GFAP promoter-controlled EGFP-expressing transgenic mice: A tool to visualize astrocytes and astrogliosis in living brain tissue

Nolte, Christiané; Matyash, Marina; Pivneva, T. A.; Schipke, Carola G.; Ohlemeyer, Carsten; Hanisch, Uwe‐Karsten; Kirchhoff, Frank; Kettenmann, Helmut

doi:10.1002/1098-1136(20010101)33:1<72::aid-glia1007>3.0.co;2-a

Cited by 504 publications

(402 citation statements)

References 31 publications

Supporting

Mentioning

378

Contrasting

Unclassified

Order By: Relevance

“…Astrocytes were isolated from the forebrains of neonatal (P2) CD1 mice and transgenic mice expressing eGFP under the control of human GFAP promoter (Nolte et al, 2001) or in knockin constructs within the CX3CR1 gene (Jung et al, 2000). Meninges were removed and the tissue was minced with razor blades.…”

Section: Astroglial and Microglia-enriched Culturesmentioning

confidence: 99%

See 1 more Smart Citation

Role of CX3CR1 (Fractalkine Receptor) in Brain Damage and Inflammation Induced by Focal Cerebral Ischemia in Mouse

Dénes

Ferenczi

Halász

et al. 2008

J Cereb Blood Flow Metab

252

220

View full text Add to dashboard Cite

CX3CR1 (fractalkine receptor) is important for sustaining normal microglial activity in the brain. Lack of CX3CR1 reportedly results in neurotoxic microglial phenotype in disease models. The objective of this study was to test the hypothesis that the absence of CX3CR1 worsens the outcome in cerebral ischemia. We observed significantly smaller (56%) infarcts and blood-brain barrier damage in CX3CR1-deficient (CX3CR1À/À) animals compared with CX3CR1 + /À and wild-type mice after transient occlusion of the middle cerebral artery (MCAo). Functional recovery of CX3CR1À/À animals was enhanced, while less number of apoptotic cells and infiltrating leukocytes were found in the ipsilateral hemisphere. Expression of IL-1b mRNA, protein, and interleukin (IL)-1Ra and tumor necrosis factor (TNF)-a mRNAs was lower in CX3CR1À/À mice, whereas no difference was observed in the number of IL-1b-expressing microglia or plasma IL-1b concentration. We observed early IL-1b expression in astrocytes in vivo after MCAo and after oxygen-glucose deprivation in vitro, which might contribute to the ischemic damage. Our findings indicate that lack of CX3CR1 does not result in microglial neurotoxicity after MCAo, but rather significantly reduces ischemic damage and inflammation. Reduced IL-1b and TNFa expression as well as decreased leukocyte infiltration might be involved in the development of smaller infarcts in CX3CR1À/À animals.

show abstract

Section: Astroglial and Microglia-enriched Culturesmentioning

confidence: 99%

“…The contaminating microglial (GFP-positive) cells were removed from the cultures by fluorescent cell sorting. In the second set of experiments, cultures were prepared from the forebrains of hGFAP-GFP mice (Nolte et al, 2001). Astrocytes expressing GFP were subjected to FACS before the experiments.…”

Section: Fluorescence-activated Cell Sortingmentioning

confidence: 99%

Role of CX3CR1 (Fractalkine Receptor) in Brain Damage and Inflammation Induced by Focal Cerebral Ischemia in Mouse

Dénes

Ferenczi

Halász

et al. 2008

J Cereb Blood Flow Metab

252

220

View full text Add to dashboard Cite

show abstract

“…The Otc spf mouse has been widely used as a viable model for the analysis of HAE in UCDs. To define molecular changes that specifically occur in astrocytes during HAE, we crossed the Otc spf mouse with the hGFAPEGFP mouse, in which enhanced green fluorescent protein (EGFP) is selectively expressed in astrocytes (Nolte et al, 2001;Wehner et al, 2003). This enabled us to acutely purify astrocytes from hyperammonemic mouse brains by Fluorescence Activated Cell Sorting (FACS).…”

Section: Introductionmentioning

confidence: 99%

Gene expression profiling of astrocytes from hyperammonemic mice reveals altered pathways for water and potassium homeostasis in vivo

