Journal ArticleDOI
A single protocol to detect transcripts of various types and expression levels in neural tissue and cultured cells: in situ hybridization using digoxigenin-labelled cRNA probes.
TLDR
A simple non-radioactive in situ hybridization procedure for tissue sections and cultured cells using digoxigenin-labelled cRNA probes for the detection of various transcripts present at a wide range of expression levels in the central nervous system is developed.Abstract:
We have developed a simple non-radioactive in situ hybridization procedure for tissue sections and cultured cells using digoxigenin-labelled cRNA probes. This protocol can be applied for the detection of various transcripts present at a wide range of expression levels in the central nervous system. Cerebellar hybridization signals for transcripts estimated to be expressed at high (MBP, myelin basic protein), moderate (GluR1, subunit of AMPA/kainate sensitive glutamate receptors) and low (inositol polyphosphate-5-phosphatase) levels of abundance are demonstrated as examples. The sensitivity and cellular resolution were significantly improved by avoiding any ethanol treatment commonly used in other procedures. The localization of a labelled cell with respect to its environment is shown to be more easily assessed by counterstaining of the tissue with the nuclear dye Hoechst 33258. The present protocol can be combined with immunocytochemistry as demonstrated for glial fibrillary acidic protein (GFAP). All steps of the procedure, including preparation and labelling of the cRNA probes, pretreatment of tissue, hybridization and visualization of the labelled transcripts, are described in detail.read more
Citations
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An RNA-Sequencing Transcriptome and Splicing Database of Glia, Neurons, and Vascular Cells of the Cerebral Cortex
Ye Zhang,Kenian Chen,Steven A. Sloan,Mariko L. Bennett,Anja R. Scholze,Sean O'Keeffe,Hemali Phatnani,Paolo Guarnieri,Christine Caneda,Nadine Ruderisch,Shuyun Deng,Shane A. Liddelow,Chaolin Zhang,Richard Daneman,Tom Maniatis,Ben A. Barres,Jian Qian Wu +16 more
TL;DR: The authors' data provide clues as to how neurons and astrocytes differ in their ability to dynamically regulate glycolytic flux and lactate generation attributable to unique splicing of PKM2, the gene encoding the glycoleytic enzyme pyruvate kinase.
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Cloning and characterization of a mammalian proton-coupled metal-ion transporter
Hiromi Gunshin,Bryan Mackenzie,Urs V. Berger,Yoshimi Gunshin,Michael F. Romero,Walter F. Boron,Stephan Nussberger,John L. Gollan,Matthias A. Hediger +8 more
TL;DR: A new metal-ion transporter in the rat, DCT1, which has an unusually broad substrate range that includes Fe2+, Zn2+, Mn2+, Co2+, Cd2+, Cu2+, Ni2+ and Pb2+.
Journal ArticleDOI
Function of the chemokine receptor CXCR4 in haematopoiesis and in cerebellar development
TL;DR: This is the first demonstration of the involvement of a G-protein-coupled chemokine receptor in neuronal cell migration and patterning in the central nervous system and may be important for designing strategies to block HIV entry into cells and for understanding mechanisms of pathogenesis in AIDS dementia.
Journal ArticleDOI
The classical complement cascade mediates CNS synapse elimination.
Beth Stevens,Nicola J. Allen,Luis E. Vazquez,Gareth R. Howell,Karen S. Christopherson,Navid Nouri,Kristina D. Micheva,Adrienne K. Mehalow,Andrew D. Huberman,Benjamin K. Stafford,Alexander Sher,Alan Litke,John D. Lambris,Stephen J. Smith,Simon W. M. John,Ben A. Barres +15 more
TL;DR: It is shown that C1q, the initiating protein in the classical complement cascade, is expressed by postnatal neurons in response to immature astrocytes and is localized to synapses throughout the postnatal CNS and retina, supporting a model in which unwanted synapses are tagged by complement for elimination and suggesting that complement-mediated synapse elimination may become aberrantly reactivated in neurodegenerative disease.
Journal ArticleDOI
PPAR gamma is required for placental, cardiac, and adipose tissue development.
Yaacov Barak,Michael C. Nelson,Michael C. Nelson,Estelita S. Ong,Estelita S. Ong,Ying Jones,Pilar Ruiz-Lozano,Kenneth R. Chien,Alan Koder,Ronald M. Evans,Ronald M. Evans +10 more
TL;DR: Findings both confirm and expand the current known spectrum of physiological functions regulated by PPAR gamma, implicating a previously unrecognized dependence of the developing heart on a functional placenta.
References
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A glial progenitor cell that develops in vitro into an astrocyte or an oligodendrocyte depending on culture medium
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A family of AMPA-selective glutamate receptors
Kari Keinänen,William Wisden,Bernd Sommer,Pia Werner,Anne Herb,Todd A. Verdoorn,Bert Sakmann,Peter H. Seeburg +7 more
TL;DR: Four cloned cDNAs encoding 900-amino acid putative glutamate receptors with approximately 70 percent sequence identity were isolated from a rat brain cDNA library and in situ hybridization revealed differential expression patterns of the cognate mRNAs throughout the brain.
Journal ArticleDOI
Cloning by functional expression of a member of the glutamate receptor family.
TL;DR: A complementary DNA clone is isolated by screening a rat brain cDNA library for expression of kainate-gated ion channels in Xenopus oocytes which on expression in oocytes forms a functional ion channel possessing the electrophysiological and pharmacological properties of the kainates subtype of the glutamate receptor family in the mammalian central nervous system.
Journal ArticleDOI
Synthesis and incorporation of myelin polypeptides into CNS myelin.
TL;DR: It was found that crude myelin fractions are highly enriched in mRNAs coding for the MBPs but not in mRNA coding for PLP, which suggests that whereas the bound polysomes synthesizing PLP are largely confined to the cell body, free polysome synthesizing MBPs are concentrated in oligodendrocyte processes involved in myelination, which explains the immediate incorporation of MBPs into the developing myelin sheath.