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genetic engineering, neuron

Jung Eun Park, Xian Feng Zhang, Sang-Ho Choi, Junko Okahara, Erika Sasaki, Afonso C Silva
Chronic monitoring of neuronal activity in the living brain with optical imaging techniques became feasible owing to the continued development of genetically encoded calcium indicators (GECIs). Here we report for the first time the successful generation of transgenic marmosets (Callithrix jacchus), an important nonhuman primate model in neurophysiological research, which were engineered to express the green fluorescent protein (GFP)-based family of GECIs, GCaMP, under control of either the CMV or the hSyn promoter...
October 11, 2016: Scientific Reports
Susanna Alloisio, Patrizia Garbati, Federica Viti, Silvia Dante, Raffaella Barbieri, Giovanni Arnaldi, Alessia Petrelli, Arianna Gigoni, Paolo Giannoni, Rodolfo Quarto, Mario Nobile, Massimo Vassalli, Aldo Pagano
Recent advances in life sciences suggest that human and rodent cell responses to stimuli might differ significantly. In this context, the results achieved in neurotoxicology and biomedical research practices using neural networks obtained from mouse or rat primary culture of neurons would benefit of the parallel evaluation of the same parameters using fully differentiated neurons with a human genetic background, thus emphasizing the current need of neuronal cells with human origin. In this work, we developed a human functionally active neural network derived by human neuroblastoma cancer cells genetically engineered to overexpress NDM29, a non-coding RNA whose increased synthesis causes the differentiation toward a neuronal phenotype...
October 3, 2016: Molecular Neurobiology
Wei-Hsiang Huang, Casey J Guenthner, Jin Xu, Tiffany Nguyen, Lindsay A Schwarz, Alex W Wilkinson, Or Gozani, Howard Y Chang, Mehrdad Shamloo, Liqun Luo
Haploinsufficiency of Retinoic Acid Induced 1 (RAI1) causes Smith-Magenis syndrome (SMS), which is associated with diverse neurodevelopmental and behavioral symptoms as well as obesity. RAI1 encodes a nuclear protein but little is known about its molecular function or the cell types responsible for SMS symptoms. Using genetically engineered mice, we found that Rai1 preferentially occupies DNA regions near active promoters and promotes the expression of a group of genes involved in circuit assembly and neuronal communication...
September 22, 2016: Neuron
Helen H Yang, François St-Pierre
UNLABELLED: A longstanding goal in neuroscience is to understand how spatiotemporal patterns of neuronal electrical activity underlie brain function, from sensory representations to decision making. An emerging technology for monitoring electrical dynamics, voltage imaging using genetically encoded voltage indicators (GEVIs), couples the power of genetics with the advantages of light. Here, we review the properties that determine indicator performance and applicability, discussing both recent progress and technical limitations...
September 28, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Natalia V Barykina, Oksana M Subach, Danila A Doronin, Vladimir P Sotskov, Marina A Roshchina, Tatiana A Kunitsyna, Aleksey Y Malyshev, Ivan V Smirnov, Asya M Azieva, Ilya S Sokolov, Kiryl D Piatkevich, Mikhail S Burtsev, Anna M Varizhuk, Galina E Pozmogova, Konstantin V Anokhin, Fedor V Subach, Grigori N Enikolopov
Genetically encoded calcium indicators (GECIs) are mainly represented by two- or one-fluorophore-based sensors. One type of two-fluorophore-based sensor, carrying Opsanus troponin C (TnC) as the Ca(2+)-binding moiety, has two binding sites for calcium ions, providing a linear response to calcium ions. One-fluorophore-based sensors have four Ca(2+)-binding sites but are better suited for in vivo experiments. Herein, we describe a novel design for a one-fluorophore-based GECI with two Ca(2+)-binding sites. The engineered sensor, called NTnC, uses TnC as the Ca(2+)-binding moiety, inserted in the mNeonGreen fluorescent protein...
September 28, 2016: Scientific Reports
Chia-Yi Lin, Shih-Chuan Huang, Chun-Che Tung, Chih-Hsuan Chou, Susan Shur-Fen Gau, Hsien-Sung Huang
Genomic imprinting is an epigenetic mechanism causing monoallelic expression in a parent-of-origin-specific manner. Disruption of imprinted genes causes various neurological and psychiatric disorders. However, the role of imprinted genes in the brain is largely unknown. Different cell types within distinct brain regions can influence the genomic imprinting status, but imprinted genes in single cell types within distinct brain regions have not been characterized on a genome-wide scale. To address this critical question, we used a multi-stage approach, which combined genetically engineered mice with fluorescence-based laser capture microdissection (LCM) to capture excitatory neurons, inhibitory neurons and astrocytes as single cells in layer 2/3 of mouse visual cortex...
