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Journal of Visualized Experiments: JoVE

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https://www.readbyqxmd.com/read/28605393/drosophila-courtship-conditioning-as-a-measure-of-learning-and-memory
#1
Tom S Koemans, Cornelia Oppitz, Rogier A T Donders, Hans van Bokhoven, Annette Schenck, Krystyna Keleman, Jamie M Kramer
Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model organisms such as the fruit fly, Drosophila melanogaster. Drosophila is useful for understanding the basic neurobiology underlying cognitive deficits resulting from mutations in genes associated with human cognitive disorders, such as intellectual disability (ID) and autism. This work describes a methodology for testing learning and memory using a classic paradigm in Drosophila known as courtship conditioning...
June 5, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605386/simultaneous-transcranial-alternating-current-stimulation-and-functional-magnetic-resonance-imaging
#2
Kathleen A Williams, Yuranny Cabral-Calderin, Carsten Schmidt-Samoa, Christiane Anne Weinrich, Peter Dechent, Melanie Wilke
Transcranial alternating current stimulation (tACS) is a promising tool for noninvasive investigation of brain oscillations. TACS employs frequency-specific stimulation of the human brain through current applied to the scalp with surface electrodes. Most current knowledge of the technique is based on behavioral studies; thus, combining the method with brain imaging holds potential to better understand the mechanisms of tACS. Because of electrical and susceptibility artifacts, combining tACS with brain imaging can be challenging, however, one brain imaging technique that is well suited to be applied simultaneously with tACS is functional magnetic resonance imaging (fMRI)...
June 5, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605383/quadruple-immunostaining-of-the-olfactory-bulb-for-visualization-of-olfactory-sensory-axon-molecular-identity-codes
#3
Naoki Ihara, Yuji Ikegaya, Haruki Takeuchi
The mouse olfactory system is often used to study mechanisms of neural circuit formation because of its simple anatomical structure. An Olfactory Sensory Neuron (OSN) is a bipolar cell with a single dendrite and a single unbranched axon. An OSN expresses only one Olfactory Receptor (OR) gene, OSNs expressing a given type of OR converge their axons to a few sets of invariant glomeruli in the Olfactory Bulb (OB). A remarkable feature of OSN projection is that the expressed ORs play instructive roles in axonal projection...
June 5, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605373/modeling-fast-scan-cyclic-voltammetry-data-from-electrically-stimulated-dopamine-neurotransmission-data-using-qnsim1-0
#4
Rashed Harun, Christine M Grassi, Miranda J Munoz, Amy K Wagner
Central dopaminergic (DAergic) pathways have an important role in a wide range of functions, such as attention, motivation, and movement. Dopamine (DA) is implicated in diseases and disorders including attention deficit hyperactivity disorder, Parkinson's disease, and traumatic brain injury. Thus, DA neurotransmission and the methods to study it are of intense scientific interest. In vivo fast-scan cyclic voltammetry (FSCV) is a method that allows for selectively monitoring DA concentration changes with fine temporal and spatial resolution...
June 5, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605391/intranasal-delivery-of-therapeutic-stem-cells-to-glioblastoma-in-a-mouse-model
#5
Dou Yu, Gina Li, Maciej S Lesniak, Irina V Balyasnikova
The intrinsic tropism towards brain malignancies renders stem cells as promising carriers of therapeutic agents against malignant tumors. The delivery of therapeutic stem cells via the intranasal route is a recently discovered alternative strategy, with strong potential for clinical translation, due to its non-invasive nature compared to intracranial implantation or delivery via systemic routes. The lack of blood brain barrier further strengthens the therapeutic potential of stem cells undergoing intranasal brain entry...
June 4, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605372/a-component-resolved-diagnostic-approach-for-a-study-on-grass-pollen-allergens-in-chinese-southerners-with-allergic-rhinitis-and-or-asthma
#6
Wenting Luo, Guichang Pan, Huimin Huang, Peiyan Zheng, Nili Wei, Yifei Zhang, Guangqiao Zeng, Baoqing Sun
Sensitization to grass pollen imposes a global risk for allergic airway diseases. Although prevention relies on local investigation of the pollen allergens, data on this topic are limited in southern China. Any available data were obtained by self-report questionnaires, skin prick tests, and total or specific IgE tests using crude extracts. For many reasons, these methods are unreliable. Serum sIgE reactivity to Bermuda grass, Timothy grass, and Humulus scandens allergens in a cohort of patients from Greater Guangzhou (southern China's largest city and its outskirts) with allergic rhinitis and/or asthma were examined using a fully-automated immunoassay analyzer as a component-resolved diagnostics (CRD) tool...
June 4, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605368/murine-short-axis-ventricular-heart-slices-for-electrophysiological-studies
#7
Gabriel Peinkofer, Juergen Hescheler, Marcel Halbach
Murine cardiomyocytes have been extensively used for in vitro studies of cardiac physiology and new therapeutic strategies. However, multicellular preparations of dissociated cardiomyocytes are not representative of the complex in vivo structure of cardiomyocytes, non-myocytes and extracellular matrix, which influences both mechanical and electrophysiological properties of the heart. Here we describe a technique to prepare viable ventricular slices of adult mouse hearts with a preserved in vivo like tissue structure, and demonstrate their suitability for electrophysiological recordings...
