Cytochemical analysis of storage materials in cultured skin fibroblasts from patients with I-cell disease.
BACKGROUND
In cultured fibroblasts from I-cell disease patients the transport of many lysosomal enzymes is defective, and affected cells contain inclusion bodies filled with undegraded substrates. However, the contents of these inclusion bodies have not been well characterized yet. We attempted to identify accumulated substances in cultured I-cell disease fibroblasts cytochemically.


METHODS
Cultured fibroblasts from I-cell disease patients were double-stained with a monoclonal antibody to lysosome-associated membrane protein-1 (LAMP-1) and that to GM2 ganglioside, or a series of lectins that specifically bind to sugar moieties.


RESULTS
The patients' cells were granularly stained with the antibody to GM2 ganglioside and the lectins including Maakia amurensis, Datura stramonium, and concanavalin A. Their localization was coincident with that of LAMP-1.


CONCLUSIONS
GM2 ganglioside and various kinds of glycoconjugates having sialic acidalpha2-3galactose, galactosebeta1-4N-acetylglucosamine and mannose residues accumulate in enlarged lysosomes in I-cell disease fibroblasts.
Kawashima
Ikuo
I
Department of Clinical Genetics, The Tokyo Metropolitan Institute of Medical Science, Tokyo Metropolitan Organization for Medical Research, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan.
Ohsawa
Mai
M
Fukushige
Tomoko
T
Nagayama
Yoshihisa
Y
Niida
Yo
Y
Kotani
Masaharu
M
Tajima
Youichi
Y
Kanekura
Takuro
T
Kanzaki
Tamotsu
T
Sakuraba
Hitoshi
H
D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000737:chemistry*; D005678:G(M2) Ganglioside / Q000032:analysis*; D006651:Histocytochemistry; D006801:Humans; D007223:Infant; D037102:Lectins / Q000032:analysis; D051907:Lysosomal Membrane Proteins / Q000032:analysis*; D008247:Lysosomes / Q000201:enzymology; D008854:Microscopy, Electron; D009081:Mucolipidoses / Q000378:metabolism* / Q000473:pathology; D012867:Skin / Q000648:ultrastructure; D052118:Lysosomal-Associated Membrane Protein 1
D005678:G(M2) Ganglioside; D037102:Lectins; D051907:Lysosomal Membrane Proteins; C489116:LAMP1 protein, human; D052118:Lysosomal-Associated Membrane Protein 1