Isolation, characterization and in vivo analysis of the murine calbindin-D28K upstream regulatory region.
The genomic locus containing the murine calbindin-D28K gene has been isolated and partially characterized. Genomic cloning revealed an exon/intron chromosomal structure very similar to the avian gene previously described. The ability of the calbindin-D28K upstream region to direct cell-specific expression was tested in vivo. Varying lengths of upstream sequence were used to drive expression of lacZ in transgenic mice. Characterization of 23 transgenic mouse lines revealed that even as much as 3.0 kb of upstream sequence was unable to direct expression independently of integration site effects, suggesting the absence of important elements. Despite the small number of expressing transgenic lines and the great variability, there was a tendency of cell specificity of transgene expression exhibited in distinct brain regions. In the cerebellum, Purkinje cell-specific expression was observed with the shortest (1.0 kb) upstream sequence tested. Specificity of transgene expression in Purkinje cells was abolished with longer portions of upstream sequence. The same observation was made for transgene expression in granule cells of the dentate gyrus, while the opposite effect was observed for expression in CA1 hippocampal cells. The absence of any transgenic lines exhibiting appropriate transgene expression in the kidney suggested that the VDREs described previously for the murine calbindin gene are not sufficient to direct kidney expression in vivo. It is concluded that 3.0 kb of calbindin upstream sequence includes the regulatory elements dictating a portion of cell-specificity in the CNS of transgenic mice, albeit lacking regions that allow expression independently of chromosomal effects.
Pavlou
O
O
Department of Genetics and Cell Biology, University of Minnesota, Minneapolis 55455, USA.
Ehlenfeldt
R
R
Horn
S
S
Orr
H T
HT
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D064092:Calbindin 1; D064026:Calbindins; D018891:Dentate Gyrus / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D064030:S100 Calcium Binding Protein G / Q000235:genetics*
C576499:Calb1 protein, mouse; D064092:Calbindin 1; D064026:Calbindins; D064030:S100 Calcium Binding Protein G