title: The murine stem cell virus promoter drives correlated transgene expression in the leukocytes and cerebellar Purkinje cells of transgenic mice.
abstract: The murine stem cell virus (MSCV) promoter exhibits activity in mouse hematopoietic cells and embryonic stem cells. We generated transgenic mice that expressed enhanced green fluorescent protein (GFP) under the control of the MSCV promoter. We obtained 12 transgenic founder mice through 2 independent experiments and found that the bodies of 9 of the founder neonates emitted different levels of GFP fluorescence. Flow cytometric analysis of circulating leukocytes revealed that the frequency of GFP-labeled leukocytes among white blood cells ranged from 1.6% to 47.5% across the 12 transgenic mice. The bodies of 9 founder transgenic mice showed various levels of GFP expression. GFP fluorescence was consistently observed in the cerebellum, with faint or almost no fluorescence in other brain regions. In the cerebellum, 10 founders exhibited GFP expression in Purkinje cells at frequencies of 3% to 76%. Of these, 4 mice showed Purkinje cell-specific expression, while 4 and 2 mice expressed GFP in the Bergmann glia and endothelial cells, respectively. The intensity of the GFP fluorescence in the body was relative to the proportion of GFP-positive leukocytes. Moreover, the frequency of the GFP-expressing leukocytes was significantly correlated with the frequency of GFP-expressing Purkinje cells. These results suggest that the MSCV promoter is useful for preferentially expressing a transgene in Purkinje cells. In addition, the proportion of transduced leukocytes in the peripheral circulation reflects the expression level of the transgene in Purkinje cells, which can be used as a way to monitor transgene expression properties in the cerebellum without invasive techniques.
authors.affiliation: Department of Neurophysiology, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Oue
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Handa
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Matsuzaki
authors.forename: Kazutomo
authors.initials: K
authors.lastname: Suzue
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Murakami
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Hirai
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D018703:Founder Effect; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics; D007962:Leukocytes / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D032342:Muscle Cells / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011689:Purkinje Cells / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics*; D019076:Transgenes / Q000235:genetics*
chemical_list: D049452:Green Fluorescent Proteins