Ceroid Lipofuscinosis Research Articles

Accumulation of phosphorylated dolichol in several tissues in ceroid-lipofuscinosis (Batten disease)

The concentrations of phosphorylated dolichol (P-dolichol) and of non-phos-phorylated dolichol were estimated in a number of different tissues from ceroid-lipofuscinosis (CL) and control cases. The P-dolichol contents of four tissues, brain, pancreas, muscle and kidney were significantly greater than control values. Liver and spleen showed little or no increase in P-dolichol content. Though non-phos-phorylated dolichol levels were higher in CL tissues than in control tissues, the differences were significant only for brain and pancreas. In control muscle, a linear age-related increase in non-phosphorylated dolichol content was observed. The relative amounts of the major homologs of P-dolichol and of non-phosphorylated dolichol were estimated. Each tissue has a characteristic homolog size-distribution, and there is close similarity between the size-distribution profiles of P-dolichol and of non-phosphorylated dolichol for each tissue.

Taurine availability and function in neurogenetic retinopathies.

Several neurogenetic diseases manifest abnormal retinal electrophysiology and pathology; such changes in both retinal integrity and function resemble those found in the retina of individuals with retinitis pigmentosa. Some of these inherited neurodegenerative disease include: ceroid lipofuscinosis, Bardet-Beidl-Lawrence-Moon syndrome and Refsum disease, (36). The etiologies of the retinal degeneration observed in these neurogenetic diseases and retinitis pigmentosa remain unknown. Preliminary results, however, from this laboratory (68) and published reports from other investigators (16,21) suggest altered transport of the amino acid taurine in several neurodegenerative diseases.

Lysosomal storage in human skeletal muscle

Skeletal muscle is involved symptomatically in two lysosomal storage diseases, acid maltase deficiency and a similar condition in which enzyme levels are normal. Asymptomatic storage in skeletal muscle cells is found in Batten-Kufs' disease (ceroid lipofuscinosis), Fabry's disease, and mannosidosis, as well as in rare patients with an unidentified storage disease. Other cell types (vascular endothelium, smooth muscle, fibroblasts, satellite cells) within the confines of the biopsy specimen may reveal storage in other diseases. The differential diagnosis involves predominantly both normal and abnormal conditions in which acid phosphatase activity is prominent in cells.

Ceroid lipofuscinosis in sheep. II. The major component of the lipopigment in liver, kidney, pancreas, and brain is low molecular weight protein.

Previous studies on the lipopigment from the livers of sheep affected with ceroid lipofuscinosis showed that the disease does not involve a defect in lipid metabolism or abnormal lipid peroxidation and that most of the lipopigment was proteinaceous. In this study, lipopigment was isolated from liver, kidney, pancreas, and brain of affected sheep without the use of proteolytic enzymes. Lipopigment from all tissues was two-thirds protein. Modified silver staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a major band of Mr = 14,800, heterogeneous material between Mr = 5,000 and 9,000, and a major band of Mr = 3,500. These compounds did not stain for RNA or carbohydrate and were digested by a nuclease-free protease as expected for protein. They are not normal lysosomal proteins. Lipopigment levels of dolichol, ubiquinone, and cholesterol were consistent with the lipopigment being protein-enriched lysosome-derived cytosomes. The presence of the Mr = 3,500 proteins in whole affected tissue homogenates distinguished them from homogenates of normal tissues. It was concluded that low Mr proteins are specifically stored in ovine ceroid lipofuscinosis and that the ceroid lipofuscinoses may result from inherited defects in lysosomal protein catabolism.

Ceroid lipofuscinosis in sheep. I. Bis(monoacylglycero)phosphate, dolichol, ubiquinone, phospholipids, fatty acids, and fluorescence in liver lipopigment lipids.

The ceroid lipofuscinoses are inherited lysosomal diseases of children characterized by a fluorescent lipopigment stored in a variety of tissues. Defects in lipid metabolism or the control of lipid peroxidation have been postulated to explain their pathogenesis. In the present study, lipopigment was isolated from the liver of sheep affected with ceroid lipofuscinosis. It was 70% protein, the rest being mainly lipids. These were only one-sixth as fluorescent as total liver lipids, but contained a number of fluorophors. None were major components of the lipopigment or the postulated fluorescent product of lipid peroxidation. Lipopigment lipids included the lysosomal marker bis(monoacylglycero)phosphate that contained 42.9% linoleate and 16.5% linolenate. Lipopigment neutral lipids were dolichol, dolichyl esters, ubiquinone, free fatty acids, and cholesterol, indicative of a lysosomal origin of the lipopigment. Phosphatidylcholine, phosphatidylinositol, phosphatidylserine, and phosphatidylethanolamine were present in proportions and with fatty acid profiles typical of lysosomes. No differences were found between the lipids of total control and affected livers, nor the fatty acid profiles of their phosphatidylcholine, phosphatidylethanolamine, or triglycerides. It is concluded that ovine ceroid lipofuscinosis is not a lipidosis, nor does the lipopigment arise from the abnormal peroxidation of lipids. Strong similarities between the lipopigment and the age pigment lipofuscin were noted.

