GFAP promoter-controlled EGFP-expressing transgenic mice: a tool to visualize astrocytes and astrogliosis in living brain tissue.
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.
Nolte
C
C
Max Delbrück Center for Molecular Medicine, Cellular Neurosciences, Berlin-Buch, Germany.
Matyash
M
M
Pivneva
T
T
Schipke
C G
CG
Ohlemeyer
C
C
Hanisch
U K
UK
Kirchhoff
F
F
Kettenmann
H
H
D000818:Animals; D001253:Astrocytes / Q000378:metabolism* / Q000648:ultrastructure*; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure; D005786:Gene Expression Regulation / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics* / Q000378:metabolism; D005911:Gliosis / Q000235:genetics / Q000473:pathology; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic / Q000033:anatomy & histology / Q000235:genetics*; D008854:Microscopy, Electron; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism; D010525:Peripheral Nerves / Q000378:metabolism / Q000648:ultrastructure; D011401:Promoter Regions, Genetic / Q000502:physiology*
D005904:Glial Fibrillary Acidic Protein; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins