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https://www.readbyqxmd.com/read/28423326/rapid-molecular-profiling-of-defined-cell-types-using-viral-trap
#1
Alexander R Nectow, Maria V Moya, Mats I Ekstrand, Awni Mousa, Kelly L McGuire, Caroline E Sferrazza, Bianca C Field, Gabrielle S Rabinowitz, Kirsty Sawicka, Yupu Liang, Jeffrey M Friedman, Nathaniel Heintz, Eric F Schmidt
Translational profiling methodologies enable the systematic characterization of cell types in complex tissues, such as the mammalian brain, where neuronal isolation is exceptionally difficult. Here, we report a versatile strategy for profiling CNS cell types in a spatiotemporally restricted fashion by engineering a Cre-dependent adeno-associated virus expressing an EGFP-tagged ribosomal protein (AAV-FLEX-EGFPL10a) to access translating mRNAs by translating ribosome affinity purification (TRAP). We demonstrate the utility of this AAV to target a variety of genetically and anatomically defined neural populations expressing Cre recombinase and illustrate the ability of this viral TRAP (vTRAP) approach to recapitulate the molecular profiles obtained by bacTRAP in corticothalamic neurons across multiple serotypes...
April 18, 2017: Cell Reports
https://www.readbyqxmd.com/read/28387242/chemical-labelling-for-visualizing-native-ampa-receptors-in-live-neurons
#2
Sho Wakayama, Shigeki Kiyonaka, Itaru Arai, Wataru Kakegawa, Shinji Matsuda, Keiji Ibata, Yuri L Nemoto, Akihiro Kusumi, Michisuke Yuzaki, Itaru Hamachi
The location and number of neurotransmitter receptors are dynamically regulated at postsynaptic sites. However, currently available methods for visualizing receptor trafficking require the introduction of genetically engineered receptors into neurons, which can disrupt the normal functioning and processing of the original receptor. Here we report a powerful method for visualizing native α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) which are essential for cognitive functions without any genetic manipulation...
April 7, 2017: Nature Communications
https://www.readbyqxmd.com/read/28386018/panin-neuroendocrine-cells-promote-tumorigenesis-via-neuronal-cross-talk
#3
Smrita Sinha, Ya-Yuan Fu, Adrien Grimont, Maren Ketcham, Kelly Lafaro, Joseph A Saglimbeni, Gokce Askan, Jennifer M Bailey, Jerry P Melchor, Yi Zhong, Min Geol Joo, Olivera Grbovic-Huezo, In-Hong Yang, Olca Basturk, Lindsey Baker, Young Park, Robert C Kurtz, David Tuveson, Steven D Leach, Pankaj J Pasricha
Nerves are a notable feature of the tumor microenvironment in some epithelial tumors, but their role in the malignant progression of pancreatic ductal adenocarcinoma (PDAC) is uncertain. Here, we identify dense innervation in the microenvironment of precancerous pancreatic lesions, known as pancreatic intraepithelial neoplasms (PanIN), and describe a unique subpopulation of neuroendocrine PanIN cells that express the neuropeptide substance P (SP) receptor neurokinin 1-R (NK1-R). Using organoid culture, we demonstrated that sensory neurons promoted the proliferation of PanIN organoids via SP-NK1-R signaling and STAT3 activation...
April 6, 2017: Cancer Research
https://www.readbyqxmd.com/read/28381446/cutaneous-trpm8-expressing-sensory-afferents-are-a-small-population-of-neurons-with-unique-firing-properties
#4
Michael P Jankowski, Kristofer K Rau, H Richard Koerber
It has been well documented that the transient receptor potential melastatin 8 (TRPM8) receptor is involved in environmental cold detection. The role that this receptor plays in nociception however, has been somewhat controversial since conflicting reports have shown different neurochemical identities and responsiveness of TRPM8 neurons. In order to functionally characterize cutaneous TRMP8 fibers, we used two ex vivo somatosensory recording preparations to functionally characterize TRPM8 neurons that innervate the hairy skin in mice genetically engineered to express GFP from the TRPM8 locus...
