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N Herschkowitz

Publications and source records attributed to N Herschkowitz.

At least 73 records · Page 4Linked to original sources

Viral tropisms in mouse brain cell cultures.

Fourteen-day-old cultures of dissociated newborn mouse brain cells were infected separately with different strains of vaccinia virus and a strain of measles virus. Using the indirect immunofluorescence technique we found that under the experimental conditions in these cultures both measles and the neurotropic strain of vaccinia infected oligodendrocytes whereas the dermatropic strain of vaccinia did not. Astrocytes were neither infected by vaccinia strains nor by measles virus.

Animals↗

Oligodendrocytes of jimpy mice express galactosylceramide: an immunofluorescence study on brain sections and dissociated brain cell cultures.

Brain sections and dissociated brain cell cultures of jimpy mouse (jp) were investigated for the presence of galactosylceramide (GC) by indirect immunofluorescence. Optic nerve and corpus callosum sections of 26-day-old jp exhibited many GC-positive cells. The GC staining pattern was similar in jp and normal cultures of the same age. These data suggest that the previously observed decreased amount of GC in jp brain is due to the inability of jp oligodendroglia to properly deposit GC in the myelin, while its synthesis is possible.

Animals↗

Relative changes in the amount of galactocerebroside during development as observed in mouse brain cell cultures.

Relative changes in the amount of galactocerebroside (GC) during development were measured in mouse brain cell cultures at different stages of development. For this purpose we used an 125I-labelled protein A indirect assay modified in the respect that the total amount of cellular proteins was evaluated before counting the radioactivity. The amount of GC greatly increased between the 10th and the 14th day of culture, then a steady state was reached between the 14th and the 20th day of culture. This change correlates well with the dynamics of the number of oligodendrocytes we observed earlier. These data suggests that the increase of the GC amount in culture during development corresponds to the increase in the number of GC-positive oligodendrocytes rather than to the increase in the number of GC molecules per cell.

Animals↗

Expression of antigenic markers during the development of oligodendrocytes in mouse brain cell cultures.

The expression of myelin basic protein (MBP) and galactocerebroside (GC), two antigenic markers for oligodendrocytes, was checked on 7-, 14-, 21- and 28-day-old dissociated mouse brain cell cultures (BCC) by using the indirect immunofluorescence method with double staining. The number of GC positive cells increased between the 7th and the 14th day of culture before a steady state was reached. In contrast to this, the MBP-positive cells appeared only on the 14th day of culture, and their number increased with the age of the culture. In double staining, the serum produced against isolated oligodendrocytes shows the same picture as the anti-GC serum, while only a part of GC-positive cells showed also the presence of MBP. Our data suggest that the GC appears very early on the membrane of the oligodendrocytes during development while cells exhibiting both GC and MBP probably represent a more differentiated oligodendrocyte population.

Animals↗

Immunological properties of bulk isolated mouse oligodendrocytes.

Mouse oligodendrocytes were isolated in bulk. These cells were 95% galactocerebroside-positive. Rabbit anti-mouse oligodendrocyte antisera reacted in radioimmunoassay and in indirect immunofluorescence with mouse oligodendrocytes in suspension or in culture. The activity of the sera was not blocked by adsorption with galactocerebroside or by preincubation of cells with anti-galactocerebroside serum, although it cross-reacted with galactocerebroside in a serological test. The staining was not impaired by the pretreatment of cells with trypsin, neuraminidase or acetone, nor by adsorption of serum with myelin basis protein.

Animals↗

Synthesis of lipids in mouse brain cell cultures during development.

Several metabolic activities in dissociated cultures of newborn mouse brain were compared to the situation in vivo. The developmental activity pattern of cerebroside-sulfotransferase, cyclic nucleotide phosphohydrolase, and beta-hydroxy-beta-methyl glutaryl-coenzyme A-reductase and the synthesis and deposition of sulfatide and cholesterol in culture were estimated. The enzyme activity patterns in vivo and in culture are the same. Since the cultures show very little myelin formation, the parallel increase of enzyme activities necessary for myelination in vivo and in culture suggest the existence of intrinsic factors regulating the biochemical differentiation. In addition, the formation of the products, determined in culture, follows the patterns of the enzyme activities. Dissociated brain cell cultures are therefore a valid model for the study of biochemical parameters related to the synthesis of brain lipids during development.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

Distribution of cytoskeletal elements in cultured skin fibroblasts of patients with Duchenne's muscular dystrophy.

Cultured fibroblasts from patients suffering from Duchenne's Muscular Dystrophy were examined by indirect immunofluorescent techniques using antibodies against actin, myosin, tubulin, and intermediate-sized filaments. The cells display normal patterns of microfilamentous bundles (stress fibres), microtubules, and intermediate-sized filaments suggesting a normal organization of these cytoskeletal structures.

Actins↗

Immunohistochemical localization of renin in mouse brain.

Immunohistochemical studies of mouse brain by the peroxidase-antiperoxidase method using monospecific antimouse renin antibodies has revealed the intracellular localization of renin in stellate and small ovoid cells. Renin-containing ovoid cells were observed in the spinal cord, medulla oblongata, pons, granular layers of the cerebellum, deep cerebellar region, and the lamina terminalis; whereas immunostainable stellate cells were found in the cerebral cortex, hippocampus, dentate gyrus and septum. Intracellular localization of renin rather than intravascular localization supports an endogenous origin of this enzyme in the brain. Wide distribution in different types of cells suggests different types of regulatory mechanisms. Double immunostaining with antigalactocerebroside and antirenin antibodies indicates the presence of renin in oligodendrocytes.