June 4, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605392/transfer-of-mammary-gland-forming-ability-between-mammary-basal-epithelial-cells-and-mammary-luminal-cells-via-extracellular-vesicles-exosomes
#8
Meng-Chieh Lin, Shih-Yin Chen, Pei-Lin He, Wen-Ting Luo, Hua-Jung Li
Cells can communicate via exosomes, ~100-nm extracellular vesicles (EVs) that contain proteins, lipids, and nucleic acids. Non-adherent/mesenchymal mammary epithelial cell (NAMEC)-derived extracellular vesicles can be isolated from NAMEC medium via differential ultracentrifugation. Based on their density, EVs can be purified via ultracentrifugation at 110,000 x g. The EV preparation from ultracentrifugation can be further separated using a continuous density gradient to prevent contamination with soluble proteins...
June 3, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605389/genome-wide-mapping-of-protein-dna-interactions-with-chec-seq-in-saccharomyces-cerevisiae
#9
Sebastian Grünberg, Gabriel E Zentner
Genome-wide mapping of protein-DNA interactions is critical for understanding gene regulation, chromatin remodeling, and other chromatin-resident processes. Formaldehyde crosslinking followed by chromatin immunoprecipitation and high-throughput sequencing (X-ChIP-seq) has been used to gain many valuable insights into genome biology. However, X-ChIP-seq has notable limitations linked to crosslinking and sonication. Native ChIP avoids these drawbacks by omitting crosslinking, but often results in poor recovery of chromatin-bound proteins...
June 3, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605371/precise-cellular-ablation-approach-for-modeling-acute-kidney-injury-in-developing-zebrafish
#10
Rohan Datta, Ada Wong, Troy Camarata, Farhana Tamanna, Imran Ilahi, Aleksandr Vasilyev
Acute Kidney Injury (AKI) is a common medical condition with a high mortality rate. With the repair abilities of the kidney, it is possible to restore adequate kidney function after supportive treatment. However, a better understanding of how nephron cell death and repair occur on the cellular level is required to minimize cell death and to enhance the regenerative process. The zebrafish pronephros is a good model system to accomplish this goal because it contains anatomical segments that are similar to the mammalian nephron...
June 3, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605394/a-novel-method-for-in-situ-electromechanical-characterization-of-nanoscale-specimens
#11
Russell C Reid, Alberto Piqué, Wonmo Kang
Electrically assisted deformation (EAD) is increasingly being used to improve the formability of metals during processes such as sheet metal rolling and forging. Adoption of this technique is proceeding despite disagreement concerning the underlying mechanism responsible for EAD. The experimental procedure described herein enables a more explicit study compared to previous EAD research by removing thermal effects, which are responsible for disagreement in interpreting previous EAD results. Furthermore, as the procedure described here enables EAD observation in situ and in real time in a transmission electron microscope (TEM), it is superior to existing post-mortem methods that observe EAD effects post-test...
June 2, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605388/2d-and-3d-matrices-to-study-linear-invadosome-formation-and-activity
#12
Julie Di Martino, Elodie Henriet, Zakaria Ezzoukhry, Chandrani Mondal, Jose Javier Bravo-Cordero, Violaine Moreau, Frederic Saltel
Cell adhesion, migration, and invasion are involved in many physiological and pathological processes. For example, during metastasis formation, tumor cells have to cross anatomical barriers to invade and migrate through the surrounding tissue in order to reach blood or lymphatic vessels. This requires the interaction between cells and the extracellular matrix (ECM). At the cellular level, many cells, including the majority of cancer cells, are able to form invadosomes, which are F-actin-based structures capable of degrading ECM...
June 2, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605367/fabrication-of-polymer-microspheres-for-optical-resonator-and-laser-applications
#13
Yohei Yamamoto, Daichi Okada, Soh Kushida, Zakarias Seba Ngara, Osamu Oki
This paper describes three methods of preparing fluorescent microspheres comprising π-conjugated or non-conjugated polymers: vapor diffusion, interface precipitation, and mini-emulsion. In all methods, well-defined, micrometer-sized spheres are obtained from a self-assembling process in solution. The vapor diffusion method can result in spheres with the highest sphericity and surface smoothness, yet the types of the polymers able to form these spheres are limited. On the other hand, in the mini-emulsion method, microspheres can be made from various types of polymers, even from highly crystalline polymers with coplanar, π-conjugated backbones...
