OLIG gene targeting in human pluripotent stem cells for motor neuron and oligodendrocyte differentiation.
AuthorsLiu Y, Jiang P, Deng W,
JournalNat Protoc
PubMed ID21527921
Pluripotent stem cells can be genetically labeled to facilitate differentiation studies. In this paper, we describe a gene-targeting protocol to knock in a GFP cassette into key gene loci in human pluripotent stem cells (hPSCs), and then use the genetically tagged hPSCs to guide in vitro differentiation, immunocytochemical and electrophysiological ... More
Panglial gap junctional communication is essential for maintenance of myelin in the CNS.
AuthorsTress O, Maglione M, May D, Pivneva T, Richter N, Seyfarth J, Binder S, Zlomuzica A, Seifert G, Theis M, Dere E, Kettenmann H, Willecke K,
JournalJ Neurosci
PubMed ID22649229
'In this study, we have investigated the contribution of oligodendrocytic connexin47 (Cx47) and astrocytic Cx30 to panglial gap junctional networks as well as myelin maintenance and function by deletion of both connexin coding DNAs in mice. Biocytin injections revealed complete disruption of oligodendrocyte-to-astrocyte coupling in the white matter of 10- ... More
Reaching out for signals: filopodia sense EGF and respond by directed retrograde transport of activated receptors.
AuthorsLidke DS, Lidke KA, Rieger B, Jovin TM, Arndt-Jovin DJ
JournalJ Cell Biol
PubMed ID16103229
'ErbB1 receptors situated on cellular filopodia undergo systematic retrograde transport after binding of the epidermal growth factor (EGF) and activation of the receptor tyrosine kinase. Specific inhibitors of the erbB1 receptor tyrosine kinase as well as cytochalasin D, a disruptor of the actin cytoskeleton, abolish transport but not free diffusion ... More
Layer-specific experience-dependent rewiring of thalamocortical circuits.
AuthorsWang L, Kloc M, Gu Y, Ge S, Maffei A,
JournalJ Neurosci
PubMed ID23447625
Thalamocortical circuits are central to sensory and cognitive processing. Recent work suggests that the thalamocortical inputs onto L4 and L6, the main input layers of neocortex, are activated differently by visual stimulation. Whether these differences depend on layer-specific organization of thalamocortical circuits; or on specific properties of synapses onto receiving ... More
Complex autonomous firing patterns of striatal low-threshold spike interneurons.
AuthorsBeatty JA, Sullivan MA, Morikawa H, Wilson CJ,
JournalJ Neurophysiol
PubMed ID22572945
During sensorimotor learning, tonically active neurons (TANs) in the striatum acquire bursts and pauses in their firing based on the salience of the stimulus. Striatal cholinergic interneurons display tonic intrinsic firing, even in the absence of synaptic input, that resembles TAN activity seen in vivo. However, whether there are other ... More
Functional identification of an outwardly rectifying pH- and anesthetic-sensitive leak K(+) conductance in hippocampal astrocytes.
Astrocytes function as spatial K(+) buffers by expressing a rich repertoire of K(+) channels. Earlier studies suggest that acid-sensitive tandem-pore K(+) channels, mainly TWIK-related acid-sensitive K(+) (TASK) channels, mediate part of the passive astroglial membrane conductance. Here, using a combination of electrophysiology and pharmacology, we investigated the presence of TASK-like ... More
Urotensin II modulates rapid eye movement sleep through activation of brainstem cholinergic neurons.
AuthorsHuitron-Resendiz S, Kristensen MP, Sánchez-Alavez M, Clark SD, Grupke SL, Tyler C, Suzuki C, Nothacker HP, Civelli O, Criado JR, Henriksen SJ, Leonard CS, de Lecea L,
JournalJ Neurosci
PubMed ID15944374
Urotensin II (UII) is a cyclic neuropeptide with strong vasoconstrictive activity in the peripheral vasculature. UII receptor mRNA is also expressed in the CNS, in particular in cholinergic neurons located in the mesopontine tegmental area, including the pedunculopontine tegmental (PPT) and lateral dorsal tegmental nuclei. This distribution suggests that the ... More
Urotensin II acts as a modulator of mesopontine cholinergic neurons.
AuthorsClark SD, Nothacker HP, Blaha CD, Tyler CJ, Duangdao DM, Grupke SL, Helton DR, Leonard CS, Civelli O,
JournalBrain Res
PubMed ID16183039
Urotensin II (UII) is a vasomodulatory peptide that was not predicted to elicit CNS activity. However, because we have recently shown that the urotensin II receptor (UII-R) is selectively expressed in rat mesopontine cholinergic (MPCh) neurons, we hypothesize that UII may have a central function. The present study demonstrates that ... More
Multiplexed protein analysis using encoded antibody-conjugated microbeads.
AuthorsTheilacker N, Roller EE, Barbee KD, Franzreb M, Huang X,
JournalJ R Soc Interface
PubMed ID21247950
We describe a method for multiplexed analysis of proteins using fluorescently encoded microbeads. The sensitivity of our method is comparable to the sensitivity obtained by enzyme-linked immunosorbent assay while only 5 µl sample volumes are needed. Streptavidin-coated, 1 µm beads are encoded with a combination of fluorophores at different intensity ... More
Evaluation of biotin-dye conjugates for use in an HPLC assay to assess relative binding of biotin derivatives with avidin and streptavidin.
In this investigation, studies were conducted to determine if size exclusion HPLC could be used to assess relative association rates (on-rates) and dissociation rates (off-rates) of biotin derivatives from avidin (Av) and streptavidin (SAv). For easy detection and quantification of biotin derivatives, molecules that can be detected by UV absorbance ... More
Blinking and nonradiant dark fraction of water-soluble quantum dots in aqueous solution.
Water-soluble quantum dots (qdots) are now being used in life sciences research to take advantage of their bright, easily excited fluorescence and high photostability. Although the frequent erratic blinking and substantial dark (never radiant) fractions that occur in all available qdots may interfere with many applications, these properties of individual ... More