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Current Protocols in Mouse Biology

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https://www.readbyqxmd.com/read/28628219/application-of-swath-proteomics-to-mouse-biology
#1
Yibo Wu, Evan G Williams, Ruedi Aebersold
The quantitative measurement of the proteome has been shown to yield new insights into physiology and cell biology that cannot be determined from the genome and transcriptome because the quantitative relationship between transcriptome and proteome is complex. MS-based proteomics techniques, such as SWATH-MS, have recently advanced to the point at which they may be reliably applied by biologists who are not specialists in mass spectrometry. Here we provide standard protocols for preparation of tissue samples for input into the SWATH-MS analytical pipeline...
June 19, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28628218/microbiota-analysis-using-an-illumina-miseq-platform-to-sequence-16s-rrna-genes
#2
Alexis Rapin, Céline Pattaroni, Benjamin J Marsland, Nicola L Harris
The microbiota have been shown to play an important role in diverse biological processes including immunity, metabolism, and digestion. Assessing the exact composition of the microbiota has proven challenging due to the often unknown growth specificities of its members, and culture-based approaches typically fail to capture the complete diversity of microorganisms present. Next Generation Sequencing (NGS) methods provide an efficient means to gather information about cultured and uncultured members of the microbiota...
June 19, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28628217/mouse-model-of-burn-wound-and-infection-thermal-hot-air-lesion-induced-immunosuppression
#3
Henrik Calum, Niels Høiby, Claus Moser
The immunosuppression induced by thermal injury renders the burned victim susceptible to infection. A mouse model was developed to examine the immunosuppression, which was possible to induce even at a minor thermal insult of 6% total body surface area. After induction of the burn (48 hr) a depression of leukocytes in the peripheral blood was found of the burned mice. This depression was due to a reduction in the polymorphonuclear cells. The burned mice were not able to clear a Pseudomonas aeruginosa wound infection, since the infection spread to the blood as compared to mice only infected with P...
June 19, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28628216/a-fast-easy-and-customizable-eight-color-flow-cytometric-method-for-analysis-of-the-cellular-content-of-bronchoalveolar-lavage-fluid-in-the-mouse
#4
François Daubeuf, Julien Becker, Juan Antonio Aguilar-Pimentel, Claudine Ebel, Martin Hrabě de Angelis, Yann Hérault, Nelly Frossard
The cell composition of bronchoalveolar lavage fluid (BAL) is an important indicator of airway inflammation. It is commonly determined by cytocentrifuging leukocytes on slides, then staining, identifying, and counting them as eosinophils, neutrophils, macrophages, or lymphocytes according to morphological criteria under light microscopy, where it is not always easy to distinguish macrophages from lymphocytes. We describe here a one-step, easy-to-use, and easy-to-customize 8-color flow cytometric method for performing differential cell count and comparing it to morphological counts on stained cytospins...
June 19, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28628215/monitoring-pathogen-induced-sickness-in-mice-and-rats
#5
Daria Kolmogorova, Emma Murray, Nafissa Ismail
Sickness behavior monitoring, a technique for examining the development of sickness symptomatology following infection, is necessary in experiments studying neurochemical and physiological changes associated with pathogen-induced immune activation. However, the results of sickness behavior monitoring are difficult to reconcile due to inconsistencies in protocol methods and rater bias. The protocol described herein offers a non-invasive and unbiased approach to assess the progression of pathogen-induced sickness behaviors...
June 19, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28252202/a-mouse-model-for-ocular-surface-staphylococcus-aureus-infection
#6
Zhiyong Zhang, Osama Abdel-Razek, Guirong Wang
Creation of an appropriate animal model that accurately reflects the disease and host immune response to bacterial infection in humans is a major challenge in ocular-surface infection research. For decades, mice have been the ideal small animal model for ocular-surface infection research because of the availability and relatively low cost of various genetic backgrounds, targeted defects, and immunologic reagents. By employing different combinations of mouse and bacterial strains, murine infection models can be used to explore a complete picture of bacterial infection and innate immunity of the ocular surface...
March 2, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28252201/non-typeable-haemophilus-influenzae-infection-of-the-junbo-mouse
#7
Michael T Cheeseman, Derek W Hood
Acute otitis media, inflammation of the middle ear bulla, is the most common bacterial infection in children. For one of the principal otopathogens, non-typeable Haemophilus influenzae (NTHi), animal models allow us to investigate host-microbial interactions relevant to the onset and progression of infection and to study treatment of middle ear disease. We have established a robust model of NTHi middle ear infection in the Junbo mouse. Intranasal inoculation with NTHi produces high rates of bulla infection and high bacterial titers in bulla fluids; bacteria can also spread down the respiratory tract to the mouse lung...
