Microglia is a resident macrophage of the parenchyme central nervous system and fulfills the important role in brain development, homeostasis, and inflammation. The pure microglia population isolation of the brain tissue allows microglial phenotypical examinations without other population cell intervalences. The microglial extraction of the neonatal brain has been described in various protocols, but the brain of adult mice that is more established and complex causes a greater challenge.
Here we describe the enhanced protocol including enzymatic and mechanical dissociation of adult mouse brain tissue and the removal of myelin with a gradient of the Percoll density. Mikroglial cells are then extracted by immunomagnetic approaches. This insulation procedure allows functional use of cells for various applications such as cell culture, flow of flow, functional tests including bacteria or bead-based phagocytosis, stimulation tests, and transcript profile techniques such as QRT-PCR and quencing microarray / RNA.
Considerations and guidelines for mouse metabolic phenotypes in diabetes research.
Mice are the most commonly used species in preclinical research on the pathophysiology of metabolic diseases. Although they are very useful for identifying lines, mechanisms and genes that regulate glucose homeostasis and energy, the specificity of various mouse models and methodologies used to investigate metabolic phenotypes can have a profound impact on the experimental and interpretation results. This review aims to: (1) describe the most commonly used experimental tests to assess glucose homeostasis and energy in mice; (2) provide several guidelines for the design, analysis and interpretation of this test, as well as for studies using genetic models; and (3) identify important warnings and confounding factors that must be taken into account in the interpretation of the findings.
The metabolic phenotype in the mouse model of urinary tract infection shows that the 3-hydroxybutyrate in plasma is associated with infection.
Urinary tract infections are one of the most common bacterial infections throughout the world. The current diagnosis of urinary tract infections especially depend on the clinical presentation, urine dipstick tests and urinary culture. Small molecules found in bio fluid related to infection and recovery will facilitate the diagnosis and management of ISK. Fingerprint based on plasma mass spectrometry and urine at 3 point in time, pre-infection (t = -24h), infection (t = 24 hours) and posting 3 days (t = 112h), obtained in the following four groups: healthy mice, Infected but untreated, infected and treated with ciprofloxacin, and infected and treated with Relinqing® granules (n = 6 per group).
Metabolomy workflows include multivariate analysis and ROC regression are used to select metabolic features that correlate with ISK and its maintenance. Features of around 4,000 molecular are obtained for each sample. 3-hydroxybutyrate small acid in plasma found differentiation for urinary tract infections, with the area under curve = 0.97 (confidence interval 95%: 0.93-1.00, accuracy = 0.91, sensitivity = 0.91). The 3-hydroxybutyrate level in plasma runs out after infection with changes to fold -22 (q <0.0001).
The correlation between the 3-hydroxybutyrate plasma and the urine bacterial number in all groups and time points is R = -0.753 (p <0.0001). The findings show that the 3-hydroxybutyrate runs out in the blood and is strongly related to the UTI at the stage of infection and post-treatment in the Mouse model. Jobs are further considered to assess the clinical potential of blood tests to help UTI management.
Understanding the function of genes and mechanisms of the disease: the phenotype pipe at the German mouse clinic.
Since decades, model organisms have provided an important approach to understanding the mechanistic basis of human diseases. The German Mouse Clinic (GMC) is the first phenotype facility to form a collaboration-based platform for the characterization of the phenotype of the mouse lane. To overcome individual projects with specially created phenotyping strategies, GMC advanced in developing a series of pipes with tests for certain disease area analysis.
For general general analysis, there is a screening pipe which includes the main parameters for the most relevant disease area. For phenotypical analysis driven hypotheses, there are thirteen additional pipes with focusing on neurological disorders and behavior, metabolic dysfunction, damage to respiratory systems, immune system disorders and imaging techniques. In this article, we provide an overview of the pipe and illustrate the scientific reasons behind different test combinations.
Various MRI applications for the mouse phenotype.
Models of small animals, especially mouse models, human diseases are very necessary tools for biomedical research. Studies in animal models have provided important insights on the etiology of the disease and accelerating the development of therapeutic strategies. The detailed phenotypic characterization is very important, both for the development of animal models and mechanistic studies into pathogenesis of disease and testing experimental therapeutic efforts.
MRI is multipurpose and non-invasive imaging modalities with excellent penetration depth, tissue coverage, and soft tissue contrast. MRI, is a multi-capital imaging modality, along with a proven imaging protocol and the availability of good contrast agents, ideally suitable for the Mutant Mouse phenotype model. Here we describe the MRI application for phenotypical-birth worms that involve the brain, heart, and kidneys in mice.
The flexibility of MRI and the ease of use is very suitable to meet rapidly increasing demand for the phenotype of the mouse in the age of functional genomics that come.