Polish Academy of Sciences View Institution's Website 14 articles published in JoVE Neuroscience Motion-Acuity Test for Visual Field Acuity Measurement with Motion-Defined Shapes Marco Ninghetto1, Michał Wieteska2,3, Anna Kozak1, Kamil Szulborski4, Tomasz Gałecki4, Jacek Szaflik4, Kalina Burnat1 1Nencki Institute of Experimental Biology, Polish Academy of Sciences, 2Mossakowski Medical Research Institute, Polish Academy of Sciences, 3Institute of Radio Electronics and Multimedia Technology, Warsaw University of Technology, 4Department of Ophthalmology, Medical University of Warsaw A novel motion-based acuity test that allows the assessment of central and peripheral visual processing in low-vision and healthy individuals, along with goggles limiting peripheral vision compatible with MRI protocols, are described here. This method offers a comprehensive vision assessment for functional impairments and dysfunctions of the visual system. Biochemistry High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution Lukasz S. Borowski1,2, Karolina Kasztelan2,3, Jolanta Czerwinska-Kostrzewska1,2, Roman J. Szczesny2 1Faculty of Biology, Institute of Genetics and Biotechnology, University of Warsaw, 2Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 3International Institute of Molecular and Cell Biology in Warsaw A procedure for studying the dynamics of mitochondrial DNA (mtDNA) metabolism in cells using a multi-well plate format and automated immunofluorescence imaging to detect and quantify mtDNA synthesis and distribution is described. This can be further used to investigate the effects of various inhibitors, cellular stresses, and gene silencing on mtDNA metabolism. Neuroscience Studying Pre-formed Fibril Induced α-Synuclein Accumulation in Primary Embryonic Mouse Midbrain Dopamine Neurons Safak Er1, Irena Hlushchuk1, Mikko Airavaara2, Piotr Chmielarz1,3, Andrii Domanskyi1 1Institute of Biotechnology, HiLIFE, University of Helsinki, 2Neuroscience Center, HiLIFE, University of Helsinki, 3Department of Brain Biochemistry, Maj Institute of Pharmacology, Polish Academy of Sciences Here, we present a detailed protocol to study neuronal α-synuclein accumulation in primary mouse dopamine neurons. Phosphorylated α-synuclein aggregates in neurons are induced with pre-formed α-synuclein fibrils. Automated imaging of immunofluorescently labeled cells and unbiased image analysis make this robust protocol suitable for medium-to-high throughput screening of drugs that inhibit α-synuclein accumulation. Bioengineering Generation of Transgenic Rats using a Lentiviral Vector Approach Paulina Koza1, Joanna Przybyś1, Agata Klejman1, Gabriela Olech-Kochańczyk1, Witold Konopka1 1Nencki Institute of Experimental Biology, Polish Academy of Sciences This article aims to provide the methodology for lentiviral transgenesis in rat embryos using multiple injections of a virus suspension into the zygote perivitelline space. Female rats that are mated with a fertile male strain with a different dominant fur color is used to generate pseudopregnant foster mothers. Developmental Biology Neurogenesis Using P19 Embryonal Carcinoma Cells Paweł Leszczyński1, Magdalena Śmiech1, Aamir S. Teeli1, Aleksandra Zołocińska2, Anna Słysz2, Zygmunt Pojda2, Mariusz Pierzchała3, Hiroaki Taniguchi1 1Department of Experimental Embryology, Institute of Genetics and Animal Breeding, Polish Academy of Sciences, 2Department of Regenerative Medicine, Maria Skłodowska-Curie Institute - Oncology Center, 3Department of Genomics and Biodiversities, Institute of Genetics and Animal Breeding, Polish Academy of Sciences The P19 mouse embryonic carcinoma cell line (P19 cell line) is widely used for studying the molecular mechanism of neurogenesis with great simplification compared to in vivo analysis. Here, we present a protocol for retinoic acid-induced neurogenesis in the P19 cell line. Chemistry Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera Piotr Pander1, Przemyslaw Data1,2,3, Fernando B. Dias1 1Department of Physics, University of Durham, 2Faculty of Chemistry, Silesian University of Technology, 3Center of Polymer and Carbon Materials, Polish Academy of Sciences Here, we present a method of the spectroscopic characterization of organic molecules by means of time-resolved photoluminescence spectroscopy on the nanosecond-to-millisecond timescale in oxygen-free conditions. Methods to efficiently remove oxygen from the samples and, thus, limit luminescence quenching are also described. Engineering Production and Characterization of Vacuum Deposited Organic Light Emitting Diodes Daniel de Sa Pereira1, Andrew P. Monkman1, Przemyslaw Data1,2,3 1Physics Department, Durham