Lichter‐Konecki

Mangin

Gordish‐Dressman

et al. 2008

Glia

View full text Add to dashboard Cite

Acute hyperammonemia (HA) causes cerebral edema and brain damage in children with urea cycle disorders (UCDs) and in patients in acute liver failure. Chronic HA is associated with developmental delay and mental retardation in children with UCDs, and with neuropsychiatric symptoms in patients with chronic liver failure. Astrocytes are a major cellular target of hyperammonemic encephalopathy, and changes occurring in these cells are thought to be causally related to the brain edema of acute HA. To study the effect of HA on astrocytes in vivo, we crossed the Otc spf mouse, a mouse with the X-linked UCD ornithine transcarbamylase (OTC) deficiency, with the hGFAP-EGFP mouse, a mouse selectively expressing green fluorescent protein in astrocytes. We used FACS to purify astrocytes from the brains of hyperammonemic and healthy Otc spf /GFAP-EGFP mice. RNA isolated from these astrocytes was used in microarray expression analyses and qRT-PCR. When compared with healthy littermates, we observed a significant downregulation of the gap-junction channel connexin 43 (Cx43) the water channel aquaporin 4 (Aqp4) genes, and the astrocytic inward-rectifying potassium channel (Kir) genes Kir4.1 and Kir5.1 in hyperammonemic mice. Aqp4, Cx43, and Kir4.1/ Kir5.1 are co-localized to astrocytic end-feet at the brain vasculature, where they regulate potassium and water transport.Since, ions can permeate water and K + -channels, downregulation of these three channels may be a direct effect of elevated blood ammonia levels. Our results suggest that alterations in astrocyte-mediated water and potassium homeostasis in brain may be key to the development of the brain edema.

show abstract

“…It was often the case that weakly fluorescent cells were apparent in the nerves, in particular at the end of the period of OGD. These cells presumably corresponded to the population of dull cells reported previously (Nolte et al, 2001), and appeared to have a high resistance to OGD and were not included in this study.…”

Section: Ischemic Injury To Astrocytes In the Optic Nervementioning

confidence: 99%

“…However, it must be born in mind that GFAP-GFP( + ) of the neonatal subventricular zone have many of the properties of mature astrocytes and behave functionally as astrocytes (Liu et al, 2006), while it is known that GFAP( + ) cells of the neonatal rodent optic nerve are postmitotic in situ (Skoff, 1990). Furthermore, GFAP-GFP expression in somewhat more mature optic nerve is restricted to astrocytes identified on ultrastructural, electrophysiologic, morphologic, and immunolabeling criteria (Nolte et al, 2001). Consistent with this, Steiner et al, 2004 have reported that neurogenic hippocampal astrocytes become GFAP-GFP(À) before expression of the earliest neuronal markers (Steiner et al, 2004).…”

Section: Identity Of Gfap-gfp( + ) Cells In the Neonatal Optic Nervementioning

confidence: 99%

The Mechanisms of Acute Ischemic Injury in the Cell Processes of Developing White Matter Astrocytes

Salter

Fern

2007

J Cereb Blood Flow Metab

View full text Add to dashboard Cite

Astrocytes are fundamentally important to the maintenance and proper functioning of the central nervous system. During the period of development when myelination is occurring, white matter astrocytes are particularly sensitive to ischemic injury and their failure to regulate glutamate during ischemic conditions may be an important factor in excitotoxic injury. Here, we have identified key mechanisms of injury that operate on the processes of immature white matter astrocytes during oxygen-glucose deprivation (OGD) using GFAP-GFP mice. Oxygen-glucose deprivation produced a parallel loss of astrocyte processes and somata, assessed by both the retention of GFP fluorescence within these structures and by quantitative electron microscopy. Oxygen-glucose deprivation-induced process loss was Ca 2 + independent and had two distinct mechanisms. Substituting either extracellular Na + or Cl À , or perfusion with the Na-K-Cl co-transport blocker bumetanide, provided protection up to 40 mins of OGD but not beyond that point. HCO 3 À substitution or perfusion with 4,4 0 -diisothiocyanostilbene-2,2 0 -disulphonic acid provided complete protection of the processes up to 60 mins of OGD. Zero-Na + /zero-K + conditions provided complete protection from OGD-induced injury of processes and somata at all time points. We conclude that acute ischemic-type injury of immature astrocytes follows a cytotoxic ion influx mediated in part by Na-K-Cl co-transport and in part by Na + -and K + -dependent HCO 3 À transport, a mechanism that is common to both cell processes and somata. This work provides a basis on which preventative strategies may be developed to protect white matter astrocytes from ischemic injury in susceptible individuals.

show abstract

GFAP promoter-controlled EGFP-expressing transgenic mice: A tool to visualize astrocytes and astrogliosis in living brain tissue

Cited by 504 publications

References 31 publications

Role of CX3CR1 (Fractalkine Receptor) in Brain Damage and Inflammation Induced by Focal Cerebral Ischemia in Mouse

Role of CX3CR1 (Fractalkine Receptor) in Brain Damage and Inflammation Induced by Focal Cerebral Ischemia in Mouse

Gene expression profiling of astrocytes from hyperammonemic mice reveals altered pathways for water and potassium homeostasis in vivo

The Mechanisms of Acute Ischemic Injury in the Cell Processes of Developing White Matter Astrocytes

Contact Info

Product

Resources

About