Immunohistochemical characterization of transgenic mice highly expressing human lysosomal alpha-galactosidase

Biochim Biophys Acta. 1998 Apr 28;1406(3):260-6. doi: 10.1016/s0925-4439(98)00012-x.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Northern
  • Blotting, Southern
  • Blotting, Western
  • Female
  • Gene Expression*
  • Humans
  • Immunohistochemistry
  • Kidney / enzymology
  • Liver / enzymology
  • Lysosomes / enzymology*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Myocardium / enzymology
  • Organ Specificity / genetics
  • alpha-Galactosidase / biosynthesis*
  • alpha-Galactosidase / genetics*
  • alpha-Galactosidase / metabolism

Substances

  • alpha-Galactosidase