March 2, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28252200/exploration-of-inflammatory-bowel-disease-in-mice-chemically-induced-murine-models-of-inflammatory-bowel-disease-ibd
#8
Raffaella Maria Gadaleta, Oihane Garcia-Irigoyen, Antonio Moschetta
Inflammatory bowel disease (IBD) is a chronic multifactorial inflammatory disorder characterized by periods of activation and remission of intestinal inflammation, with potentially severe complications, that can lead to mortality. Experimental animal models of intestinal inflammation are crucial for understanding the pathogenesis of Crohn's disease (CD) and ulcerative colitis (UC), the two major human IBD phenotypes. Animal models have been instrumental in unveiling the molecular background of IBD, and although a single model is not able to capture the complexity of this disease, each of them provided valuable insight into its different aspects...
March 2, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28252199/analysis-of-mtdna-ndna-ratio-in-mice
#9
Pedro M Quiros, Aashima Goyal, Pooja Jha, Johan Auwerx
Mitochondrial DNA (mtDNA) lacks the protection provided by the nucleosomes in the nuclear DNA and does not have a DNA repair mechanism, making it highly susceptible to damage, which can lead to mtDNA depletion. mtDNA depletion compromises the efficient function of cells and tissues and thus impacts negatively on health. Here, we describe a brief and easy protocol to quantify mtDNA copy number by determining the mtDNA/nDNA ratio. The procedure has been validated using a cohort of young and aged mice. © 2017 by John Wiley & Sons, Inc...
March 2, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28252198/cre-loxp-mediated-recombination-general-principles-and-experimental-considerations
#10
Micheal A McLellan, Nadia A Rosenthal, Alexander R Pinto
The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. Use of the system has greatly expanded our ability to precisely interrogate gene function in the mouse, providing both spatial and temporal control of gene expression. This has been largely due to the simplicity of its use and its adaptability to address diverse biological questions. While the use of the cre-loxP system is becoming increasingly widespread, in particular because of growing availability of conditional mouse mutants, many considerations need to be taken into account when utilizing the cre-loxP system...
March 2, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27906464/methodological-considerations-for-optimizing-and-validating-behavioral-assays
#11
Stacey J Sukoff Rizzo, Jill L Silverman
Preclinical animal models are indispensable tools for translational research for which behavioral characterization and phenotyping are essential to testing hypotheses and for evaluating the potential of novel therapeutic agents to treat diseases. The methods employed for comprehensive behavioral phenotyping and pharmacological experiments are complex and should be conducted exclusively by trained technicians with demonstrated proficiency. The ultimate goal is to identify disease-relevant and translational behavioral endpoints that are robust, reliable, and reproducible, and that can be employed to evaluate potential of novel therapeutic agents to treat disease...
December 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27906463/measurement-of-behavioral-taste-responses-in-mice-two-bottle-preference-lickometer-and-conditioned-taste-aversion-tests
#12
Dany Gaillard, Jennifer M Stratford
The natural like and dislike of foods based on taste is one of the most easily observed behaviors in animals. Animals eat palatable foods and reject aversive foods, which makes measurement of taste perception possible using various behavioral techniques. Three different methods to accurately measure taste behavior are described here. First, two-bottle preference tests evaluate whether a taste compound (tastant) is preferred over water. Second, lickometer tests quantify the like and dislike for multiple concentrations of the same tastant or multiple tastants at the same time...
December 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27906462/histopathological-evaluation-of-skeletal-muscle-with-specific-reference-to-mouse-models-of-muscular-dystrophy
#13
Rebecca L Terry, Dominic J Wells
The muscular dystrophies are a diverse group of degenerative diseases for which many mouse models are available. These models are frequently used to assess potential therapeutic interventions and histological evaluation of multiple muscles is an important part of this assessment. Histological evaluation is especially useful when combined with tests of muscle function. This unit describes a protocol for necropsy, processing, cryosectioning, and histopathological evaluation of murine skeletal muscles, which is applicable to both models of muscular dystrophy and other neuromuscular conditions...