Peroxisomes in several congenital syndromes (infantile refsum's disease, adrenoleukodystrophy, menkes' disease, batten's ceroid lipofuscinosis, GM1 gangliosidosis, a.c.)

Peroxisomes in several congenital syndromes (infantile refsum's disease, adrenoleukodystrophy, menkes' disease, batten's ceroid lipofuscinosis, GM1 gangliosidosis, a.c.)

Ceroid lipopigment: A morphometric study on its breakdown

Myocardium from English setters affected with ceroid lipofuscinosis (CCL) was maintained in culture for up to 14 days. Supplementing the culture medium with 0.2 mg/ml of N-acetyl-DL-homocysteine thiolactone (AHT) or with 5 μgml of dimethylaminoethyl-p-chlorophenoxyacetic acid (DPX) caused a significant reduction in number and an increase in size and volume density of lipopigment granules as determined by computer morphometric analysis. Electron microscopy showed changes in the consistency of the lipopigment in the form of vacuoles and pigment-body fragments. The antioxidant nordihydroguaiaretic acid (NDGA) caused an increase in number, size and volume density of the pigment granules. NN′-diphenyl-p-phenylenediamine (DPPD) and the enzyme horseradish peroxidase (HRP) may also produce similar changes when concentrations lower than 10 μg/ml and 80 μg/ml respectively are being used.

Dolichol and phosphorylated dolichol content of tissues in ceroid-lipofuscinosis.

Concentrations of free dolichol, total non-phosphorylated dolichol, and total phosphorylated dolichol were measured in autopsy specimens of brain and liver from ceroid-lipofuscinosis (CL) and control cases. Levels of non-phosphorylated dolichol, mainly as free dolichol, were increased approximately two-fold in late-infantile CL brain. No increase was observed for CL liver. In late-infantile and juvenile CL brain, the increase in phosphorylated dolichol was at least ten-fold, but no significant difference was found for CL liver. A substantial part of the increase in phosphorylated dolichol in CL brain appeared to be due to lipid-linked oligosaccharides. The results suggest that CL might involve a defect in the metabolism of dolichol-linked oligosaccharides.

Inherited disorders of pigmentation

Hermansky-Pudlak syndrome (HPS) is a rare autosomal recessive disorder causing oculocutaneous albinism, bleeding disorder and ceroid lipofuscinosis. Platelets from HPS patients are characterized by impaired secretion of dense (δ)-bodies (CD63). Meanwhile, there are ten known human HPS genes, each leading to a particular clinical HPS subtype (HPS1-HPS10).We report on two Turkish brothers showing typical HPS phenotype comprising oculocutaneous albinism and bleeding symptoms. Pathological bleeding time as well as platelet aggregometry analyses revealed impaired platelet function. The brothers demonstrated absence of platelet δ-granule secretion as measured by flow cytometry. Using molecular genetic analyses, a novel homozygous 1 bp-deletion in the HPS3 gene was identified in both brothers. In addition, the younger brother with HPS3 demonstrated psychomotoric retardation and cranial gliosis (magnetic resonance imaging, MRI). Array-CGH analysis revealed a de novo 0.482 Mb deletion on chromosome 17 which is not present in his brother and parents.In this study, we identified a novel 1 bp-deletion in the HPS3 gene causing HPS3 phenotype in two brothers. In patients with oculocutaneous albinism and increased bleeding symptoms platelet function should be analyzed. The identification of the molecular genetic defect allows the classification to a particular HPS subtype and is important for therapy and prognosis.

Dolichol and dolichyl phosphate levels in brain tissue from English setters with ceroid lipofuscinosis.

Human ceroid lipofuscinosis (CL) is an inherited disease marked by cerebromacular degeneration and early death. We have utilized the canine model to investigate the possible role of dolichol and dolichyl phosphate in the developmental pathology of CL. We found that while brain levels of dolichol increase with age in both affected and unaffected dogs, the amount in the diseased animal was similar to that in controls. Brain levels of dolichyl phosphate ranged from 20 to 35 micrograms/g in control dogs at all ages examined, but increased substantially during development in the affected dogs, a value of 113 +/- 24 micrograms/g (mean +/- SD) being obtained in the end-stage animals. In addition to the results obtained in the canine model, dolichyl phosphate levels in human brain tissues from a 5-year-old with late infantile CL and from a 19-year-old with juvenile CL were found to be 153 and 382 micrograms/g, respectively, compared with a control that assayed 26 micrograms/g. The preliminary findings with human tissues provide further evidence for an association of elevated brain dolichyl phosphate levels with CL. Whether the increase is primary or secondary remains to be determined.

