Live astrocytes visualized by green fluorescent protein in transgenic mice.
Green fluorescent protein (hGFP-S65T) was expressed in transgenic mice under the control of the astrocyte-specific glial fibrillary acidic protein (GFAP) promoter. Tissues from two independent transgenic lines were characterized by Northern blot analysis and by confocal microscopy. The expression pattern in these two lines was identical in all tissues examined, and similar to that found previously with a lacZ transgene driven by the same promoter. Bright fluorescence was observed in the cell bodies and processes of unfixed or fixed astrocytes, using both whole mount and brain slice preparations, from multiple areas of the central nervous system. However, in contrast to GFAP-lacZ transgenics, retinal Müller cells expressed the GFP transgene in response to degeneration of neighboring photoreceptors. These data indicate that the 2.2-kb hGFAP promoter contains sufficient regulatory elements to direct expression in Müller cells, and that GFP is a suitable reporter gene for use in living preparations of the mammalian nervous system. Such mice should prove useful for studies of dynamic changes in astrocyte morphology during development, and in response to physiological and pathological conditions.
Zhuo
L
L
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 53706, USA.
Sun
B
B
Zhang
C L
CL
Fine
A
A
Chiu
S Y
SY
Messing
A
A
D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D001921:Brain / Q000166:cytology / Q000502:physiology; D017930:Genes, Reporter; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis / Q000235:genetics; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000096:biosynthesis*; D051379:Mice; D008822:Mice, Transgenic; D009900:Optic Nerve / Q000502:physiology; D010786:Photoreceptor Cells / Q000166:cytology / Q000502:physiology; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012160:Retina / Q000166:cytology / Q000502:physiology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D005904:Glial Fibrillary Acidic Protein; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase