RNA Interference in Ticks

JoVE 2474 1/20/2011

Katherine M. Kocan¹, Edmour Blouin¹, José de la Fuente^1,2

¹Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, ²(CSIC-UCLM-JCCM), Instituto de Investigación en Recursos Cinegéticos IREC

A method for RNA interference (RNAi) by injection of dsRNA into unfed ticks is described. RNAi is the most widely used gene-silencing technique in ticks where the use of other methods of genetic manipulation has been limited.

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

JoVE 3894 2/21/2012

Toni G. Patton¹, Gabrielle Dietrich², Kevin Brandt¹, Marc C. Dolan², Joseph Piesman², Robert D. Gilmore Jr.¹

¹Microbiology and Pathogenesis Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, ²Tick-Borne Diseases Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention

The collection of infected tick hemolymph, salivary glands, and saliva is important to study how tick-borne pathogens cause disease. In this protocol we demonstrate how to collect hemolymph and salivary glands from feeding Ixodes scapularis nymphs. We also demonstrate saliva collection from female I. scapularis adults.

Methods for Rapid Transfer and Localization of Lyme Disease Pathogens Within the Tick Gut

JoVE 2544 2/14/2011

Toru Kariu¹, Adam S. Coleman¹, John F. Anderson², Utpal Pal¹

¹Department of Veterinary Medicine, University of Maryland, ²Department of Entomology, Connecticut Agricultural Experiment Station

Lyme disease research studies often require generation of ticks infected with the pathogen Borrelia burgdorferi, a process that typically takes several weeks. Here we demonstrate a microinjection-based tick infection procedure that can be accomplished within hours. We also demonstrate an immunofluorescence method for in situ localization of B. burgdorferi within ticks.

February 2012: This Month in JoVE

JoVE 4258 2/01/2012

Aaron Kolski-Andreaco

A Calcium Bioluminescence Assay for Functional Analysis of Mosquito (Aedes aegypti) and Tick (Rhipicephalus microplus) G Protein-coupled Receptors

JoVE 2732 4/20/2011

Hsiao-Ling Lu¹, Cymon N. Kersch¹, Suparna Taneja-Bageshwar ^1,2, Patricia V. Pietrantonio¹

¹Department of Entomology, Texas A&M University (TAMU), ²Department of Molecular and Cellular Medicine, Texas A&M University (TAMU)

This protocol provides instructions for clonal-cell line selection and a calcium bioluminescence assay to analyze the structure-activity relationships of synthesized arthropod neuropeptides on their cognate GPCRs. This assay can be used for receptor deorphanization and structure-activity relationship studies for synthetic analog design and peptide/drug-lead discovery.

Clonogenic Assay: Adherent Cells

JoVE 2573 3/13/2011

Haloom Rafehi^1,2, Christian Orlowski^1,2,3, George T. Georgiadis¹, Katherine Ververis^1,4, Assam El-Osta^2,3, Tom C. Karagiannis^1,2

¹Epigenomic Medicine, BakerIDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ²Department of Pathology, The University of Melbourne, ³Epigenetics in Human Health and Disease, BakerIDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ⁴Department of Anatomy and Cellular Biology, The University of Melbourne

The applicability of the clonogenic assay for evaluating reproductive viability has been established for more than 50 years. Here we demonstrate the general procedure for performing the clonogenic assay with adherent cells.

Intracellular Refolding Assay

JoVE 3540 1/24/2012

Tamara Vanessa Walther, Danilo Maddalo

Institute of Toxicology and Genetics, Karlsruhe Institute of Technology

In this protocol a method to measure intracellular protein refolding after heat shock is described. This method can be used to study foldases like molecular chaperones and their co-factors or compounds able to influence their activity. Firefly luciferase activity is used as reporter to measure chaperone refolding activity.

Targeting of Deep Brain Structures with Microinjections for Delivery of Drugs, Viral Vectors, or Cell Transplants

JoVE 2082 12/01/2010

Oscar Gonzalez-Perez¹, Hugo Guerrero-Cazares², Alfredo Quiñones-Hinojosa²

¹ Neuroscience Lab/ Fac. Psicologia, University of Colima, ²Department of Neurosurgery, Johns Hopkins University

In this article, we show a method to make glass capillary needles with a 50-μm lumen. This technique significantly reduces the brain damage, minimizes passive diffusion of drugs and allows a precise targeting into the rodent brain.

Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice

JoVE 2767 9/06/2011

Christina M. Parkinson¹, Alexandra O'Brien¹, Theresa M. Albers¹, Meredith A. Simon¹, Charles B. Clifford¹, Kathleen R. Pritchett-Corning^2,3

¹Research Animal Diagnostic Services, Charles River, ²Research Models and Services, Charles River, ³Department of Comparative Medicine, University of Washington

This article describes various procedures for screening rats and mice to detect endo- or ectoparasitism. Several diagnostic assays will be demonstrated, both those suitable for use on live animals and those used after euthanasia of the animal. Photographs to aid in identification of rat and mouse parasites will be included.

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

JoVE 3397 3/06/2012

Tamalee R. Kramp, Kevin Camphausen

Radiation Oncology Branch, National Cancer Institute

The purpose of this article is to describe the use of an orthotopic glioblastoma model for chemoradiation studies. This article will go though cell processing, implanting, and radiotherapy of the mouse using an intracranial model.