Current protocols in mouse biology最新文献

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. © 2018 by John Wiley & Sons, Inc.

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. Support protocols are provided for virus dialysis and testing new stereotaxic coordinates with dye. © 2018 by John Wiley & Sons, Inc.

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. However, with mice, we are able to induce disease at a defined time and track the progression in vivo and ex vivo, from inception to end stage, to delineate the processes involved in disease development. © 2018 by John Wiley & Sons, Inc.

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. Because muscle has a robust regenerative capacity, unhealthy muscle may be functionally close to normal; therefore, protocols for ex vivo characterization of muscle are often essential for assessing the effects of interventions. Additional methods that can be applied for assessment of dystrophic muscle are also highlighted. © 2018 by John Wiley & Sons, Inc.

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., is very important in applying MuSC knowledge to regenerative medicine, such as in the treatment of DMD and to improve muscle function in aging. Here, we describe a detailed method for characterizing MuSCs in situ using immunostaining techniques in the mouse. This method can also be easily adapted to analyze other skeletal muscle properties. © 2018 by John Wiley & Sons, Inc.

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. Benefits of using parasagittal analysis for large-scale neuroanatomic screens are discussed. © 2018 by John Wiley & Sons, Inc.

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. Researchers aiming to study aging phenotypes require access to aged mice as a reference when initiating these types of studies in order to observe normal aging characteristics that cannot be observed in young adult mouse populations. © 2018 by John Wiley & Sons, Inc.

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. Training the mice in the Hamlet, in groups for several hours per day over several days or weeks, allows consolidation of topographic memory (i.e., route learning involving both allocentric and egocentric strategies). Analysis of topographic memory can be performed a posteriori in a probe test by depriving mice of water or food and measuring their ability to efficiently reach the “eat” or “drink” house. Control groups include mice tested in non-deprived condition and mice naïve to the Hamlet and tested in deprived or non-deprived conditions. The present article details the apparatus, procedures, and protocols that can be used to reliably habituate mice in this complex environment and measure topographic memory. © 2018 by John Wiley & Sons, Inc.

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.