Sulforhodamine 101

Unspecific labelling of oligodendrocytes by sulforhodamine 101 depends on astrocytic uptake via the thyroid hormone transporter OATP1C1 (SLCO1C1).[Pubmed:27519929]

Neurosci Lett. 2016 Sep 19;631:13-8.

The red fluorescent dye Sulforhodamine 101 (SR101) is often used as a marker for astrocytes, although variations of the staining protocol have been shown to influence the preferentially labeled cell type. Here we analyzed SR101-labeling of oligodendrocytes in the hippocampal slices preparation of PLP-EGFP mice. Using different staining protocols, we found robust SR101-labeled oligodendrocytes in the CA1 stratum radiatum of the hippocampus. Application of L-thyroxin, which is known to block SR101 transport into astrocytes via competitive inhibition of the multi-specific OATP1C1 (SLCO1C1) transporter, significantly reduced oligodendrocyte labeling. Since OATP1C1 is not expressed in oligodendrocytes, we conclude that oligodendrocyte labeling with SR101 requires SR101-uptake by astrocytes, which then diffuses to oligodendrocytes via heterotypic gap junctions of the pan-glial network. In summary, unequivocal identification of a particular cell type is not possible by SR101 only, hence caution is recommended when using SR101 in future studies.

Sulforhodamine 101, a widely used astrocyte marker, can induce cortical seizure-like activity at concentrations commonly used.[Pubmed:27457281]

Sci Rep. 2016 Jul 26;6:30433.

Sulforhodamine 101 (SR101) is a preferential astrocyte marker widely used in 2-photon microscopy experiments. Here we show, that topical loading of two commonly used SR101 concentrations, 100 muM and 250 muM when incubated for 10 min, can induce seizure-like local field potential (LFP) activity in both anaesthetized and awake mouse sensori-motor cortex. This cortical seizure-like activity develops in less than ten minutes following topical loading, and when applied longer, these neuronal discharges reliably evoke contra-lateral hindlimb muscle contractions. Short duration (<1 min) incubation of 100 muM and 250 muM SR101 or application of lower concentrations 25 muM and 50 muM of SR101, incubated for 30 and 20 min, respectively, did not induce abnormal LFP activity in sensori-motor cortex, but did label astrocytes, and may thus be considered more appropriate concentrations for in vivo astrocyte labeling. In addition to label astrocytes SR101 may, at 100 muM and 250 muM, induce abnormal neuronal activity and interfere with cortical circuit activity. SR101 concentration of 50 muM or lower did not induce abnormal neuronal activity. We advocate that, to label astrocytes with SR101, concentrations no higher than 50 muM should be used for in vivo experiments.

Limitations of Sulforhodamine 101 for Brain Imaging.[Pubmed:28293173]

Front Cell Neurosci. 2017 Feb 28;11:44.

Since 2004, the red fluorescent dye Sulforhodamine 101 (SR101) has been boosting the functional analysis of astrocytes in a functional environment in an unprecedented way. However, two major limitations have been challenging the usefulness of this tool for cellular imaging: (i) SR101 is not as specific for astrocytes as previously reported; and (ii) discoveries of severe excitatory side effects of SR101 are bearing the risk of unwanted alteration of the system of interest. In this article, we summarize the current knowledge about SR101-labeling protocols and discuss the problems that arise from varying of the staining protocols. Furthermore, we provide a testable hypothesis for the observed hyper-excitability that can be observed when using SR101.

Transcranial amelioration of inflammation and cell death after brain injury.[Pubmed:24317693]

Nature. 2014 Jan 9;505(7482):223-8.

Traumatic brain injury (TBI) is increasingly appreciated to be highly prevalent and deleterious to neurological function. At present, no effective treatment options are available, and little is known about the complex cellular response to TBI during its acute phase. To gain insights into TBI pathogenesis, we developed a novel murine closed-skull brain injury model that mirrors some pathological features associated with mild TBI in humans and used long-term intravital microscopy to study the dynamics of the injury response from its inception. Here we demonstrate that acute brain injury induces vascular damage, meningeal cell death, and the generation of reactive oxygen species (ROS) that ultimately breach the glial limitans and promote spread of the injury into the parenchyma. In response, the brain elicits a neuroprotective, purinergic-receptor-dependent inflammatory response characterized by meningeal neutrophil swarming and microglial reconstitution of the damaged glial limitans. We also show that the skull bone is permeable to small-molecular-weight compounds, and use this delivery route to modulate inflammation and therapeutically ameliorate brain injury through transcranial administration of the ROS scavenger, glutathione. Our results shed light on the acute cellular response to TBI and provide a means to locally deliver therapeutic compounds to the site of injury.

Sulforhodamine 101 as a specific marker of astroglia in the neocortex in vivo.[Pubmed:15782150]

Nat Methods. 2004 Oct;1(1):31-7.

Glial cells have been identified as key signaling components in the brain; however, methods to investigate their structure and function in vivo have been lacking. Here, we describe a new, highly selective approach for labeling astrocytes in intact rodent neocortex that allows in vivo imaging using two-photon microscopy. The red fluorescent dye Sulforhodamine 101 (SR101) was specifically taken up by protoplasmic astrocytes after brief exposure to the brain surface. Specificity was confirmed by immunohistochemistry. In addition, SR101 labeled enhanced green fluorescent protein (EGFP)-expressing astrocytes but not microglial cells in transgenic mice. We used SR101 labeling to quantify morphological characteristics of astrocytes and to visualize their close association with the cortical microvasculature. Furthermore, by combining this method with calcium indicator loading of cell populations, we demonstrated distinct calcium dynamics in astroglial and neuronal networks. We expect SR101 staining to become a principal tool for investigating astroglia in vivo.