High level Purkinje cell specific expression of green fluorescent protein in transgenic mice.
The green fluorescent protein (GFP) has become a powerful tool in molecular and cell biology. It is a commonly used marker for cloning and transfection experiments as well as a useful label of living cells allowing continuous observation of developing structures. In order to unravel mechanisms of neuronal differentiation, we generated a transgenic mouse model which expresses GFPS65T,hu under the control of the Purkinje cell-specific promoter L7/pcp-2. Here, we show that GFPS65T,hu is highly expressed specifically in the cerebellum in whole mount preparations after the 2nd postnatal week. GFPS65T,hu can be detected exclusively in Purkinje cells of cerebellar slices. The fluorescence intensity of GFPS65T,hu should enable the characterization and recording of axons, dendrites, and spines protruding from these neuronal processes. The level of GFP expression can be quantified by western blotting which allows to analyze protein expression and L7/pcp-2 promoter regulation in vivo. The application of cellular and physiological techniques on L7GFP mice will provide a remarkable opportunity to investigate various aspects of neuronal development at the cellular and subcellular levels.
Zhang
X
X
Department of Neuroscience, Neurobiotechnology Center, The Ohio State University, Columbus, Ohio, USA.
Baader
S L
SL
Bian
F
F
Müller
W
W
Oberdick
J
J
D000818:Animals; D002531:Cerebellum / Q000378:metabolism / Q000473:pathology; D005260:Female; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011689:Purkinje Cells / Q000378:metabolism*; D013347:Subcellular Fractions
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins