ABSTRACT
Two unrelated 9-year-old boys failed to thrive from ages 5 and 4 years, and had focal cerebral seizures followed by transcent hemipareses. Histochemistry of their muscle biopsies showed "ragged-red" fibers, which ultrastructurally contained clusters of mitochondria having loss of crisp delineation of crista membranes and contained amorphous inclusion material and parallel-packed cristae and sometimes paracrystalline inclusions. In the patients' cultured muscles, similar mitochondrial abnormalities were present. 2,4-Dinitrophenol, introduced to the medium of cultures of normal human muscle, produced mitochondrial abnormalities similar to those of the patients', and the medium of the patients' muscle cultures worsened the mitochondrial abnormalities. This study, in demonstrating a mitochondrial defect reproducible in the cultured muscle fibers and, therefore, intrinsic to the ragged-red muscle fibers themselves, raises the possibility of a collateral mitochondrial defect in CNS cells as part of a multicellular mitochondriopathy.
Subject(s)
Acidosis/pathology , Epilepsies, Partial/pathology , Lactates/metabolism , Mitochondria/ultrastructure , Biopsy , Cells, Cultured , Child , Culture Media , DNA , Dinitrophenols/pharmacology , Growth Disorders/pathology , Humans , Inclusion Bodies/ultrastructure , Male , Mitochondria/drug effects , Mitochondria/metabolism , Mitochondria/physiopathology , Muscles/ultrastructureSubject(s)
Gaucher Disease/immunology , Glucosidases/immunology , Glucosylceramidase/immunology , Antibody Formation , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay , Female , Gaucher Disease/drug therapy , Glucosylceramidase/therapeutic use , Humans , Immunoglobulin G/immunology , Immunoglobulin M/immunology , Male , Middle Aged , Placenta/enzymologyABSTRACT
Glucocerebrosidase was purified 26,000-fold from spleens from normal humans and from patients with Gaucher disease (Gaucher spleens). The specific activities of the purified normal and mutant enzymes with glucocerebroside as substrate were 8.5 X 10(5) and 5.4 X 10(4) nmol/mg of protein per hr, respectively. The ratio of enzymatic activities was constant throughout the isolation procedure. The two enzymes appeared to be similar by other parameters such as substrate affinity, heat lability, and pH optimum. Immunotitration with glucocerebrosidase antiserum showed equivalent quantities of crossreacting material in extracts of normal and Gaucher spleens. These data strongly suggest that the genetic basis of Gaucher disease is a strucutral mutation of glucocerebrosidase. The results of sodium dodecyl sulfate gel electrophporesis also indicate that there are differences between the normal and the Gaucher disease enzyme.