Influence of N-acetylhomocysteine thiolactone on cultured retinal cells in canine neuronal lipofuscinosis. An ultrastructural study.

An SH- group donor drug, N-acetylhomocysteine thiolactone (0.2 mg/ml), was added to retinal cultures from an adult dog affected with ceroid lipofuscinosis (NCL). This substance was applied every 3rd day to the culture medium. After 1, 2 and 3 weeks in culture the cells were fixed for electron microscopy. Ultrastructurally the NCL-specific lipopigments revealed extensive fading out or vacuolation, revealing randomly arranged lamellar or 'fingerprint' membranes as well as structures resembling degenerated mitochondria. Additionally extensive fragmentation of the pigment bodies was present. The significance of these results are discussed.

Myoclonic, atonic, and absence seizures following institution of carbamazepine therapy in children

Five children, aged 3 to 11 years, treated with carbamazepine for epilepsy, had an acute aberrant reaction characterized by the onset of myoclonic, atypical absence and/or atonic (minor motor) seizures within a few days. When the carbamazepine was discontinued, two of the children returned to their former state very quickly, two had the minor motor seizures resolve in 3 and 6 months, and one had the seizures persist. The child in whom the seizures persisted was later found to have ceroid lipofuscinosis. The other children are doing well on other anticonvulsants.

Cutaneous ultrastructural diagnosis of ceroid-lipofuscinosis

A skin biopsy from a 6-year-old girl was examined by electron microscopy and the diagnosis of ceroid-lipofuscinosis (CL) was confirmed. This has not been reported in the dermatology literature, because of the absence of specific skin manifestations. However, as we have shown, various cells of skin, including fibroblasts, Schwann cells, eccrine serous cells, endothelial cells and smooth muscle cells, accumulated characteristic substances to produce the finger print pattern (FPP) or the curvilinear profile (CLP) which are thought to be diagnostic for CL. Cutaneous ultrastructural research in such cases is important as asymptomatic skin disorders may exist in other congenital storage diseases.

24 ULTRASTRUCTURAL FINDINGS OF SKIN AND PERIPHERAL NERVE BIOPSIES IN DEGENATIVE DISORDERS OF THE CENTRAL NERVE SYSTEM

In South America and, particularly in Brazil technical problems make difficult the performance of the majority of the lysosomal enzymes determinations and chemical analysis of central nervous system tissues. Thus, the correct diagnosis of most of the lyso-somial diseases is done by clinical and morphological data. Furthermore, the diagnosis of some degenerative diseases of the central nervous system whose biochemical background is unknown is based only on morphological data. This paper shows the ultrastructural findings in 70 consecutive skin and peripheral nerve biopsies from patients with a clinical history of a cerebral degenerative disorder. Positive results were obtained mainly in lysosomal diseases, i.e., the several forms of mucopolysaccharidosis, neurolipidosis and ceroid lipofuscinosis.

Ceroid Lipofuscinosis

Seven of ten patients with ceroid lipofuscinosis, including infantile, late infantile, juvenile, and atypical variants, presented with visual acuity loss. In the infantile case visual acuity loss was early and severe. Two patients with atypical variants did not develop ocular abnormalities. Children presenting with macular degeneration may have more extensive retinal involvement on electrophysiologic testing; neurologic evaluation may rule out the prodromal stages of lipofuscinosis.

Skin punch biopsies and lymphocytes in the diagnosis of lipidoses.

Skin punch biopsies and buffy coats of white blood cells were examined electron microscopically in patients suffering from a variety of storage diseases. No specific abnormalities could be detected in Gaucher's disease and adreno-leucodystrophy. While characteristic deposits were found in cutaneous nerves in globoid and metachromatic leucodystrophy, this method was deemed inferior to sural nerve biopsy. In gangliosidoses, on the other hand, pathognomonic membranous cytoplasmic bodies were common in axons of cutaneous nerves, and in generalized gangliosidoses marked vacuolation of many other cells was prominent. Specific deposits were found in various cells in skin punch biopsies and in lymphocytes of children suffering from ceroid lipofuscinoses, and in lymphocytes of their parents. This constitutes the easiest diagnostic laboratory procedure in such cases.

The Importance of Retinal Electrophysiology in Ceroid Lipofuscinosis

The two types of amaurotic idiocy, gangliosidosis and ceroid lipofuscinosis, are characterized by biochemical, ultrastructural, and electrophysiological differences. They are in fact two different clinical entities, both resulting from a congenital metabolic trouble.Six clinical cases of ceroid lipofuscinosis are presented and the diagnostic importance of the ERG is stressed.