ABSTRACT
Brain tissue was obtained promptly after death from a patient with autosomal dominant olivopontocerebellar atrophy and studied by immunocytochemistry and a Golgi technique. Antiglutamic acid decarboxylase showed severe loss of Purkinje cells and their terminals in the dentate nucleus. Stains for neuron-specific enolase (NSE) and microtubule-associated proteins (MAP) confirmed the integrity of the dentate nucleus. Basket and stellate cells revealed secondary changes, but Golgi neurons were intact. Methods for NSE and MAP disclosed dendritic alterations and loss of neurons in the basis pontis and inferior olivary nuclei. Golgi impregnation of Purkinje cells showed loss of major dendrites, paucity of spiny branchlets, and axonal expansions.
Subject(s)
Olivopontocerebellar Atrophies/pathology , Spinocerebellar Degenerations/pathology , Aged , Cerebellum/ultrastructure , Cerebral Cortex/pathology , Cerebral Cortex/ultrastructure , Glutamate Decarboxylase , Humans , Immunochemistry , Male , Microtubule-Associated Proteins , Middle Aged , Olivopontocerebellar Atrophies/metabolism , Olivopontocerebellar Atrophies/physiopathology , Phosphopyruvate Hydratase , Purkinje Cells/ultrastructureABSTRACT
Amyloid fibrils were isolated from the myocardium of two patients with familial amyloid polyneuropathy. The solubilized amyloid fibril whole protein shared immunologic determinants with normal human serum prealbumin (transthyretin), but revealed subtle differences on immunoelectrophoresis and radial immunodiffusion. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, amyloid fibril whole protein was resolved into numerous bands that reacted with antitransthyretin on immunoblots. The whole protein also contained peptide fragments of fibronectin, but was devoid of amyloid P protein. An antiserum raised against the whole protein was suitable for immunocytochemistry of amyloid in paraffin sections. In contrast, commercial antitransthyretin, raised against the intact tetrameric protein failed to react with tissue amyloid. Immunochemical and immunocytochemical results support the concept that familial amyloid polyneuropathy with cardiomyopathy is due to infiltration of susceptible tissues by an anomalous transthyretin.
Subject(s)
Amyloidosis/genetics , Nervous System Diseases/genetics , Aged , Amyloid/isolation & purification , Amyloidosis/metabolism , Amyloidosis/pathology , Female , Histocytochemistry , Humans , Immunochemistry , Male , Microscopy, Electron , Middle Aged , Myocardium/analysis , Nervous System/analysis , Nervous System Diseases/metabolism , Nervous System Diseases/pathology , Pedigree , Prealbumin/immunologyABSTRACT
Free malonate, malonyl-coenzyme A (malonyl-CoA), and acetyl-CoA were assayed in rat brain at developmental ages from the 20th day of gestation to 60 days of postnatal life. The determination of malonate was based on its conversion to malonyl-CoA and decarboxylation to acetyl-CoA by enzyme extracts from Pseudomonas fluorescens. The resulting acetyl-CoA reacted with [4-14C]oxaloacetate to form [5-14C]citrate, which was isolated by TLC. Malonyl-CoA in perchloric acid extracts from brain was converted to acetyl-CoA by rat liver mitochondrial malonyl-CoA decarboxylase (EC 4.1.1.9). Acetyl-CoA derived from this step was assayed by a modified CoA-cycling procedure. Brain acetyl-CoA was also assayed by CoA cycling. Prenatal brain contained no free malonate but malonyl-CoA was present. The acetyl-CoA level was relatively high just prior to birth and declined slightly with growth. Malonate concentrations after birth rose rapidly to reach 192 nmol/g wet weight at 60 days. Adult levels for malonyl-CoA and acetyl-CoA were 1.83 and 1.90 nmol/g wet weight, respectively. The origin and natural role of free malonate in brain are not known but deacylation of malonyl-CoA by reversal of the malonyl-CoA synthetase reaction is postulated. Rat liver and kidney also contain substantial concentrations of free malonate.
Subject(s)
Acetyl Coenzyme A/analysis , Acyl Coenzyme A/analysis , Bacterial Proteins , Brain/growth & development , Malonates/analysis , Malonyl Coenzyme A/analysis , Aging , Animals , Brain/embryology , Carboxy-Lyases/metabolism , Coenzyme A Ligases/metabolism , Female , Gestational Age , Pregnancy , Pseudomonas fluorescens/enzymology , Rats , Rats, Inbred StrainsABSTRACT
In-vitro fatty acid biosynthesis was studied in normal rat sciatic nerve and during wallerian degeneration. Normal nerve incorporated 1,3-C14-malonyl-CoA and 1-C14-acetyl-CoA into fatty acids by a de-novo biosynthetic pathway. The reaction product with highest radioactivity was palmitic acid, and the free fatty acids of nerve contained 90% of the total fatty acid label. During wallerian degeneration, there was a rapid increase of fatty acid biosynthesis that reached a peak between 16 and 24 days after nerve section. Values declined to normal levels at approximately 50 days.
Subject(s)
Fatty Acids/biosynthesis , Nerve Degeneration , Sciatic Nerve/metabolism , Wallerian Degeneration , Acetyl Coenzyme A/metabolism , Amino Acids/metabolism , Animals , Carbon Radioisotopes , Hydrogen-Ion Concentration , Lipids/biosynthesis , Malonyl Coenzyme A/metabolism , Proteins/metabolism , RatsSubject(s)
Brain/metabolism , Carboxy-Lyases/metabolism , Malonates/metabolism , Mitochondria/metabolism , Sulfurtransferases/metabolism , Thiolester Hydrolases/metabolism , Acetyl Coenzyme A/metabolism , Acetyl-CoA Carboxylase/metabolism , Animals , Brain/cytology , Brain/enzymology , Carbon Dioxide/metabolism , Carbon Radioisotopes , Coenzyme A , Coenzyme A Ligases/metabolism , Keto Acids , Liver/enzymology , Male , Mitochondria/enzymology , Rats , Succinates , Surface-Active Agents , UltrasonicsABSTRACT
Following ultracentrifugation in sucrose and hypotonic exposure ('osmotic shock'), myelin fractions were prepared from homogenates of human centrum ovale obtained post-mortem. Non-specific esterase (NsE), succinic dehydrogenase and acid phosphatase activities were determined. One isolation procedure utilized aqueous sucrose solutions only. Other preparative technics involved addition of phosphate buffer (pH 6.6 or 8.0) or varying concentrations of calcium or magnesium ions to preparative media. The NsE activity of myelin increased greatly when electrolytes were included in these preparative solutions. Morphologic homogeneity and protein and lipid contents of myelin were similar whatever the isolation procedure.