June 2, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605359/production-of-metal-nanoparticles-by-pulsed-laser-ablation-in-liquids-a-tool-for-studying-the-antibacterial-properties-of-nanoparticles
#14
Matthew Ratti, Joseph J Naddeo, Julianne C Griepenburg, Sean M O'Malley, Daniel M Bubb, Eric A Klein
The emergence of multidrug-resistant bacteria is a global clinical concern leading some to speculate about our return to a "pre-antibiotics" era of medicine. In addition to efforts to identify novel small-molecule antimicrobial drugs, there has been great interest in the use of metal nanoparticles as coatings for medical devices, wound dressings, and consumer packaging, due to their antimicrobial properties. The wide variety of methods available for nanoparticle synthesis results in a broad spectrum of chemical and physical properties which can affect antibacterial efficacy...
June 2, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605387/a-simple-fractionated-extraction-method-for-the-comprehensive-analysis-of-metabolites-lipids-and-proteins-from-a-single-sample
#15
Mohamed Salem, Michal Bernach, Krzysztof Bajdzienko, Patrick Giavalisco
Understanding of complex biological systems requires the measurement, analysis and integration of multiple compound classes of the living cell, usually determined by transcriptomic, proteomic, metabolomics and lipidomic measurements. In this protocol, we introduce a simple method for the reproducible extraction of metabolites, lipids and proteins from biological tissues using a single aliquot per sample. The extraction method is based on a methyl tert-butyl ether: methanol: water system for liquid: liquid partitioning of hydrophobic and polar metabolites into two immiscible phases along with the precipitation of proteins and other macromolecules as a solid pellet...
June 1, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605384/analyzing-supercomplexes-of-the-mitochondrial-electron-transport-chain-with-native-electrophoresis-in-gel-assays-and-electroelution
#16
Gisela Beutner, George Arthur Porter
The mitochondrial electron transport chain (ETC) transduces the energy derived from the breakdown of various fuels into the bioenergetic currency of the cell, ATP. The ETC is composed of 5 massive protein complexes, which also assemble into supercomplexes called respirasomes (C-I, C-III, and C-IV) and synthasomes (C-V) that increase the efficiency of electron transport and ATP production. Various methods have been used for over 50 years to measure ETC function, but these protocols do not provide information on the assembly of individual complexes and supercomplexes...
June 1, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605370/quantifying-abdominal-pigmentation-in-drosophila-melanogaster
#17
Omid Saleh Ziabari, Alexander W Shingleton
Pigmentation is a morphologically simple but highly variable trait that often has adaptive significance. It has served extensively as a model for understanding the development and evolution of morphological phenotypes. Abdominal pigmentation in Drosophila melanogaster has been particularly useful, allowing researchers to identify the loci that underlie inter- and intraspecific variations in morphology. Hitherto, however, D. melanogaster abdominal pigmentation has been largely assayed qualitatively, through scoring, rather than quantitatively, which limits the forms of statistical analysis that can be applied to pigmentation data...
June 1, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605369/real-time-tracking-of-dna-fragment-separation-by-smartphone
#18
Chunxian Tao, Bo Yang, Zhenqing Li, Dawei Zhang, Yoshinori Yamaguchi
Slab gel electrophoresis (SGE) is the most common method for the separation of DNA fragments; thus, it is broadly applied to the field of biology and others. However, the traditional SGE protocol is quite tedious, and the experiment takes a long time. Moreover, the chemical consumption in SGE experiments is very high. This work proposes a simple method for the separation of DNA fragments based on an SGE chip. The chip is made by an engraving machine. Two plastic sheets are used for the excitation and emission wavelengths of the optical signal...
June 1, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605377/an-anesthesia-surgery-and-harvest-method-for-the-evaluation-of-transpedicular-screws-using-an-in-vivo-porcine-lumbar-spine-model
#19
Young Jae Moon, Jong-Kil Kim, Hong-Geun Oh, Ji-Hun Kang, Gun-Joo Park, Kwang-Bok Lee
Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle screws after spinal surgery, which is a serious concern. To address this problem, diverse types of pedicle screws have been examined to identify those with good fixation strength and osseointegration in spine bone. The porcine spine is a good alternative for the human spine in the evaluation of pedicle screws due to the anatomical size, mechanical characteristics, and cost...
May 31, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28605376/anatomically-inspired-three-dimensional-micro-tissue-engineered-neural-networks-for-nervous-system-reconstruction-modulation-and-modeling
#20
Laura A Struzyna, Dayo O Adewole, Wisberty J Gordián-Vélez, Michael R Grovola, Justin C Burrell, Kritika S Katiyar, Dmitriy Petrov, James P Harris, D Kacy Cullen
Functional recovery rarely occurs following injury or disease-induced degeneration within the central nervous system (CNS) due to the inhibitory environment and the limited capacity for neurogenesis. We are developing a strategy to simultaneously address neuronal and axonal pathway loss within the damaged CNS. This manuscript presents the fabrication protocol for micro-tissue engineered neural networks (micro-TENNs), implantable constructs consisting of neurons and aligned axonal tracts spanning the extracellular matrix (ECM) lumen of a preformed hydrogel cylinder hundreds of microns in diameter that may extend centimeters in length...
May 31, 2017: Journal of Visualized Experiments: JoVE
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