Vivo (2020)
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Microglia are specialized resident macrophages of the central nervous system (CNS) that have important functions during neurodevelopment, homeostasis and disease. This mini-review provides an overview of the current tools and approaches for studying microglia in vivo. We focus on tools for labeling microglia, highlighting the advantages and limitations of microglia markers/antibodies and reporter mice. We also discuss techniques for imaging microglia in situ, including in vivo live imaging of brain and retinal microglia. Finally, we review microglia depletion approaches and their use to investigate microglial function in CNS homeostasis and disease.
Figure 1. (A) Tools and approaches for studying microglia in vivo. (B) Examples of advances in microglia research that have been made using these tools. AD, Alzheimer's disease; AO, adaptive optics; CSF1R, colony stimulating factor 1 receptor; DT, diphtheria toxin; DTR, diphtheria toxin receptor; MG, microglia; SLO, scanning laser ophthalmoscopy; TPEFM, two-photon excitation fluorescence microscopy. *can also be used for in vivo retinal microglia dynamic behavior studies.
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In vivo imaging of tumor hypoxia and angiogenesis with positron emission tomography (PET) is playing an increasingly important role in the diagnosis of tumors. In addition, more effective antitumor treatments can be planned by using new, specific radiopharmaceuticals that detect angiogenesis and hypoxia in malignant tumors. Radiopharmacons that are labelled with positron emitting radionuclides (11C, 18F, and 68Ga) are used in PET imaging wherewith the uptake and the biodistribution of the labelled molecule can be detected and quantificated in vivo [20]. The most commonly used PET radiopharmaceuticals are 18F-FDG, 11C-methionine, and 18F-FLT which give information about cell metabolism, but they are not specific for hypoxia or proteins and receptors that are overexpressed in tumor-associated neoangiogenesis.
In this present study, we hypothesized that the expression of APN/CD13 and the development of hypoxia vary during the growth of subcutaneous hepatocellular carcinoma (He/De) in rats. The aim of this study was to evaluate whether 68Ga-NOTA-c(NGR) and 68Ga-DOTA-nitroimidazole allow the in vivo noninvasive detection of the temporal changes of APN/CD13 expression and hypoxia in experimental He/De tumors using positron emission tomography.
Tumor enlargement is greatly influenced by the oxygen and nutrient supply. In case of reduced oxygen supply (hypoxia), HIF transcriptional factors are activated which promotes gene expression of those responsible for tumor survival and progression. One of these hypoxia and HIF-induced processes is tumor neoangiogenesis [43]. In this present study, for the in vivo assessment of the correspondence between angiogenesis and tumor growth, 68Ga-NOTA-c(NGR) was used due to the fact that the expression of APN/CD13 was higher than that of αvβ3 integrin in He/De tumors. We hypothesized that the hypoxic and angiogenic areas increase in size with tumor volume. This assumption was confirmed since 68Ga-NOTA-c(NGR) and 68Ga-DOTA-nitroimidazole uptake increased and showed a strong correlation with the tumor volume enlargement (Figure 4 and Supplementary data 3: Table 1). Furthermore, we hypothesized that elevation of hypoxia will induce increasingly strong angiogenesis with tumor growth. This was confirmed by indirect evidence that the increasing uptake of 68Ga-DOTA-nitroimidazole was strongly correlated with the accumulation of 68Ga-NOTA-c(NGR) (Fig
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