In Vivo Labeling of Cortical Astrocytes with Sulforhodamine 101 (SR101)

Nimmerjahn, Axel; Helmchen, Fritjof

doi:10.1101/pdb.prot068155

Cited by 47 publications

(38 citation statements)

References 29 publications

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“…However, SR101-positive EGFP-negative cells in the hippocampus resemble astrocytes in shape, size and process morphology, corroborating earlier observations by other groups [6], [8]. When we performed whole-cell recordings from SR101-positive EGFP-negative cells in the stratum radiatum of the hippocampus we only found cells that had electrophysiological properties of astrocytes (Figure 2).…”

Section: Discussionsupporting

confidence: 90%

“…Unequivocal identification of astrocytes for imaging experiments as well as electrophysiological recordings was facilitated by the availability of transgenic mouse lines expressing fluorescent proteins under the control of glia specific promoters [4], [5]. Recently, sulforhodamine 101 (SR101) has been used for specific visualization of astrocytes in cortex and hippocampus [6], [7], [8], [9]. In the spinal cord, however, SR101 has been described as unspecific [10] and it was also reported to label oligodendrocytes in the rabbit retina [11].…”

Section: Introductionmentioning

confidence: 99%

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Active Sulforhodamine 101 Uptake into Hippocampal Astrocytes

2012

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Sulforhodamine 101 (SR101) is widely used as a marker of astrocytes. In this study we investigated labeling of astrocytes by SR101 in acute slices from the ventrolateral medulla and the hippocampus of transgenic mice expressing EGFP under the control of the astrocyte-specific human GFAP promoter. While SR101 efficiently and specifically labeled EGFP-expressing astrocytes in hippocampus, we found that the same staining procedure failed to label astrocytes efficiently in the ventrolateral medulla. Although carbenoxolone is able to decrease the SR101-labeling of astrocytes in the hippocampus, it is unlikely that SR101 is taken up via gap-junction hemichannels because mefloquine, a blocker for pannexin and connexin hemichannels, was unable to prevent SR101-labeling of hippocampal astrocytes. However, SR101-labeling of the hippocampal astrocytes was significantly reduced by substrates of organic anion transport polypeptides, including estron-3-sulfate and dehydroepiandrosterone sulfate, suggesting that SR101 is actively transported into hippocampal astrocytes.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Active Sulforhodamine 101 Uptake into Hippocampal Astrocytes

2012

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“…Data were collected from regions exposed to less than 10 seconds of combined bright field and laser excitation to minimize photobleaching. SR101 (5 animals) was topically administered (250-μmol concentration; human dosages not yet determined) 27 to exposed rat brain and tumor tissue. A separate group (5 animals) received an intracarotid injection of SR101 (1 ml of 250-μmol solution).…”

Section: Fluorophoresmentioning

confidence: 99%

“…It has been used extensively to label astrocytes both in vivo and ex vivo. 4,27,28 The detailed mechanisms of its apparent selective fluorescence as well as its toxicity in in vivo use is still under investigation. Selective staining is achieved through local uptake and gap junction-mediated spread of SR101 following its application.…”

Section: Rapid Fluorescent Dyes Suitable For Ex Vivo Imagingmentioning

confidence: 99%

Potential application of a handheld confocal endomicroscope imaging system using a variety of fluorophores in experimental gliomas and normal brain

Martirosyan

Georges

Eschbacher

et al. 2014

FOC

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Object The authors sought to assess the feasibility of a handheld visible-wavelength confocal endomicroscope imaging system (Optiscan 5.1, Optiscan Pty., Ltd.) using a variety of rapid-acting fluorophores to provide histological information on gliomas, tumor margins, and normal brain in animal models. Methods Mice (n = 25) implanted with GL261 cells were used to image fluorescein sodium (FNa), 5-aminolevulinic acid (5-ALA), acridine orange (AO), acriflavine (AF), and cresyl violet (CV). A U251 glioma xenograft model in rats (n = 5) was used to image sulforhodamine 101 (SR101). A swine (n = 3) model with AO was used to identify confocal features of normal brain. Images of normal brain, obvious tumor, and peritumoral zones were collected using the handheld confocal endomicroscope. Histological samples were acquired through biopsies from matched imaging areas. Samples were visualized with a benchtop confocal microscope. Histopathological features in corresponding confocal images and photomicrographs of H & E–stained tissues were reviewed. Results Fluorescence induced by FNa, 5-ALA, AO, AF, CV, and SR101 and detected with the confocal endomicroscope allowed interpretation of histological features. Confocal endomicroscopy revealed satellite tumor cells within peritumoral tissue, a definitive tumor border, and striking fluorescent cellular and subcellular structures. Fluorescence in various tumor regions correlated with standard histology and known tissue architecture. Characteristic features of different areas of normal brain were identified as well. Conclusions Confocal endomicroscopy provided rapid histological information precisely related to the site of microscopic imaging with imaging characteristics of cells related to the unique labeling features of the fluorophores. Although experimental with further clinical trial validation required, these data suggest that intraoperative confocal imaging can help to distinguish normal brain from tumor and tumor margin and may have application in improving intraoperative decisions during resection of brain tumors.

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“…[14] Marking and tracing How do astrocytes interconnect in the brain? [15] Where do free living lobster larvae go after hatching? [16] Figure 2 Examples of dyes manufactured specifically as biomedical reagents.…”

Section: Introductionmentioning

confidence: 99%

Where do dyes go inside living cells? Predicting uptake, intracellular localisation, and accumulation using QSAR models

Horobin

2014

Coloration Technology

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Uptake of dyes into living cells and organisms is of concern to several diverse groups of people. These include those not wishing dyes to enter cells (e.g. manufacturers and users of textile dyes, or laboratory workers using dyes as analytical reagents) and those requiring dye entry (e.g. biologists imaging cell contents, or clinicians using photoactive dyes as antitumour drugs). This diversity results in the need to consider an extremely wide range of dyes -and indeed of cells and organisms. An overview of methods for predicting uptake and intracellular localisation is provided, followed by a more detailed account of the concepts and procedures involved in decision-rule quantitative structure-activity relationship (QSAR) models. Some of these models permit the prediction of which dyes are likely to enter cells, and which dyes will be excluded. Other models predict where internalised dyes will localise within the live cells. Use of QSAR models to understand intracellular accumulation, redistribution, loss from the cell, and metabolic modification of dyes is also considered. In particular, the relationship of such predictions to toxicity is discussed. An extended case example is provided, describing the modelling of dye binding to nucleic acids in single-cell systems. A further case example then illustrates dye localisation in multicellular organisms. Finally, conclusions, critiques, and probable future directions concerning the QSAR modelling approach to dye uptake and localisation are given. A summary of key QSAR decision rules in the form of decision logic tabulations is provided.

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In Vivo Labeling of Cortical Astrocytes with Sulforhodamine 101 (SR101)

Cited by 47 publications

References 29 publications

Active Sulforhodamine 101 Uptake into Hippocampal Astrocytes

Active Sulforhodamine 101 Uptake into Hippocampal Astrocytes

Potential application of a handheld confocal endomicroscope imaging system using a variety of fluorophores in experimental gliomas and normal brain

Where do dyes go inside living cells? Predicting uptake, intracellular localisation, and accumulation using QSAR models

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