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Current Protocols in Mouse Biology

Marisa A Brake, Lacramioara Ivanciu, Susan A Maroney, Nicolas D Martinez, Alan E Mast, Randal J Westrick
The mammalian blood coagulation system was designed to restrict blood loss due to injury as well as keep the blood fluid within the blood vessels of the organism. Blood coagulation activity in inbred mouse strains varies widely among strains, suggesting that many genomic variants affect hemostasis. Some of these molecules have been discovered and characterized; however, many are still unknown. Genetically modified mouse technologies are providing a plethora of new mouse models for investigating the regulation of blood coagulation...
March 15, 2019: Current Protocols in Mouse Biology
Chih-Wei Ko, Jie Qu, Min Liu, Dennis D Black, Patrick Tso
This protocol provides a comprehensive reference for the evolution of the lymph fistula model, the mechanism of lipid absorption, the detailed procedure for studying lipid absorption using the lymph fistula model, the interpretation of the results, and consideration of the experimental design. The lymph fistula model is an approach to assess the concentration and rate of a range of molecules transported by the lymph by cannulating lymph duct in animals. In this protocol, mice first undergo surgery with the implantation of cannulae in the duodenum and mesenteric lymph duct and are allowed to recover overnight in Bollman restraining cages housed in a temperature-regulated environment...
February 23, 2019: Current Protocols in Mouse Biology
Máximo E Lange, Richard R E Uwiera, G Douglas Inglis
Gnotobiotic mice are an established, robust model utilized in current research to study host-microbiota interactions. For years isolators have been used to rear germ-free and gnotobiotic mice. However, isolators can be costly and the segregation of treatments within the same isolator is problematic. Recently, methodologies for housing germ-free mice in specially designed individually ventilated cages (IVCs) operated under barrier mode have been developed; however, this equipment is costly and its operation in barrier mode for research involving germ-free mice and pathogens is not permissible under modern biosafety and biosecurity regulations...
January 15, 2019: Current Protocols in Mouse Biology
Yong Hong Chen, Megan S Keiser, Beverly L Davidson
Adeno-associated virus (AAV) vectors are exemplary tools for studying gene function in vivo and are particularly favorable for transferring genes of interest into brain tissues. They have shown great promise as a gene therapy vector for preclinical and clinical applications. However, the ability to use this tool is often hampered because the viruses themselves are not readily available. Many methods have been developed for AAV production. Here, we describe a simple method for small- to medium-scale (1012 -1013 viral particles) production of AAV based on Polyethylenimine Max (PEI Max)-mediated triple transfection of HEK 293 cells and purification with iodixanol gradient ultracentrifugation...
December 2018: Current Protocols in Mouse Biology
Yong Hong Chen, Megan S Keiser, Beverly L Davidson
Viral vectors are a promising tool for effective delivery of genetic material into cells. They take advantage of the natural ability of a virus to deliver a genetic payload into cells while being genetically modified such that their ability to replicate is crippled or removed. Here, an updated overview of routinely used viral vectors, including adeno-associated viruses (AAV), retroviruses/lentiviruses, and adenoviruses (Ads), is provided, as well as perspectives on their advantages and disadvantages in research and gene therapy...
December 2018: Current Protocols in Mouse Biology
Megan S Keiser, Yong Hong Chen, Beverly L Davidson
Stereotaxic intracranial injection of viral vectors is a valuable technique to directly deliver genetic material to a specific population of cells in the central nervous system of a mouse model. This enables scientists to test candidate gene therapies or disease modulators that can then provide insight into the pathological mechanisms of disease. In this article, we present a standardized method of conducting intracranial stereotaxic injection of adeno-associated virus into a specific brain region in a mouse model...
November 5, 2018: Current Protocols in Mouse Biology
Andrew Blease, Patricia Das Neves Borges, Marcia Curtinha, Behzad Javaheri, Isabell S von Loga, Ida Parisi, Jadwiga Zarebska, Andrew Pitsillides, Tonia L Vincent, Paul K Potter
With the increasing availability and complexity of mouse models of disease, either spontaneous or induced, there is a concomitant increase in their use in the analysis of pathogenesis. Among such diseases is osteoarthritis, a debilitating disease with few treatment options. While advances in our understanding of the pathogenesis of osteoarthritis has advanced through clinical investigations and genome-wide association studies, there is still a large gap in our knowledge, hindering advances in therapy. Patient samples are available ex vivo, but these are generally in the very late stages of disease...
