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GFAP promoter-controlled EGFP-expressing transgenic mice: a tool to visualize astrocytes and astrogliosis in living brain tissue.
- Source :
-
Glia [Glia] 2001 Jan; Vol. 33 (1), pp. 72-86. - Publication Year :
- 2001
-
Abstract
- We have generated transgenic mice in which astrocytes are labeled by the enhanced green fluorescent protein (EGFP) under the control of the human glial fibrillary acidic protein (GFAP) promoter. In all regions of the CNS, such as cortex, cerebellum, striatum, corpus callosum, hippocampus, retina, and spinal cord, EGFP-positive cells with morphological properties of astrocytes could be readily visualized by direct fluorescence microscopy in living brain slices or whole mounts. Also in the PNS, nonmyelinating Schwann cells from the sciatic nerve could be identified by their bright green fluorescence. Highest EGFP expression was found in the cerebellum. Already in acutely prepared whole brain, the cerebellum appeared green-yellowish under normal daylight. Colabeling with GFAP antibodies revealed an overlap with EGFP in the majority of cells. Some brain areas, however, such as retina or hypothalamus, showed only low levels of EGFP expression, although the astrocytes were rich in GFAP. In contrast, some areas that were poor in immunoreactive GFAP were conspicuous for their EGFP expression. Applying the patch clamp technique in brain slices, EGFP-positive cells exhibited two types of membrane properties, a passive membrane conductance as described for astrocytes and voltage-gated channels as described for glial precursor cells. Electron microscopical investigation of ultrastructural properties revealed EGFP-positive cells enwrapping synapses by their fine membrane processes. EGFP-positive cells were negative for oligodendrocyte (MAG) and neuronal markers (NeuN). As response to injury, i.e., by cortical stab wounds, enhanced levels of EGFP expression delineated the lesion site and could thus be used as a live marker for pathology.
- Subjects :
- Animals
Brain metabolism
Brain ultrastructure
Gene Expression Regulation physiology
Glial Fibrillary Acidic Protein metabolism
Gliosis genetics
Gliosis pathology
Green Fluorescent Proteins
Immunohistochemistry
Mice
Mice, Transgenic anatomy & histology
Microscopy, Electron
Neurons cytology
Neurons metabolism
Oligodendroglia cytology
Oligodendroglia metabolism
Peripheral Nerves metabolism
Peripheral Nerves ultrastructure
Astrocytes metabolism
Astrocytes ultrastructure
Glial Fibrillary Acidic Protein genetics
Luminescent Proteins analysis
Luminescent Proteins genetics
Mice, Transgenic genetics
Promoter Regions, Genetic physiology
Subjects
Details
- Language :
- English
- ISSN :
- 0894-1491
- Volume :
- 33
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Glia
- Publication Type :
- Academic Journal
- Accession number :
- 11169793