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Figure 1


Fig. 1. Construction and PC-specific expression of the L7-pepE transgene. (A) The L7-pepE transgene was made by insertion of a synthetic mini-gene coding for the highly conserved 25 C-terminal amino acids of synapsin Ia (pepE; see box) into the L7{Delta}AUG vector. (B) PCR analysis of DNA prepared from tail biopsies revealed insertion of the transgene in four distinct transgenic (Tg-1, Tg-2, Tg-3, Tg-4) mouse lines. WT, wild-type. (C) The expression of the L7-pepE transgene was assessed by RT-PCR experiments performed on total RNA prepared from the cerebella of wild-type (wt) and two L7-pepE (Tg-1 and Tg-2) mouse lines. Samples that were not incubated with reverse transcriptase (RT) are shown as negative controls. HPRT was used as an internal control. (D) In situ hybridization was performed on sagittal (a,b) or coronal (c-f) sections from the brains of wild-type (a,c,e) and Tg-1 (b) or Tg-2 (d,f) transgenic mice using a 35S-labelled L7-pepE antisense oligonucleotide probe. The hybridization signal, undetectable in wild-type sections, is intense in transgenic sections and exclusively restricted to the cerebellar cortex. At higher magnification (e,f), grains counterstained with cresyl violet are concentrated in PC somata. Bars, 2.8 mm (a,b); 1.2 mm (c,d); 100 µm (e,f).





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