Primary culture of neural cells isolated from the cerebellum of newborn and adult mucopolysaccharidosis type IIIA mice.
In order to evaluate the mechanisms leading to neuropathology in Mucopolysaccharidosis type IIIA (MPS-IIIA, Sanfilippo syndrome), we have harvested and cultured primary neural cells isolated from the cerebellum of newborn and adult MPS-IIIA and unaffected mice. Cell viability and plating efficiency were comparable for brain tissue obtained from either newborn or adult MPS-IIIA and unaffected mice. Cultures (newborn and adult) comprised a mixed brain cell population including astrocytes, oligodendrocytes, and neurons. Newborn MPS-IIIA cells contained inclusions and vacuoles consistent with the pathology present in affected brain tissue. Newborn and adult MPS-IIIA brain cells had approximately 5-7% of the sulfamidase activity present in primary neural cells cultured from unaffected newborn and adult mice. In addition, high levels of glucosamine-N-sulfate[alpha-1,4]hexuronic acid, a heparan sulfate-derived disaccharide, were detected in both newborn and adult MPS-IIIA brain cells. These results suggest that the primary MPS-IIIA brain cells exhibit characteristics of MPS-IIIA phenotype at the histopathological and biochemical level in culture.
Sutherland
L M
LM
Department of Genetic Medicine, Lysosomal Diseases Research Unit, Children, Youth and Women's Health Service, 72 King William Road, North Adelaide, SA 5006, Australia. leanne.sutherland@adelaide.edu.au
Hemsley
K M
KM
Hopwood
J J
JJ
D000367:Age Factors; D000375:Aging / Q000378:metabolism / Q000473:pathology; D000818:Animals; D001253:Astrocytes / Q000378:metabolism / Q000473:pathology; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D002531:Cerebellum / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D004187:Disaccharides / Q000378:metabolism; D004195:Disease Models, Animal; D006603:Hexuronic Acids / Q000378:metabolism; D006867:Hydrolases / Q000378:metabolism; D002479:Inclusion Bodies / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D009084:Mucopolysaccharidosis III / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D009457:Neuroglia / Q000378:metabolism / Q000473:pathology*; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D009836:Oligodendroglia / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D014617:Vacuoles / Q000378:metabolism / Q000473:pathology
D004187:Disaccharides; D006603:Hexuronic Acids; D006867:Hydrolases; C022708:N-sulfoglucosamine sulfohydrolase