Prevention of lysosomal storage in Tay-Sachs mice treated with N- butyldeoxynojirimycin

Science

Volumn 276, Issue 5311, 1997, Pages 428-431

  Platt, Frances M ^a   Neises, Gabrielle R ^b   Reinkensmeier, Gabriele ^a   Townsend, Mandy J ^c   Perry, V Hugh ^c   Proia, Richard L ^d   Winchester, Bryan ^e   Dwek, Raymond A ^a   Butters, Terry D ^a  

^a UNIVERSITY OF OXFORD   (United Kingdom)

^b MONSANTO COMPANY   (United States)

^c UNIVERSITY OF OXFORD   (United Kingdom)

^d NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES   (United States)

^e UNIVERSITY COLLEGE LONDON   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

BETA N ACETYLHEXOSAMINIDASE; GANGLIOSIDE GM2; GLUCOSYLCERAMIDE; GLYCOSPHINGOLIPID; MIGLUSTAT;

ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; BLOOD BRAIN BARRIER; CONTROLLED STUDY; DRUG BLOOD LEVEL; ENZYME SUBSTRATE; LYSOSOME STORAGE DISEASE; MOUSE; NONHUMAN; PRIORITY JOURNAL; TAY SACHS DISEASE;

ANIMALIA;

EID: 0030937840     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.276.5311.428     Document Type: Article

References (20)

1. E. F. Neufeld, Annu. Rev. Biochem. 60, 257 (1991).
2. E. Beutler, Science 256, 794 (1992).
3. N. W. Barton et al., N. Engl. J. Med. 324, 1464 (1991).
4. E. Beutler et al., Blood 78, 1183 (1991).
5. F. M. Platt, G. R. Neises, R. A. Dwek, T. D. Butters, J. Biol. Chem. 269, 8362 (1994); F. M. Platt, G. R. Neises, G. B. Kartsson, R. A. Dwek, T. D. Butters, ibid., p. 27108.
6. F. M. Platt, G. R. Neises, R. A. Dwek, T. D. Butters, J. Biol. Chem. 269, 8362 (1994); F. M. Platt, G. R. Neises, G. B. Kartsson, R. A. Dwek, T. D. Butters, ibid., p. 27108.
7. F. M. Platt and T. D. Butters, Trends Glycosci. Glycotechnol. 7, 495 (1995).
8. F. M. Platt, G. Reinkensmeier, R. A. Dwek, T. D. Butters, in preparation.
9. S. Yamanaka et al., Proc. Natl. Acad. Sci. U.S.A. 91, 9975 (1994).
10. K. Sandhoff, E. Conzelmann, E. F. Neufeld, M. M. Kaback, K. Suzuki, in The Metabolic Basis of Inherited Disease, C. R. Scriver, A. L. Beaudet, W. S. Sly, D. Valle, Eds. (McGraw-Hill, New York, 1989), vol. 2, pp. 1807-1839.
11. K. Sango et al., Nature Genet. 11, 170 (1995).
12. M. Taniike et al., Acta Neuropathol. 89, 296 (1995).
13. From weaning (4 weeks), mice were fed a diet of powdered mouse chow (expanded Rat and Mouse Chow 1, ground, SDS Ltd., Witham, Essex, UK) containing NB-DNJ. The diet and compound (both dry solids) were mixed thoroughly before use, stored at room temperature, and used within 7 days of mixing. Water was available to the mice ad libitum. The mice were housed under standard nonsterite conditions and were given 4800 mg per kilogram of body weight per day of NB-DNJ, which gave serum concentrations of ∼50 μM (data not shown).
14. M. A. Fischl et al., J. Acquired Immune Defic. Syndr. 7, 139 (1994).
15. The animals were anesthetized, perfused with phosphate-buffered saline (PBS) (pH 7.2), and the intact brain removed. The brain tissue was manually homogenized in water, freeze-dried, and extracted twice with chloroform:methanol 2:1 (v/v) for 2 hours at room temperature and overnight at 4°C. A volume of the solvent extract equivalent to 5 mg dry weight for each brain was base hydrolyzed (70), and the Folch upper phase was separated by TLC (Silica gel 60 plates, Merck, British drug house, Poole, Dorset, UK) in chloroform:methanol: 0.22% calcium chloride (60:35:8), then sprayed with orcinol and visualized by heating to 80°C for 10 min.
16. Mice were anesthetized, perfused with PBS (pH 7.4) containing 4% paraformaldehyde, and the brain dissected and retained in fixative overnight prior to cryopreservation and sectioning. Frozen brain sections (7 μm) were warmed to room temperature, stained with PAS according to the manufacturer's instructions (Sigma, Poole, Dorset, UK), counterstained with Erhlich's hematoxylin, and mounted in diethyl-(phenyl)xanthine (British drug house).
17. The mice were anesthetized and perfusion fixed with 2% paraformaldehyde, 2% glutaraldehyde mix in PBS. The brain was dissected and fixed in the same fixative overnight at 4°C. The brain was trimmed, and 100-μm sections were cut on a vibrotome, then washed three times in 0.1 M phosphate buffer and stained with osmium tetroxide (1% in 0.1 M phosphate) for 35 min. The sections were dehydrated through an ethanol series, treated with propylene oxide (twice for 15 min), and then placed in Durcupan resin overnight at room temperature, transferred to glass slides, and kept at 60°C for 48 hours. Storage areas of the brain were selected microscopically, cut out of the thick section with a scalpel blade, and transferred to propylene oxide and embedded in Embed 800 (Electron Microscopy Sciences, Fort Washington, PA). Sections were stained with uranyl acetate/lead citrate and observed with a Hitachi 600 microscope at 75 kV.
18. T. Butters, unpublished observation.
19. F. Platt, unpublished observation.
20. We thank Searle/Monsanto for NB-DNJ, J. Vass and D. Smith for excellent technical assistance, and C. Beesley for photography. The Glycobiology Institute is supported by Searle/Monsanto. F.M.P. is a Lister Institute Research Fellow.