April 2017: Physiological Reports
https://www.readbyqxmd.com/read/28372274/stem-cell-roadmap%C3%A2-the-industrial-point-of%C3%A2-view
#5
Mazen Elzaabi, Agnès Thevenin, Pierre-Noël Lirsac
CELLforCURE is a French Contract Development and Manufacturing Organization (CDMO) dedicated to industrialization and process development for routine manufacturing, GMP manufacturing for clinical and commercial batches and regulatory services and associated logistics. CELLforCURE is a subsidiary of LFB Group.Stem cells fields of application gather cell and gene therapy as well as tissue engineering. According to VisionGain survey, cell therapy medicinal products will remain predominant in the future.Clinical trials are sponsored either by universities or private companies...
2017: Bio-medical Materials and Engineering
https://www.readbyqxmd.com/read/28368775/neuronal-erk-signaling-in-response-to-graphene-oxide-in-nematode-caenorhabditis-elegans
#6
Man Qu, Yunhui Li, Qiuli Wu, Yankai Xia, Dayong Wang
ERK signaling is one of the important mitogen activated protein kinases (MAPKs). However, the role of ERK signaling in the regulation of response to engineered nanomaterial exposure is still largely unclear. In this study, using in vivo assay system of Caenorhabditis elegans, we investigated the function of ERK signaling in response to graphene oxide (GO) exposure and the underlying molecular mechanism. GO exposure increased the expression of MEK-2/MEK and MPK-1/ERK in the ERK signaling pathway. Mutation of mek-2 or mpk-1 resulted in a susceptibility to GO toxicity...
April 3, 2017: Nanotoxicology
https://www.readbyqxmd.com/read/28366745/non-overlapping-neural-networks-in-hydra-vulgaris
#7
Christophe Dupre, Rafael Yuste
To understand the emergent properties of neural circuits, it would be ideal to record the activity of every neuron in a behaving animal and decode how it relates to behavior. We have achieved this with the cnidarian Hydra vulgaris, using calcium imaging of genetically engineered animals to measure the activity of essentially all of its neurons. Although the nervous system of Hydra is traditionally described as a simple nerve net, we surprisingly find instead a series of functional networks that are anatomically non-overlapping and are associated with specific behaviors...
April 24, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28366453/genetic-correction-of-sod1-mutant-ipscs-reveals-erk-and-jnk-activated-ap1-as-a-driver-of-neurodegeneration-in-amyotrophic-lateral-sclerosis
#8
Akshay Bhinge, Seema C Namboori, Xiaoyu Zhang, Antonius M J VanDongen, Lawrence W Stanton
Although mutations in several genes with diverse functions have been known to cause amyotrophic lateral sclerosis (ALS), it is unknown to what extent causal mutations impinge on common pathways that drive motor neuron (MN)-specific neurodegeneration. In this study, we combined induced pluripotent stem cells-based disease modeling with genome engineering and deep RNA sequencing to identify pathways dysregulated by mutant SOD1 in human MNs. Gene expression profiling and pathway analysis followed by pharmacological screening identified activated ERK and JNK signaling as key drivers of neurodegeneration in mutant SOD1 MNs...
April 11, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/28365983/engineering-pak1-allosteric-switches
#9
Onur Dagliyan, Andrei V Karginov, Sho Yagishita, Madeline E Gale, Hui Wang, Celine DerMardirossian, Claire M Wells, Nikolay V Dokholyan, Haruo Kasai, Klaus M Hahn
P21-activated kinases (PAKs) are important regulators of cell motility and morphology. It has been challenging to interrogate their functions because cells adapt to genetic manipulation of PAK, and because inhibitors act on multiple PAK isoforms. Here we describe genetically encoded PAK1 analogues that can be selectively activated by the membrane-permeable small molecule rapamycin. An engineered domain inserted away from the active site responds to rapamycin to allosterically control activity of the PAK1 isoform...
April 6, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28362378/combining-intravital-fluorescent-microscopy-ivfm-with-genetic-models-to-study-engraftment-dynamics-of-hematopoietic-cells-to-bone-marrow-niches
#10
Lin Wang, Malgorzata M Kamocka, Amy Zollman, Nadia Carlesso
Increasing evidence indicates that normal hematopoiesis is regulated by distinct microenvironmental cues in the BM, which include specialized cellular niches modulating critical hematopoietic stem cell (HSC) functions(1)(,)(2). Indeed, a more detailed picture of the hematopoietic microenvironment is now emerging, in which the endosteal and the endothelial niches form functional units for the regulation of normal HSC and their progeny(3)(,)(4)(,)(5). New studies have revealed the importance of perivascular cells, adipocytes and neuronal cells in maintaining and regulating HSC function(6)(,)(7)(,)(8)...
March 21, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28321439/overexpression-of-parkinson-s-disease-associated-mutation-lrrk2-g2019s-in-mouse-forebrain-induces-behavioral-deficits-and-%C3%AE-synuclein-pathology
#11
Yulan Xiong, Stewart Neifert, Senthilkumar S Karuppagounder, Jeannette N Stankowski, Byoung Dae Lee, Jonathan C Grima, Guanxing Chen, Han Seok Ko, Yunjong Lee, Debbie Swing, Lino Tessarollo, Ted M Dawson, Valina L Dawson
Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene have been identified as an unambiguous cause of late-onset, autosomal-dominant familial Parkinson's disease (PD) and LRRK2 mutations are the strongest genetic risk factor for sporadic PD known to date. A number of transgenic mice expressing wild-type or mutant LRRK2 have been described with varying degrees of LRRK2-related abnormalities and modest pathologies. None of these studies directly addressed the role of the kinase domain in the changes observed and none of the mice present with robust features of the human disease...
March 2017: ENeuro
https://www.readbyqxmd.com/read/28288335/how-to-control-cyclic-nucleotide-signaling-by-light
#12
REVIEW
Vera Jansen, Jan F Jikeli, Dagmar Wachten
Optogenetics allows to non-invasively manipulate cellular functions with spatio-temporal precision by combining genetic engineering with the control of protein function by light. Since the discovery of channelrhodopsin has pioneered the field, the optogenetic toolkit has been ever expanding and allows now not only to control neuronal activity by light, but rather a multitude of other cellular functions. One important application that has been established in recent years is the light-dependent control of second messenger signaling...
March 10, 2017: Current Opinion in Biotechnology
https://www.readbyqxmd.com/read/28280264/in-vitro-generation-of-mature-midbrain-type-dopamine-neurons-by-adjusting-exogenous-nurr1-and-foxa2-expressions-to-their-physiologic-patterns
#13
Taeho Kim, Jae-Jin Song, Lesly Puspita, Parvin Valiulahi, Jae-Won Shim, Sang-Hun Lee
Developmental information aids stem cell biologists in producing tissue-specific cells. Recapitulation of the developmental profile of a specific cell type in an in vitro stem cell system provides a strategy for manipulating cell-fate choice during the differentiation process. Nurr1 and Foxa2 are potential candidates for genetic engineering to generate midbrain-type dopamine (DA) neurons for experimental and therapeutic applications in Parkinson's disease (PD), as forced expression of these genes in neural stem/precursor cells (NPCs) yields cells with a complete battery of midbrain DA neuron-specific genes...
March 10, 2017: Experimental & Molecular Medicine
https://www.readbyqxmd.com/read/28238201/advances-in-optogenetic-and-chemogenetic-methods-to-study-brain-circuits-in-non-human-primates
#14
REVIEW
Adriana Galvan, Michael J Caiola, Daniel L Albaugh
Over the last 10 years, the use of opto- and chemogenetics to modulate neuronal activity in research applications has increased exponentially. Both techniques involve the genetic delivery of artificial proteins (opsins or engineered receptors) that are expressed on a selective population of neurons. The firing of these neurons can then be manipulated using light sources (for opsins) or by systemic administration of exogenous compounds (for chemogenetic receptors). Opto- and chemogenetic tools have enabled many important advances in basal ganglia research in rodent models, yet these techniques have faced a slow progress in non-human primate (NHP) research...
February 25, 2017: Journal of Neural Transmission
https://www.readbyqxmd.com/read/28198703/implantable-optoelectronic-probes-for-in-vivo-optogenetics
#15
Ege Iseri, Duygu Kuzum
More than a decade has passed since optics and genetics came together and lead to the emerging technologies of optogenetics. The advent of light-sensitive opsins made it possible to optically trigger the neurons into activation or inhibition by using visible light. The importance of spatiotemporally isolating a segment of a neural network and controlling nervous signaling in a precise manner has driven neuroscience researchers and engineers to invest great efforts in designing high precision in vivo implantable devices...
February 15, 2017: Journal of Neural Engineering
https://www.readbyqxmd.com/read/28191903/efficient-genome-editing-in-the-mouse-brain-by-local-delivery-of-engineered-cas9-ribonucleoprotein-complexes
#16
Brett T Staahl, Madhurima Benekareddy, Claire Coulon-Bainier, Ashwin A Banfal, Stephen N Floor, Jennifer K Sabo, Cole Urnes, Gabriela Acevedo Munares, Anirvan Ghosh, Jennifer A Doudna
We demonstrate editing of post-mitotic neurons in the adult mouse brain following injection of Cas9 ribonucleoprotein (RNP) complexes in the hippocampus, striatum and cortex. Engineered variants of Cas9 with multiple SV40 nuclear localization sequences enabled a tenfold increase in the efficiency of neuronal editing in vivo. These advances indicate the potential of genome editing in the brain to correct or inactivate the underlying genetic causes of neurological diseases.
February 13, 2017: Nature Biotechnology
https://www.readbyqxmd.com/read/28158225/visualizing-the-enteric-nervous-system-using-genetically-engineered-double-reporter-mice-comparison-with-immunofluorescence
#17
Yanfen Jiang, Hui Dong, Lars Eckmann, Elaine M Hanson, Katherine C Ihn, Ravinder K Mittal
BACKGROUND AND AIMS: The enteric nervous system (ENS) plays a crucial role in the control of gastrointestinal motility, secretion and absorption functions. Immunohistochemistry has been widely used to visualize neurons of the ENS for more than two decades. Genetically engineered mice that report specific proteins can also be used to visualize neurons of the ENS. The goal of our study was to develop a mouse that expresses fluorescent neuronal nitric oxide synthase (nNOS) and choline acetyltransferase (ChAT), the two proteins expressed in 95% of the ENS neurons...
2017: PloS One
https://www.readbyqxmd.com/read/28137877/camkii-modulates-sodium-current-in-neurons-from-epileptic-scn2a-mutant-mice
#18
Christopher H Thompson, Nicole A Hawkins, Jennifer A Kearney, Alfred L George
Monogenic epilepsies with wide-ranging clinical severity have been associated with mutations in voltage-gated sodium channel genes. In the Scn2a(Q54) mouse model of epilepsy, a focal epilepsy phenotype is caused by transgenic expression of an engineered NaV1.2 mutation displaying enhanced persistent sodium current. Seizure frequency and other phenotypic features in Scn2a(Q54) mice depend on genetic background. We investigated the neurophysiological and molecular correlates of strain-dependent epilepsy severity in this model...
February 14, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28133476/neuronal-activity-controls-the-development-of-interneurons-in-the-somatosensory-cortex
#19
Rachel Babij, Natalia De Marco Garcia
BACKGROUND: Neuronal activity in cortical areas regulates neurodevelopment by interacting with defined genetic programs to shape the mature central nervous system. Electrical activity is conveyed to sensory cortical areas via intracortical and thalamocortical neurons, and includes oscillatory patterns that have been measured across cortical regions. OBJECTIVE: In this work, we review the most recent findings about how electrical activity shapes the developmental assembly of functional circuitry in the somatosensory cortex, with an emphasis on interneuron maturation and integration...
December 2016: Frontiers in Biology
https://www.readbyqxmd.com/read/28132681/human-mesenchymal-stem-cells-genetically-engineered-to-overexpress-brain-derived-neurotrophic-factor-improve-outcomes-in-huntington-s-disease-mouse-models
#20
Kari Pollock, Heather Dahlenburg, Haley Nelson, Kyle D Fink, Whitney Cary, Kyle Hendrix, Geralyn Annett, Audrey Torrest, Peter Deng, Joshua Gutierrez, Catherine Nacey, Karen Pepper, Stefanos Kalomoiris, Johnathon D Anderson, Jeannine McGee, William Gruenloh, Brian Fury, Gerhard Bauer, Alexandria Duffy, Theresa Tempkin, Vicki Wheelock, Jan A Nolta
Huntington's disease (HD) is a fatal degenerative autosomal dominant neuropsychiatric disease that causes neuronal death and is characterized by progressive striatal and then widespread brain atrophy. Brain-derived neurotrophic factor (BDNF) is a lead candidate for the treatment of HD, as it has been shown to prevent cell death and to stimulate the growth and migration of new neurons in the brain in transgenic mouse models. BDNF levels are reduced in HD postmortem human brain. Previous studies have shown efficacy of mesenchymal stem/stromal cells (MSC)/BDNF using murine MSCs, and the present study used human MSCs to advance the therapeutic potential of the MSC/BDNF platform for clinical application...
May 2016: Molecular Therapy: the Journal of the American Society of Gene Therapy
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