December 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27906461/measurement-of-intestinal-and-peripheral-cholesterol-fluxes-by-a-dual-tracer-balance-method
#14
Onne A H O Ronda, Theo H van Dijk, H J Verkade, Albert K Groen
Long-term elevated plasma cholesterol levels put individuals at risk for developing atherosclerosis. Plasma cholesterol levels are determined by the balance between cholesterol input and output fluxes. Here we describe in detail the methodology to determine the different cholesterol fluxes in mice. The percentage of absorbed cholesterol is calculated from a stable isotope-based double-label method. Cholesterol synthesis is calculated from MIDA after (13) C-acetate enrichment. Cholesterol is removed from the body via the feces...
December 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27584556/metabolic-profiling-and-quantification-of-neurotransmitters-in-mouse-brain-by-gas-chromatography-mass-spectrometry
#15
Christian Jäger, Karsten Hiller, Manuel Buttini
Metabolites are key mediators of cellular functions, and have emerged as important modulators in a variety of diseases. Recent developments in translational biomedicine have highlighted the importance of not looking at just one disease marker or disease inducing molecule, but at populations thereof to gain a global understanding of cellular function in health and disease. The goal of metabolomics is the systematic identification and quantification of metabolite populations. One of the most pressing issues of our times is the understanding of normal and diseased nervous tissue functions...
September 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27584555/histomorphological-phenotyping-of-the-adult-mouse-brain
#16
Anna Mikhaleva, Meghna Kannan, Christel Wagner, Binnaz Yalcin
This article describes a series of standard operating procedures for morphological phenotyping of the mouse brain using basic histology. Many histological studies of the mouse brain use qualitative approaches based on what the human eye can detect. Consequently, some phenotypic information may be missed. Here we describe a quantitative approach for the assessment of brain morphology that is simple and robust. A total of 78 measurements are made throughout the brain at specific and well-defined regions, including the cortex, the hippocampus, and the cerebellum...
September 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27584554/light-and-electron-microscopy-methods-for-examination-of-cochlear-morphology-in-mouse-models-of-deafness
#17
Andrew Parker, Lauren Chessum, Philomena Mburu, Jeremy Sanderson, Michael R Bowl
Mice are an invaluable model organism for the study of auditory function. Even though there are differences in size and frequency response, the anatomy and physiology of the mouse and human ear are remarkably similar. In addition, the tools available for genetic manipulation in the mouse have enabled the generation of models carrying mutations in orthologous human deafness-causing genes, helping to validate these lesions and assess their functional consequence. Reciprocally, novel gene mutations discovered to cause auditory deficits in the mouse highlight potential new loci for human hearing loss, and expand our basic knowledge of the mechanisms and pathways important for the function of the mammalian ear...
September 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27584553/islet-insulin-secretion-measurements-in-the-mouse
#18
Alison Hugill, Kenju Shimomura, Roger D Cox
This article describes detailed protocols for in vitro measurements of insulin function and secretion in isolated mouse islets for the analysis of glucose homeostasis. We specify a method of enzyme digestion and hand picking to isolate and release the greatest number of high quality islets from the pancreas of the mouse. We describe an effective method for generating dynamic measurements of insulin secretion using a perifusion assay including a detailed protocol for constructing a peristaltic pump and tubing assembly...
September 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27584552/peripheral-neuropathy-in-mouse-models-of-diabetes
#19
REVIEW
Corinne G Jolivalt, Katie E Frizzi, Lucie Guernsey, Alex Marquez, Joseline Ochoa, Maria Rodriguez, Nigel A Calcutt
Peripheral neuropathy is a frequent complication of chronic diabetes that most commonly presents as a distal degenerative polyneuropathy with sensory loss. Around 20% to 30% of such patients may also experience neuropathic pain. The underlying pathogenic mechanisms are uncertain, and therapeutic options are limited. Rodent models of diabetes have been used for more than 40 years to study neuropathy and evaluate potential therapies. For much of this period, streptozotocin-diabetic rats were the model of choice...
September 1, 2016: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/27584551/comprehensive-energy-balance-measurements-in-mice
#20
Lee Moir, Liz Bentley, Roger D Cox
In mice with altered body composition, establishing whether it is food intake or energy expenditure, or both, that is the major determinant resulting in changed energy balance is important. In order to ascertain where the imbalance is, the acquisition of reproducible data is critical. Therefore, here we provide detailed descriptions of how to determine energy balance in mice. This encompasses protocols for establishing energy intake from home cage measurement of food intake, determining energy lost in feces using bomb calorimetry, and using equations to calculate parameters such as energy intake (EI), digested energy intake (DEI), and metabolisable energy intake (MEI) to determine overall energy balance...
September 1, 2016: Current Protocols in Mouse Biology
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