Protocols in Current Issue
    Quantification of Densities of Bacterial Endosymbionts of Insects by Real-time PCR
    Author:  Daisuke Kageyama, date: 10/05/2017, view: 220, Q&A: 0
    [Abstract] Increased attention has been paid to the endosymbiotic bacteria of insects. Because most insect endosymbionts are uncultivable, quantitative PCR (qPCR) is a practical and convenient method to quantify endosymbiont titers. Here we report a protocol for real-time qPCR based on SYBR Green I fluorescence as well as some tips to prevent possible ...
    Protease Activity Assay in Fly Intestines
    Authors:  Marie-Paule Nawrot-Esposito, Rihab Loudhaief and Armel Gallet, date: 09/20/2017, view: 457, Q&A: 0
    [Abstract] The intestine is a central organ required for the digestion of food, the absorption of nutrients and for fighting against aggressors ingested along with the food. Impairment of gut physiology following mucosal damages impacts its digestive capacities that consequently will affect growth, wellbeing or even survival of the individual. Hence, the ...
    Drosophila Fecal Sampling
    Authors:  Christine Fink , Jakob von Frieling, Mirjam Knop and Thomas Roeder, date: 09/20/2017, view: 375, Q&A: 0
    [Abstract] Fecal sampling is a non-invasive method which raises the possibility to study the development and the changes in the microbial community throughout different time points of a fly population or throughout different treatments. This method allows precise manipulation to trigger the fly’s physiology by nutritional interventions, bacterial infections ...
    Rearing of Culex spp. and Aedes spp. Mosquitoes
    [Abstract] Mosquito-transmitted pathogens cause major public health problems and contribute substantially to the global burden of disease. Aedes mosquitoes transmit dengue, Zika, yellow fever, and Chikungunya viruses; Culex mosquitoes transmit West Nile, Japanese encephalitis, and Saint Louis encephalitis viruses, among others. Experiments ...
    FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
    Authors:  Nadezhda S. Sabeva and Maria Bykhovskaia, date: 09/05/2017, view: 489, Q&A: 0
    [Abstract] We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are ...
    Cell Type-specific Metabolic Labeling of Proteins with Azidonorleucine in Drosophila
    [Abstract] Advanced mass spectrometry technology has pushed proteomic analyses to the forefront of biological and biomedical research. Limitations of proteomic approaches now often remain with sample preparations rather than with the sensitivity of protein detection. However, deciphering proteomes and their context-dependent dynamics in subgroups of ...
    Preparation of Mosquito Salivary Gland Extract and Intradermal Inoculation of Mice
    Authors:  Michael A. Schmid, Elizabeth Kauffman, Anne Payne, Eva Harris and Laura D. Kramer, date: 07/20/2017, view: 1106, Q&A: 0
    [Abstract] Mosquito-transmitted pathogens are among the leading causes of severe disease and death in humans. Components within the saliva of mosquito vectors facilitate blood feeding, modulate host responses, and allow efficient transmission of pathogens, such as Dengue, Zika, yellow fever, West Nile, Japanese encephalitis, and chikungunya viruses, as well ...
    Plasmodium Sporozoite Motility on Flat Substrates
    Authors:  Henriette L Prinz, Julia M Sattler and Friedrich Frischknecht, date: 07/20/2017, view: 529, Q&A: 0
    [Abstract] Plasmodium sporozoites are the infectious, highly motile forms of the malaria parasite transmitted by Anopheles mosquitoes. Sporozoite motility can be assessed following the dissection of Anopheles salivary glands and isolation of sporozoites in vitro.
    Ciberial Muscle 9 (CM9) Electrophysiological Recordings in Adult Drosophila melanogaster
    Authors:  Benjamin A. Eaton and Rebekah E. Mahoney, date: 07/20/2017, view: 565, Q&A: 0
    [Abstract] The complexity surrounding presynaptic recordings in mammals is a significant barrier to the study of presynaptic mechanisms during neurotransmission in the mammalian central nervous system (CNS). Here we describe an adult fly neuromuscular junction (NMJ), the ciberial muscle 9 (CM9) NMJ, which allows for the recording of both evoked (EPSPs) and ...
    Ex vivo Ooplasmic Extract from Developing Drosophila Oocytes for Quantitative TIRF Microscopy Analysis
    Authors:  Imre Gáspár and Anne Ephrussi, date: 07/05/2017, view: 902, Q&A: 0
    [Abstract] Understanding the dynamic behavior and the continuously changing composition of macromolecular complexes, subcellular structures and organelles is one of areas of active research in both cell and developmental biology, as these changes directly relate to function and subsequently to the development and homeostasis of the organism. Here, we ...