these studies have been restricted by technical hurdles...
2013: PloS One
https://www.readbyqxmd.com/read/23505481/fmr1-transcript-isoforms-association-with-polyribosomes-regional-and-developmental-expression-in-mouse-brain
#13
David M Brackett, Feng Qing, Paul S Amieux, Drew L Sellers, Philip J Horner, David R Morris
The primary transcript of the mammalian Fragile X Mental Retardation-1 gene (Fmr1), like many transcripts in the central nervous system, is alternatively spliced to yield mRNAs encoding multiple proteins, which can possess quite different biochemical properties. Despite the fact that the relative levels of the 12 Fmr1 transcript isoforms examined here vary by as much as two orders of magnitude amongst themselves in both adult and embryonic mouse brain, all are associated with polyribosomes, consistent with translation into the corresponding isoforms of the protein product, FMRP (Fragile X Mental Retardation Protein)...
2013: PloS One
https://www.readbyqxmd.com/read/19666516/cell-type-specific-isolation-of-ribosome-associated-mrna-from-complex-tissues
#14
Elisenda Sanz, Linghai Yang, Thomas Su, David R Morris, G Stanley McKnight, Paul S Amieux
Gene profiling techniques allow the assay of transcripts from organs, tissues, and cells with an unprecedented level of coverage. However, most of these approaches are still limited by the fact that organs and tissues are composed of multiple cell types that are each unique in their patterns of gene expression. To identify the transcriptome from a single cell type in a complex tissue, investigators have relied upon physical methods to separate cell types or in situ hybridization and immunohistochemistry. Here, we describe a strategy to rapidly and efficiently isolate ribosome-associated mRNA transcripts from any cell type in vivo...
August 18, 2009: Proceedings of the National Academy of Sciences of the United States of America
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