High-resolution in vivo imaging of microglia using a versatile nongenetically encoded marker

Institute of Physiology II, Eberhard Karls University of Tuebingen, Tuebingen, Germany.
European Journal of Immunology (Impact Factor: 4.03). 08/2012; 42(8):2193-6. DOI: 10.1002/eji.201242436
Source: PubMed


Microglial cells are the innate immune cells of the CNS, whose main role is to monitor the integrity of and to react to any disturbances of brain homeostasis. As such, microglial cells are involved in a large number of CNS insults (e.g. acute CNS injury, brain tumors, apoptosis, infection, ischemia, neurodegenerative diseases) and their engagement can be either neurotoxic or neuroprotective [1, 2]. Despite their critical role in ameliorating or exacerbating disease progression, little is known about the in vivo functional properties of these cells.

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    • "In vivo microglial Ca 2+ signaling has been studied so far in healthy young adult mice (Eichhoff et al. 2011; Schwendele et al. 2012). In general, intact microglial cells in the in vivo brain show both agonist-induced and spontaneous changes in [Ca 2+ ] i . "
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    ABSTRACT: Dysregulation of intracellular Ca(2+) homeostasis has been proposed as a common proximal cause of neural dysfunction during aging and Alzheimer's disease (AD). In this context, aberrant Ca(2+) signaling has been viewed as a neuronal phenomenon mostly related to the dysfunction of intracellular Ca(2+) stores. However, recent data suggest that, in AD, Ca(2+) dyshomeostasis is not restricted to neurons but represents a global phenomenon affecting virtually all cells in the brain. AD-related aberrant Ca(2+) signaling in astrocytes and microglia, which is activated during the disease, probably contributes profoundly to an inflammatory response that, in turn, impacts neuronal Ca(2+) homeostasis and brain function. Based on recent data obtained in vivo and in vitro, we propose that bidirectional interactions between the inflammatory responses of glial cells and aberrant Ca(2+) signaling represent a vicious cycle accelerating disease progression.
    Cell and Tissue Research 02/2014; 357(2). DOI:10.1007/s00441-014-1798-8 · 3.57 Impact Factor
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    • "FITC and TRITC dextran [14,56], non-targeted quantum dots [36] and angiosense probes [146] are already available. Injection of fluorescent lectin allows visualization of vessel walls [147] and of fluorophore-conjugated antibodies allows specific vascular subdomains to be highlighted [72,101]. "
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    ABSTRACT: Intravital microscopy has become increasingly popular over the past few decades because it provides high-resolution and real-time information about complex biological processes. Technological advances that allow deeper penetration in live tissues, such as the development of confocal and two-photon microscopy, together with the generation of ever-new fluorophores that facilitate bright labelling of cells and tissue components have made imaging of vertebrate model organisms efficient and highly informative. Genetic manipulation leading to expression of fluorescent proteins is undoubtedly the labelling method of choice and has been used to visualize several cell types in vivo. This approach, however, can be technically challenging and time consuming. Over the years, several dyes have been developed to allow rapid, effective and bright ex vivo labelling of cells for subsequent transplantation and imaging. Here, we review and discuss the advantages and limitations of a number of strategies commonly used to label and track cells at high resolution in vivo in mouse and zebrafish, using fluorescence microscopy. While the quest for the perfect label is far from achieved, current reagents are valuable tools enabling the progress of biological discovery, so long as they are selected and used appropriately.
    Interface focus: a theme supplement of Journal of the Royal Society interface 06/2013; 3(3):20130001. DOI:10.1098/rsfs.2013.0001 · 2.63 Impact Factor
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    ABSTRACT: Here we provide a protocol for rapidly labeling different cell types, distinct subcellular compartments and key injury mediators in the spinal cord of living mice. This method is based on the application of synthetic vital dyes to the surgically exposed spinal cord. Suitable vital dyes applied in appropriate concentrations lead to reliable in vivo labeling, which can be combined with genetic tags and in many cases preserved for postfixation analysis. In combination with in vivo imaging, this approach allows the direct observation of central nervous system physiology and pathophysiology at the cellular, subcellular and functional level. Surgical exposure and preparation of the spinal cord can be achieved in less than 1 h, and then dyes need to be applied for 30-60 min before the labeled spinal cord can be imaged for several hours.
    Nature Protocol 02/2013; 8(3):481-90. DOI:10.1038/nprot.2013.022 · 9.67 Impact Factor
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