quantifications of western blots with imagej

Infiltrating macrophages/neutrophils also contribute to inflammatory responses in the brain however, the timing of their arrival, numbers, and distinct functions are often highly debated. Microglia are the resident brain immune cells and a key player in the neuroinflammatory response following an ischemic stroke. While potent neuroinflammatory responses can exacerbate injury in the acute phase and, recently discovered, are slow to resolve in the brain, neuroinflammatory and microglial responses also contribute to brain recovery by influencing brain remodeling and repair. Post-stroke neuroinflammation includes a complex, multi-faceted, and often sex-specific cellular response to injury that contributes to brain recovery and repair after ischemic stroke. As such, biological sex and menopause status are important variables to consider in stroke studies that investigate gender-based differences. Males have a higher age-specific incidence of ischemic stroke than women however, women experience an increase in stroke severity and incidence postmenopause. In the USA, the age-standardized prevalence of ischemic stroke is among the highest in the world and a significant cause of mortality and morbidity. Ischemic stroke is a global concern as the burden of stroke continues to rise with few effective prevention strategies or treatments on the horizon. Until TMEM119 function in the brain is elucidated, its use to distinguish between cell populations following brain injury with cell infiltration is cautioned. Our findings suggest that TMEM119 is not a stable microglia marker in male and female mice in the context of ischemic stroke. Second, whereas the TMEM119 immunofluorescence area decreased in proximity to the infarcted area, the TMEM119 protein quantity was unchanged in the ipsilateral hemisphere regions using western blot methods. Cells with severely decreased TMEM119 immunofluorescence were ramified, a distinguishing microglia characteristic. First, morphological data were not congruent with TMEM119 immunofluorescence data. Two key findings prevented distinguishing microglia from systemic macrophages. The number of TMEM119-positive cells was decreased in proximity to the infarct ( p < 0.0001) but without a sex group effect. However, the number of phagocytic microglia was increased in distal ipsilateral regions of postmenopausal mice vs. After stroke, the microglia underwent a region, but not sex group, dependent transformation into less ramified cells ( p < 0.0001). premenopause female mice ( p < 0.05) with differences primarily occurring in the caudal sections. Post-stroke injury is increased in male and postmenopause female mice vs. Western blot was used to quantify protein quantity. Ionized calcium-binding adapter molecule, TMEM119, and confocal microscopy were used to analyze the microglia morphology and TMEM119 area in the ipsilateral brain regions. Brain tissue was collected to quantify the infarct area and for immunohistochemistry and western blot methods. The accelerated ovarian failure model was used to model postmenopause. Male and female, premenopausal and postmenopausal, mice underwent either 60 min of middle cerebral artery occlusion and 24 h of reperfusion or sham surgery. Second, we examined sex and menopause differences in microglia/macrophage cell populations after an ischemic stroke. Our purpose was to identify microglial and macrophage populations regulated by ischemic stroke using morphology analysis and the presence of microglia transmembrane protein 119 (TMEM119). A persistent obstacle in understanding the sex-specific neuroinflammatory contributions to ischemic brain injury is distinguishing between resident microglia and infiltrating macrophages-both phagocytes-and determining cell population-specific contributions to injury evolution and recovery processes. Ischemic stroke is an acquired brain injury with gender-dependent outcomes.

Quantifications of western blots with imagej