Dual reporter approaches for identification of Cre efficacy and astrocyte heterogeneity

Degen, Joachim; Dublin, Pavel; Zhang, Jiong; Dobrowolski, Radoslaw; Jokwitz, Melanie; Karram, Khalad; Trotter, Jacqueline; Jabs, Ronald; Willecke, Klaus; Steinhäuser, Christian; Theis, Martin

doi:10.1096/fj.12-207183

Cited by 30 publications

(44 citation statements)

References 41 publications

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“…To investigate the relationship between neuronal and astrocytic calcium, we performed high temporal and spatial resolution imaging and AM in mice expressing enhanced cyan fluorescent protein (ECFP) under control of the astrocyte-specific connexin 43 promoter (Cx43-ECFP) (13). Blood vessels were labeled with Texas Red dextran (Supplemental Figure 2, A and B, and Supplemental Video 1).…”

Section: Resultsmentioning

confidence: 99%

Astrocytic calcium release mediates peri-infarct depolarizations in a rodent stroke model

Rakers¹,

Petzold²

2016

Journal of Clinical Investigation

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Section: Resultsmentioning

confidence: 99%

Astrocytic calcium release mediates peri-infarct depolarizations in a rodent stroke model

Rakers¹,

Petzold²

2016

Journal of Clinical Investigation

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“…Previous work in this field focused on analyzing defined groups of astrocytes filled with different fluorescent dyes [11], [13], [37] or genetically labeled by expression of fluorescent proteins [12], [38], [39]. Both approaches allow exact analysis of the intimate interaction of a limited number of cells.…”

Section: Discussionmentioning

confidence: 99%

“…It is unknown whether the dye is evenly distributed and reaches all the finest processes and spongiform elaborations of protoplasmic astrocytes. Moreover, since astrocytes differ in size, morphology, and gene expression [38], [40], [42]–[46], genetic approaches are limited by the availability of promoters driving the expression of reporter proteins in specific cell types. For example, mice expressing enhanced green fluorescent protein under the control of the GFAP promoter might be an imperfect tool to determine the morphology of astrocytes in healthy cortex, as only a subpopulation of cortical astrocytes expresses GFAP [40].…”

Section: Discussionmentioning

confidence: 99%

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Versatile and Simple Approach to Determine Astrocyte Territories in Mouse Neocortex and Hippocampus

Grosche

Tackenberg

et al. 2013

PLoS ONE

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BackgroundBesides their neuronal support functions, astrocytes are active partners in neuronal information processing. The typical territorial structure of astrocytes (the volume of neuropil occupied by a single astrocyte) is pivotal for many aspects of glia–neuron interactions.MethodsIndividual astrocyte territorial volumes are measured by Golgi impregnation, and astrocyte densities are determined by S100β immunolabeling. These data are compared with results from conventionally applied methods such as dye filling and determination of the density of astrocyte networks by biocytin loading. Finally, we implemented our new approach to investigate age-related changes in astrocyte territories in the cortex and hippocampus of 5- and 21-month-old mice.ResultsThe data obtained by our simplified approach based on Golgi impregnation were compared to previously published dye filling experiments, and yielded remarkably comparable results regarding astrocyte territorial volumes. Moreover, we found that almost all coupled astrocytes (as indicated by biocytin loading) were immunopositive for S100β. A first application of this new experimental approach gives insight in age-dependent changes in astrocyte territorial volumes. They increased with age, while cell densities remained stable. In 5-month-old mice, the overlap factor was close to 1, revealing little or no interdigitation of astrocyte territories. However, in 21-month-old mice, the overlap factor was more than 2, suggesting that processes of adjacent astrocytes interdigitate.ConclusionHere we verified the usability of a simple, versatile method for assessing astrocyte territories and the overlap factor between adjacent territories. Second, we found that there is an age-related increase in territorial volumes of astrocytes that leads to loss of the strict organization in non-overlapping territories. Future studies should elucidate the physiological relevance of this adaptive reaction of astrocytes in the aging brain and the methods presented in this study might be a powerful tool to do so.

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“…For example, Matthias and colleagues reported that within the hippocampus subsets of GFAP expressing cells expressed either glutamate transporters or glutamate receptors (23). Moreover, astrocytes throughout the brain differentially express connexins (24) and GABA and glutamate receptors (26) and different astrocyte populations are reported to differentially support developmental functions and synapse formation (28, 29). Thus, our understanding of the functions of astrocytes is advancing, but much is yet to be learned.…”

Section: Classification Of Non-neuronal Cells In the Brainmentioning

confidence: 99%

Non-Neuronal Cells in the Hypothalamic Adaptation to Metabolic Signals

et al. 2017

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Although the brain is composed of numerous cell types, neurons have received the vast majority of attention in the attempt to understand how this organ functions. Neurons are indeed fundamental but, in order for them to function correctly, they rely on the surrounding “non-neuronal” cells. These different cell types, which include glia, epithelial cells, pericytes, and endothelia, supply essential substances to neurons, in addition to protecting them from dangerous substances and situations. Moreover, it is now clear that non-neuronal cells can also actively participate in determining neuronal signaling outcomes. Due to the increasing problem of obesity in industrialized countries, investigation of the central control of energy balance has greatly increased in attempts to identify new therapeutic targets. This has led to interesting advances in our understanding of how appetite and systemic metabolism are modulated by non-neuronal cells. For example, not only are nutrients and hormones transported into the brain by non-neuronal cells, but these cells can also metabolize these metabolic factors, thus modifying the signals reaching the neurons. The hypothalamus is the main integrating center of incoming metabolic and hormonal signals and interprets this information in order to control appetite and systemic metabolism. Hence, the factors transported and released from surrounding non-neuronal cells will undoubtedly influence metabolic homeostasis. This review focuses on what is known to date regarding the involvement of different cell types in the transport and metabolism of nutrients and hormones in the hypothalamus. The possible involvement of non-neuronal cells, in particular glial cells, in physiopathological outcomes of poor dietary habits and excess weight gain are also discussed.

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Dual reporter approaches for identification of Cre efficacy and astrocyte heterogeneity

Cited by 30 publications

References 41 publications

Astrocytic calcium release mediates peri-infarct depolarizations in a rodent stroke model

Astrocytic calcium release mediates peri-infarct depolarizations in a rodent stroke model

Versatile and Simple Approach to Determine Astrocyte Territories in Mouse Neocortex and Hippocampus

Non-Neuronal Cells in the Hypothalamic Adaptation to Metabolic Signals

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