University, 2Faculty of Chemistry, Silesian University of Technology, 3Centre of Polymer and Carbon Materials, Polish Academy of Science A protocol for the production of simple structured organic light-emitting diodes (OLEDs) is presented. Biochemistry Manipulating Living Cells to Construct Stable 3D Cellular Assembly Without Artificial Scaffold Takehiro Yamazaki*1, Hiroaki Taniguchi*2, Shoto Tsuji1, Shiho Sato1, Takahiro Kenmotsu1, Kenichi Yoshikawa1, Koichiro Sadakane1 1Faculty of Life and Medical Sciences, Doshisha University, 2The Institute of Genetics and Animal Breeding, Polish Academy of Sciences We demonstrate a novel method for constructing a single-cell-based 3-dimensional (3D) assembly without an artificial scaffold. Medicine An In Vivo Method for Evaluating the Gut-Blood Barrier and Liver Metabolism of Microbiota Products Kinga Jaworska1, Tomasz Huc1, Marta Gawrys1, Maksymilian Onyszkiewicz1, Emilia Samborowska2, Marcin Ufnal1 1Department of Experimental Physiology and Pathophysiology, Laboratory of Centre for Preclinical Research, Medical University of Warsaw, 2Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics, Polish Academy of Sciences The access of nutrients, microbiota metabolites and medicines to the circulation is controlled by the gut-blood barrier (GBB). We describe a direct method for measuring the GBB permeability in vivo, which, in contrast to commonly used indirect methods, is virtually not affected by liver and kidney functions. Developmental Biology Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout (Oncorhynchus Mykiss) and Brown Trout (Salmo Trutta Morpha fario) Radosław Kajetan Kowalski1, Beata Sarosiek1, Sylwia Judycka1, Katarzyna Dryl1, Joanna Grudniewska2, Stefan Dobosz2, Beata Irena Cejko1 1Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Science, 2Department of Salmonid Fish Research, The Stanisław Sakowicz Inland Fisheries Institute The principal aim of this study is to standardize and test the pneumatic method (air stripping) of collecting eggs in rainbow trout and brown trout. This method allows effective and simple collection of the eggs without the necessity of fish abdomen massage. Biochemistry A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis Megumi Tsuchiya*1, M. Rezaul Karim*2, Taro Matsumoto3, Hidesato Ogawa1, Hiroaki Taniguchi3,4,5 1Graduate School of Frontier Biosciences, Osaka University, 2Department of Biotechnology and Genetic Engineering, Jahangirnagar University, 3Division of Cell Regeneration and Transplantation, School of Medicine, Nihon University, 4Institute of Genetics and Animal Breeding of the Polish Academy of Sciences, 5Graduate School of Life and Medical Sciences, Doshisha University We have established a method for the purification of coregulatory interaction proteins using the LC-MS/MS system. Developmental Biology Imaging of Cell Shape Alteration and Cell Movement in Drosophila Gastrulation Using DE-cadherin Reporter Transgenic Flies M. Rezaul Karim1, Tomohiro Haruta2, Taro Matsumoto3, Hiroki Oda2, Hiroaki Taniguchi3,4,5 1Biotechnology and Genetic Engineering Department, Jahangirnagar University, 2JT Biohistory Research Hall, 3Division of Cell Regeneration and Transplantation, School of Medicine, Nihon University, 4Graduate School of Life and Medical Sciences, Doshisha University, 5Institute of Genetics and Animal Breeding of the Polish Academy of Sciences Herein we describe a procedure to capture live images of Drosophila gastrulation. This has enabled us to better understand the apical constriction involved in early development and further analyze mechanisms governing cellular movements during tissue structure modification. Neuroscience A Simple Flight Mill for the Study of Tethered Flight in Insects Alfredo Attisano1, James T. Murphy2, Andrew Vickers3, Patricia J. Moore2 1Museum and Institute of Zoology, Polish Academy of Sciences, 2Department of Entomology, University of Georgia, 3Riventa Pool Innovation Centre Flight in insects is influenced by a number of factors and the propensity to disperse is an important variable in understanding insect ecology and biological control strategies. We describe the construction and use of a simple, relatively inexpensive, and flexible flight mill for measuring parameters of tethered flight in insects. Neuroscience Ex vivo Live Imaging of Single Cell Divisions in Mouse Neuroepithelium Karolina Piotrowska-Nitsche1,2, Tamara Caspary1 1Department of Human Genetics, Emory University School of Medicine, 2Department of Experimental Embryology, IGAB Polish Academy of Sciences Here we develop the tools necessary for ex vivo live imaging to trace single cell divisions in the mouse E8.5 neuroepithelium
