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Current Protocols in Mouse Biology

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https://www.readbyqxmd.com/read/29261233/evaluation-of-the-estrous-cycle-reproductive-tract-and-mammary-gland-in-female-mice
#1
Justin D Vidal, Adam J Filgo
Evaluation of the female reproductive system is an important part of basic reproductive biology research, toxicology testing, and mutant mouse phenotype assessment. The female reproductive system is dynamic and the onset of puberty and the normal changes observed during estrous cyclicity can create challenges for an investigator. Experimental work in the female mouse requires an understanding of the potential impact of the estrous cycle and tracking normal changes throughout the cycle allows for control of this key variable...
December 20, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/29261232/a-mouse-model-of-west-nile-virus-infection
#2
Jessica B Graham, Jessica L Swarts, Jennifer M Lund
The use of a mouse model to study the breadth of symptoms and disease severity seen in human West Nile virus (WNV) infection can provide insight into the kinetics of the immune response and the specific pathways responsible for control of WNV infection and viral clearance. Here, we provide protocols for performing WNV infection of mice, as well as complete immunophenotyping analysis of the cellular immune response to infection in both the periphery and the central nervous system in a mouse model of WNV infection...
December 20, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/29261231/the-study-of-host-immune-responses-elicited-by-the-model-murine-hookworms-nippostrongylus-brasiliensis-and-heligmosomoides-polygyrus
#3
T Bouchery, B Volpe, K Shah, L Lebon, K Filbey, G LeGros, N Harris
Hookworm infections (Necator americanus or Ancylostoma duodenale) represent a major neglected tropical disease, affecting approximately 700 million people worldwide, and can cause severe morbidity due to the need for these worms to feed on host blood. N. brasiliensis and H. polygrus, both rodent parasites, are the two most commonly employed laboratory models of experimental hookworm infection. Both parasites evoke type 2 immune responses, and their use has been instrumental in generating fundamental insight into the molecular mechanisms of type-2 immunity and for understanding how the immune response can control parasite numbers...
December 20, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/29261230/assessing-sociability-social-memory-and-pup-retrieval-in-mice
#4
Annemarie Zimprich, Jörn Niessing, Lior Cohen, Lillian Garrett, Jan Einicke, Bettina Sperling, Mathias V Schmidt, Sabine M Hölter
Adaptive social behavior is important in mammals, both for the well-being of the individual and for the thriving of the species. Dysfunctions in social behavior occur in many neurodevelopmental and psychiatric diseases, and research into the genetic components of disease-relevant social deficits can open up new avenues for understanding the underlying biological mechanisms and therapeutic interventions. Genetically modified mouse models are particularly useful in this respect, and robust experimental protocols are needed to reliably assess relevant social behavior phenotypes...
December 20, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28884794/pathology-evaluation-of-developmental-phenotypes-in-neonatal-and-juvenile-mice
#5
Brad Bolon, Susan Newbigging, Kelli L Boyd
Necropsy (or autopsy) is the post mortem dissection of an animal to examine and collect organs and tissues in order to understand the effects and causes of disease. The systematic harvesting of samples at necropsy is an essential step in defining the reason for an unexpected death and in characterizing the features (i.e., phenotype) of a newly discovered condition. Phenotypic evaluation of young (neonatal and juvenile) mice emphasizes morphologic (macroscopic and microscopic) techniques and biochemical (clinical chemistry, hematologic, and molecular) analyses...
September 8, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28884793/using-vascular-landmarks-to-orient-3d-optical-coherence-tomography-images-of-the-mouse-eye
#6
Mark P Krebs
Comparing 3D structural information obtained by optical coherence tomography (OCT) requires accurate alignment of images acquired from individual subjects. Despite the widespread use of OCT to image the anterior and posterior mouse eye, few approaches to align the resulting image data have been described, in part due to a lack of well-characterized landmarks that are suitable for alignment. Here, we provide an OCT acquisition and analysis protocol that incorporates the use of the long posterior ciliary arteries as landmarks...
September 8, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28884792/profiling-of-single-cell-transcriptomes
#7
Wanze Chen, Vincent Gardeux, Antonio Meireles-Filho, Bart Deplancke
Complex biological systems are composed of multiple cell types whose transcriptional activity can vary due to differences in cell state, environmental stimulation, or intrinsic programs. Conventional bulk analysis methods capture the average transcriptional programs of the cell population, thus missing the unique cellular signature of each single cell. In recent years, the development of single-cell RNA-sequencing (scRNA-seq) technologies has provided a powerful approach to dissect the cellular heterogeneity of complex biological systems...
September 8, 2017: Current Protocols in Mouse Biology
https://www.readbyqxmd.com/read/28628219/application-of-swath-proteomics-to-mouse-biology
#8
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
#9
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
#10
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
#11
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
#12
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
#13
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
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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