High level Purkinje cell specific expression of green fluorescent protein in transgenic mice

X Zhang, SL Baader, F Bian, W Müller… - Histochemistry and cell …, 2001 - Springer
X Zhang, SL Baader, F Bian, W Müller, J Oberdick
Histochemistry and cell biology, 2001Springer
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 GFP S65T, hu under the control of the Purkinje cell-specific promoter
L7/pcp-2. Here, we show that GFP S65T, hu is highly expressed specifically in the …
Abstract
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.
Springer
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