2016: PloS One
Ting-Hao Huang, Tarciso Velho, Carlos Lois
We used a synthetic genetic system based on ligand-induced intramembrane proteolysis to monitor cell-cell contacts in animals. Upon ligand-receptor interaction in sites of cell-cell contact, the transmembrane domain of an engineered receptor is cleaved by intramembrane proteolysis and releases a protein fragment that regulates transcription in the interacting partners. We demonstrate that the system can be used to regulate gene expression between interacting cells both in vitro and in vivo, in transgenic Drosophila We show that the system allows for detection of interactions between neurons and glia in the Drosophila nervous system...
September 22, 2016: Development
Wen-Chen Ji, Xiao-Wei Zhang, Yu-Sheng Qiu
Spinal cord injury usually leads to permanent disability, which could cause a huge financial problem to the patient. Up to now there is no effective method to treat this disease. The key of the treatment is to enable the damage zone axonal regeneration and luckily it could go through the damage zone; last a connection can be established with the target neurons. This study attempts to combine stem cell, material science and genetic modification technology together, by preparing two genes modified adipose-derived stem cells and inducing them into neuron direction; then by compositing them on the silk fibroin/chitosan scaffold and implanting them into the spinal cord injury model, seed cells can have features of neuron cells...
August 20, 2016: World Journal of Experimental Medicine
Ismael Valladolid-Acebes, Teresa Daraio, Kerstin Brismar, Tomas Hökfelt, Christina Bark
The exocytosis of signaling molecules from neuronal, neuroendocrine and endocrine cells is regulated by membrane fusion involving SNAP-25 and associated SNARE proteins. The importance of this process for metabolic control recently became evident by studies of mouse mutants genetically engineered to only express one of 2 closely related, alternatively-spliced variants of SNAP-25. The results showed that even minor differences in the function of proteins regulating exocytosis are sufficient to provoke metabolic disease, including hyperglycaemia, liver steatosis, adipocyte hypertrophy and obesity...
July 2016: Adipocyte
Robiul Islam, Angelo Keramidas, Li Xu, Nela Durisic, Pankaj Sah, Joseph W Lynch
The ability to control neuronal activation is rapidly advancing our understanding of brain function and is widely viewed as having eventual therapeutic application. Although several highly effective optogenetic, optochemical genetic, and chemogenetic techniques have been developed for this purpose, new approaches may provide better solutions for addressing particular questions and would increase the number of neuronal populations that can be controlled independently. An early chemogenetic neuronal silencing method employed a glutamate receptor Cl(-) channel engineered for activation by 1-3 nM ivermectin...
September 27, 2016: ACS Chemical Neuroscience
Johannes Steffen, Markus Krohn, Kristin Paarmann, Christina Schwitlick, Thomas Brüning, Rita Marreiros, Andreas Müller-Schiffmann, Carsten Korth, Katharina Braun, Jens Pahnke
Alzheimer's disease primarily occurs as sporadic disease and is accompanied with vast socio-economic problems. The mandatory basic research relies on robust and reliable disease models to overcome increasing incidence and emerging social challenges. Rodent models are most efficient, versatile, and predominantly used in research. However, only highly artificial and mostly genetically modified models are available. As these 'engineered' models reproduce only isolated features, researchers demand more suitable models of sporadic neurodegenerative diseases...
2016: Acta Neuropathologica Communications
Toshiharu Nagatsu, Ikuko Nagatsu
Tyrosine hydroxylase (TH), which was discovered at the National Institutes of Health (NIH) in 1964, is a tetrahydrobiopterin (BH4)-requiring monooxygenase that catalyzes the first and rate-limiting step in the biosynthesis of catecholamines (CAs), such as dopamine, noradrenaline, and adrenaline. Since deficiencies of dopamine and noradrenaline in the brain stem, caused by neurodegeneration of dopamine and noradrenaline neurons, are mainly related to non-motor and motor symptoms of Parkinson's disease (PD), we have studied human CA-synthesizing enzymes [TH; BH4-related enzymes, especially GTP-cyclohydrolase I (GCH1); aromatic L-amino acid decarboxylase (AADC); dopamine β-hydroxylase (DBH); and phenylethanolamine N-methyltransferase (PNMT)] and their genes in relation to PD in postmortem brains from PD patients, patients with CA-related genetic diseases, mice with genetically engineered CA neurons, and animal models of PD...
November 2016: Journal of Neural Transmission
Tom J H Ruigrok, Sven van Touw, Patrice Coulon
Apart from the genetically engineered, modified, strains of rabies virus (RABV), unmodified 'fixed' virus strains of RABV, such as the 'French' subtype of CVS11, are used to examine synaptically connected networks in the brain. This technique has been shown to have all the prerequisite characteristics for ideal tracing as it does not metabolically affect infected neurons within the time span of the experiment, it is transferred transneuronally in one direction only and to all types of neurons presynaptic to the infected neuron, number of transneuronal steps can be precisely controlled by survival time and it is easily detectable with a sensitive technique...
2016: Frontiers in Neural Circuits
Juan A Varela, Joana S Ferreira, Julien P Dupuis, Pauline Durand, Delphine Bouchet, Laurent Groc
Recent developments in single-molecule imaging have revealed many biological mechanisms, providing high spatial and temporal resolution maps of molecular events. In neurobiology, these techniques unveiled that plasma membrane neurotransmitter receptors and transporters laterally diffuse at the surface of cultured brain cells. The photostability of bright nanoprobes, such as quantum dots (QDs), has given access to neurotransmitter receptor tracking over long periods of time with a high spatial resolution. However, our knowledge has been restricted to cultured systems, i...
October 2016: Neurophotonics
Darren Carwardine, Liang-Fong Wong, James W Fawcett, Elizabeth M Muir, Nicolas Granger
A multitude of factors must be overcome following spinal cord injury (SCI) in order to achieve clinical improvement in patients. It is thought that by combining promising therapies these diverse factors could be combatted with the aim of producing an overall improvement in function. Chondroitin sulphate proteoglycans (CSPGs) present in the glial scar that forms following SCI present a significant block to axon regeneration. Digestion of CSPGs by chondroitinase ABC (ChABC) leads to axon regeneration, neuronal plasticity and functional improvement in preclinical models of SCI...
August 15, 2016: Journal of the Neurological Sciences
Chong-Chi Chiu, Yi-En Liao, Ling-Yu Yang, Jing-Ya Wang, David Tweedie, Hanuma K Karnati, Nigel H Greig, Jia-Yi Wang
Traumatic brain injury (TBI) is a leading cause of mortality and morbidity worldwide. Neuroinflammation is prominent in the short and long-term consequences of neuronal injuries that occur after TBI. Neuroinflammation involves the activation of glia, including microglia and astrocytes, to release inflammatory mediators within the brain, and the subsequent recruitment of peripheral immune cells. Various animal models of TBI have been developed that have proved valuable to elucidate the pathophysiology of the disorder and to assess the safety and efficacy of novel therapies prior to clinical trials...
July 2, 2016: Journal of Neuroscience Methods
Sarah K Denny, Dian Yang, Chen-Hua Chuang, Jennifer J Brady, Jing Shan Lim, Barbara M Grüner, Shin-Heng Chiou, Alicia N Schep, Jessika Baral, Cécile Hamard, Martine Antoine, Marie Wislez, Christina S Kong, Andrew J Connolly, Kwon-Sik Park, Julien Sage, William J Greenleaf, Monte M Winslow
Metastases are the main cause of cancer deaths, but the mechanisms underlying metastatic progression remain poorly understood. We isolated pure populations of cancer cells from primary tumors and metastases from a genetically engineered mouse model of human small cell lung cancer (SCLC) to investigate the mechanisms that drive the metastatic spread of this lethal cancer. Genome-wide characterization of chromatin accessibility revealed the opening of large numbers of distal regulatory elements across the genome during metastatic progression...
July 14, 2016: Cell
Alinda R Fernandes, Divya M Chari
Both neurotrophin-based therapy and neural stem cell (NSC)-based strategies have progressed to clinical trials for treatment of neurological diseases and injuries. Brain-derived neurotrophic factor (BDNF) in particular can confer neuroprotective and neuro-regenerative effects in preclinical studies, complementing the cell replacement benefits of NSCs. Therefore, combining both approaches by genetically-engineering NSCs to express BDNF is an attractive approach to achieve combinatorial therapy for complex neural injuries...
September 28, 2016: Journal of Controlled Release: Official Journal of the Controlled Release Society
Matthew Locke, Lindsay C Davies, Phil Stephens
: Progenitor cells derived from the oral mucosa lamina propria (OMLP-PCs) demonstrate an ability to differentiate into tissue lineages removed from their anatomical origin. This clonally derived population of neural-crest cells have demonstrated potential to differentiate along mesenchymal and neuronal cell lineages. OBJECTIVE: Significant efforts are being made to generate functioning muscle constructs for use in research and clinical tissue engineering. In this study we aimed to determine the myogenic properties of clonal populations of expanded OMLP-PCs...
June 11, 2016: Archives of Oral Biology
Alinda R Fernandes, Divya M Chari
Genetically engineered neural stem cell (NSC) transplant populations offer key benefits in regenerative neurology, for release of therapeutic biomolecules in ex vivo gene therapy. NSCs are 'hard-to-transfect' but amenable to 'magnetofection'. Despite the high clinical potential of this approach, the low and transient transfection associated with the large size of therapeutic DNA constructs is a critical barrier to translation. We demonstrate for the first time that DNA minicircles (small DNA vectors encoding essential gene expression components but devoid of a bacterial backbone, thereby reducing construct size versus conventional plasmids) deployed with magnetofection achieve the highest, safe non-viral DNA transfection levels (up to 54%) reported so far for primary NSCs...
September 28, 2016: Journal of Controlled Release: Official Journal of the Controlled Release Society
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