September 21, 2018: Current Protocols in Mouse Biology
Gregory C Addicks, Phillip Marshall, Bernard J Jasmin, Jean-Marc Renaud, Hongbo Zhang, Keir J Menzies
Muscle function and health progressively deteriorate during the progression of muscle dystrophies. The ability to objectively characterize muscle function and muscle damage is useful not only when comparing variants of dystrophy models, but also for characterizing the effects of interventions aiming to improve or halt the progressive decline of muscle function and muscle health. The protocols in this chapter describe the use of ex vivo eccentric contraction of the diaphragm muscle as a measure of muscle susceptibility to damage...
September 2018: Current Protocols in Mouse Biology
Shuaiyu Wang, Bao Zhang, Gregory C Addicks, Hui Zhang, Keir J Menzies, Hongbo Zhang
Muscle stem cells (MuSCs) are essential for maintaining muscle homeostasis by providing progenitor cells for muscle regeneration after injury and in muscular diseases. MuSC properties dynamically change, reflecting physiology or pathological status. For instance, MuSCs are activated after muscle injury, but become exhausted in late stages of Duchenne Muscular Dystrophy (DMD) disease and senescent during aging. Therefore, characterization of MuSCs, including proliferation, activation, senescence, and apoptosis, etc...
September 2018: Current Protocols in Mouse Biology
Stephan C Collins, Christel Wagner, Léo Gagliardi, Perrine F Kretz, Marie-Christine Fischer, Pascal Kessler, Meghna Kannan, Binnaz Yalcin
In this article, we present a standardized protocol for fast and robust neuroanatomical phenotyping of the adult mouse brain, which complements a previously published article (doi: 10.1002/cpmo.12) in Current Protocols in Mouse Biology. It is aimed at providing an experimental pipeline within an academic research setting from experimental work to data analysis. Our analysis focuses on one single parasagittal plane, covering the majority of brain regions involved in higher order cognitions such as the cortex, hippocampus, and cerebellum, for a total of 166 parameters of area, length, and cell-level measurements in contrast to 78 parameters in our previously published coronal screen...
September 2018: Current Protocols in Mouse Biology
Lucie Crouzier, Tangui Maurice
Here we provide instructions to measure topographic memory in mice using the Hamlet test, a complex environment. The apparatus mimics a small hamlet with a central agora and five houses, which are functionalized since mice can drink, eat pellets, hide within a small maze, run in an activity wheel, or interact with a stranger mouse behind a grid. The houses are interconnected through a network of streets in a five-arm star shape, and a video tracking system takes information from the activity in each house or follows a single mouse by trajectometry...
June 2018: Current Protocols in Mouse Biology
Brian J Nieman, Matthijs C van Eede, Shoshana Spring, Jun Dazai, R Mark Henkelman, Jason P Lerch
This article describes a detailed set of protocols for mouse brain imaging using MRI. We focus primarily on measuring changes in neuroanatomy, and provide both instructions for mouse preparation and details on image acquisition, image processing, and statistics. Practical details as well as theoretical considerations are provided. © 2018 by John Wiley & Sons, Inc.
June 2018: Current Protocols in Mouse Biology
Pilib Ó Broin, Michael V Beckert, Tomohisa Takahashi, Takeshi Izumi, Kenny Ye, Gina Kang, Patricia Pouso, Mackenzie Topolski, Jose L Pena, Noboru Hiroi
Neonatal vocalization is structurally altered in mouse models of autism spectrum disorder (ASD). Our published data showed that pup vocalization, under conditions of maternal separation, contains sequences whose alterations in a genetic mouse model of ASD impair social communication between pups and mothers. We describe details of a method which reveals the statistical structure of call sequences that are functionally critical for optimal maternal care. Entropy analysis determines the degree of non-random call sequencing...
