title: Immunohistochemical characterization of transgenic mice highly expressing human lysosomal alpha-galactosidase.
abstract: Human lysosomal alpha-galactosidase predominantly hydrolyzes ceramide trihexoside. A transgenic mouse line, C57BL/6CrSIc-TgN(GLA) 1951 Rin, highly expressing human alpha-galactosidase, has been established and investigated biochemically and immunohistochemically in order to clarify the distribution of the expressed enzyme proteins and to evaluate it as a donor model of organ transplantation therapy for Fabry disease caused by a genetic defect of alpha-galactosidase. In these transgenic mice, about five copies of the transgene were integrated, and alpha-galactosidase activity was expressed in liver, kidney, heart, spleen, small intestine, submaxillary gland, skeletal muscle, cerebrum, cerebellum, bone marrow cells and serum. The enzyme activity was about 22 to 11,080-fold higher than that in non-transgenic mice. In liver, heart and kidney tissues, which are important organs for transplantation studies, sufficient amounts of alpha-galactosidase mRNAs were transcribed, and the expressed enzymes, with molecular weights of 54-60 kDa, are abundant in the liver (enzyme activity: 53,965 nmol h-1 mg-1 protein) and heart (39,906 nmol h-1 mg-1 protein), followed by in the kidney tissue (9177 nmol h-1 mg-1 protein), respectively. An immunohistochemical microscopic study clearly demonstrated the distribution of the expressed enzyme proteins in kidney and liver tissues. Highly expressed alpha-galactosidase was detected in glomerular cells, tubular cells and hepatocytes. These transgenic mice will be useful as a donor model for experimental organ transplantation, and also it will enable recurrent biopsies and long-term observation. The organ transplantation data on mice will provide us with important information.
authors.affiliation: Department of Clinical Genetics, the Tokyo Metropolitan Institute of Medical Science, 18-22, Honkomagome-3, Bunkyo-ku, Tokyo 113-8613, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Kase
authors.forename: M
authors.initials: M
authors.lastname: Shimmoto
authors.forename: K
authors.initials: K
authors.lastname: Itoh
authors.forename: K
authors.initials: K
authors.lastname: Utsumi
authors.forename: M
authors.initials: M
authors.lastname: Kotani
authors.forename: C
authors.initials: C
authors.lastname: Taya
authors.forename: H
authors.initials: H
authors.lastname: Yonekawa
authors.forename: H
authors.initials: H
authors.lastname: Sakuraba
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D005260:Female; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity / Q000235:genetics; D000519:alpha-Galactosidase / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism
chemical_list: D000519:alpha-Galactosidase