June 2018: Current Protocols in Mouse Biology
Andrew Blease, Thomas Nicol, Sara Falcone, Becky Starbuck, Simon Greenaway, Marie Hutchinson, Paul K Potter
Aging is inevitable, and our society must deal with the consequences: namely, an increased incidence of disease and ill health. Many mouse models of disease are acute or early onset or are induced in young mice, despite the fact that aging is a significant risk factor for a range of significant diseases. To improve modeling of such diseases, we should incorporate aging into our models. Many systems are affected by aging, with a decline in mitochondrial function, an increase in senescence, a loss of resilience, telomere shortening, and a decline in immune function being key factors in the increased susceptibility to disease that is associated with aging...
June 2018: Current Protocols in Mouse Biology
Stacey J Sukoff Rizzo, Laura C Anderson, Torrian L Green, Tracy McGarr, Gaylynn Wells, Shawn S Winter
The relationship between chronological age (lifespan) and biological age (healthspan) varies amongst individuals. Understanding the normal trajectory and characteristic traits of aging mice throughout their lifespan is important for selecting the most reliable and reproducible measures to test hypotheses. The protocols herein describe assays used for aging studies at The Jackson Laboratory's Mouse Neurobehavioral Phenotyping Facility and include assessments of frailty, cognition, and sensory (hearing, vision, olfaction), motor, and fine motor function that can be used for assessing phenotypes in aged mice across their lifespan as well as provide guidance for setting up and validating these behavioral measures...
June 2018: Current Protocols in Mouse Biology
Pierre L Roubertoux, Adeline Ghata, Michèle Carlier
The immaturity at birth and the slowness of ontogenic processes in mice provide the opportunity to measure rates of development. We describe here 18 measures covering the sensorial and motor onset from birth to weaning. The measures are non-invasive, making a follow-up strategy possible. The first basic protocol indicates how to produce mice with known conceptional or chronological age, as the control of the age is a prerequisite to compare rates of development in groups of mice. The second basic protocol describes a set of methods for identifying the pups during a follow-up study...
March 2018: Current Protocols in Mouse Biology
Junzo Kinoshita, Neal S Peachey
Overall retinal function can be monitored by recording the light-evoked response of the eye at the corneal surface. The major components of the electroretinogram (ERG) provide important information regarding the functional status of many retinal cell types including rod photoreceptors, cone photoreceptors, bipolar cells, and the retinal pigment epithelium (RPE). The ERG can be readily recorded from mice, and this unit describes procedures for mouse anesthesia and the use of stimulation and recording procedures for measuring ERGs that reflect the response properties of different retinal cell types...
March 2018: Current Protocols in Mouse Biology
Marina Gertsenstein, Lauryl M J Nutter
Mice carrying patient-associated point mutations are powerful tools to define the causality of single-nucleotide variants to disease states. Epitope tags enable immuno-based studies of genes for which no antibodies are available. These alleles enable detailed and precise developmental, mechanistic, and translational research. The first step in generating these alleles is to identify within the target sequence-the orthologous sequence for point mutations or the N or C terminus for epitope tags-appropriate Cas9 protospacer sequences...
March 2018: Current Protocols in Mouse Biology
Lillian Garrett, Marie-Claire Ung, Tamara Heermann, Kristina M Niedermeier, Sabine Hölter
A better alignment of preclinical and clinical neurobiological measures could help improve neuropsychiatric disease therapeutic development. This unit describes a compendium of hypothesis-driven neuroanatomical phenotyping strategies to be employed in genetic mouse models. Using neuropsychiatric disease-based alterations as a guide, these are histological and immunohistochemical methodologies also applied to human tissue. They include quantification assays of neurochemical-, newly born neuron- and glial-cell markers, synaptic proteins, regional volumetrics, dendritic complexity and spine number as well as an index of excitation/inhibition balance...
March 2018: Current Protocols in Mouse Biology
Devra D Huey, Stefan Niewiesk
The development of humanized mice has become a prominent tool for translational animal studies of human diseases. Here we show how immune deficient mice can be "humanized" by injections of human umbilical cord stem cells. The engraftment of these cells and development into human lymphocytes has been possible because of the development of novel severely immune deficient mouse strains. Here we present proven protocols for the generation and analysis of humanized mice on the NSG mouse background.
March 2018: Current Protocols in Mouse Biology
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