Animals↗

Adrenomyeloneuropathy. A protracted, pseudosystematic variant of adrenoleukodystrophy.

Histopathological, immunocytochemical, and electron microscopical investigations were carried out in a man with a protracted history of spastic paraparesis, adrenal insufficiency and hypogonadism. Pathological findings were identical with those of the few previously reported cases of adrenomyeloneuropathy (AMN) including cytoplasmic lamellar inclusions consisting of two parallel 2.5 nm leaflets separated by a clear space of variable extent, in the brain, spinal roots, adrenal gland, and interstitial cells of the testis. No inclusions could be found in oligodendrocytes. In brain macrophages they are thought to represent breakdown products of pathological myelin stored in lysosomes, whereas in other localizations they might be an expression of the primary metabolic defect of the cell. Special attention was paid to the pseudosystematic type of fiber tract degeneration in the spinal cord. The dying-back pattern of axonal destruction was interpreted as a possible result of the multisegmental demyelination observed in these tracts. All known hormones could be localized in the pituitary by immunocytochemistry. Corticotrophs and gonadotrophs were numerous. The structural damage of the adrenal cortex and the interstitial cells of the testis is, therefore, considered to result from the inborn error of metabolism on the one hand and from an enhanced stimulation exerted by ACTH and gonadotrophins on the other.

Adrenal Glands↗

Progressive somatofugal activation of cerebroside sulfotransferase in the developing chick optic fibers.

A myelination parameter of the chick optic pathway has been investigated: the activity of cerebroside sulfotransferase. Evidence is presented for a somatofugal progression of the enzyme activity peak along the optic nerve, chiasm, optic tract, tectum anterior, and tectum posterior. The enzyme activity appears on the fifteenth embryonic day and reaches a maximum in the nerve two days before hatching. One day after hatching, the peak is found in the chiasm, and three days after that in the optic tract. The peak is found in the tectum anterior on the tenth day. The results suggest a somatofugal progression of myelination.

Animals↗

Abnormal metabolism of 35SO4-sulfatide in jimpy brains expressed in brain organotypic cultures.

Cerebroside-sulfotransferase (CST), creatine-phosphokinase (CPK), and 3-hydroxy-3-methylglutaroyl CoA (HMG CoA) reductase activity, protein, and DNA content were measured in an easy-to-perform organotypic culture system of newborn normal and jimpy brains. The defective sulfatide synthesis which has been shown in vivo in jimpy brains could also be demonstrated in organ cultures of jimpy mice in the form of lowered CST activity in the homogenate as well as reduced 35SO4 incorporation into 35SO4-sulfatide. HMG CoA reductase was reduced to 60% of that found in 16-day-old normal cultures, similar to the findings in vivo. DNA of jimpy cultures was significantly lower than that in normal cultures, suggesting the possibility of an arrest in the differentiation or increased cellular death of presumptive oligodendrocytes, as was found in vivo. Organ cultures of jimpy mouse brain can serve as an appropriate model for further study of the primary defect in this animal mutant.

Animals↗

Prenatal mucopolysaccharidosis II (Hunter): a pathogenetic study.

A prenatal diagnosis of Mucopolysaccharidosis II (M. Hunter) was made early in a pregnancy at risk in a family with one affected child. An affected fetus was diagnosed on the basis of an abnormal incorporation and degradation of 35SO4 in 35SO4-labeled mucopolysaccharides in cultured amniotic cells. Dermatan sulfate and heparin sulfate concentrations in the supernatant of the amniotic fluid were high. In the aborted fetus, the diagnosis could be confirmed by 35SO4 incorporation studies in the cultured fibroblasts and in cultured brain cells as well as by the deficiency of the specific enzyme activity (iduronide sulfate sulfatase) in the organs of the fetus. beta-Galactosidase was in the low normal range in liver and spleen but significantly reduced in brain. Under electron microscopy, the mesenchymal cells of liver and spleen showed lysosomal storage of material, presumably mucopolysaccharides, in excess of normal. In the neurons of the spinal ganglia and spinal cord, "Zebra bodies" in statu nascendi were observed.

Female↗

Theophylline reduces the activity of cerebroside-sulfotransferase, a key enzyme in myelination, in cell cultures from newborn mouse brain.

Theophylline, a drug used in neonatology for the treatment of apnea, affects cholesterol synthesis if administered in concentrations of 10(-4) M (a concentration found in serum of treated patients) for 24 hr to dissociated brain cell cultures. The rate-limiting enzyme of cholesterol synthesis, beta-hydroxy-beta-methylglutaryl-coenzyme A reductase (EC 1.1.1.34), is lowered to 45% 48 hr after removal of theophylline. At the same time, cholesterol content of the cells is lowered to 73%. Inasmuch as the phospholipid content of the cells remains stable, the treatment changes the cholesterol phospholipid ratio. Concomitant to this effect, the activity of cerebroside-sulfotransferase (EC 2.8.2.11) is lowered to 60% of control values. We postulate that these two effects are linked to each other by means of modulation of the cerebroside-sulfotransferase activity by membrane lipids.

Animals↗

Influence of reduced cholesterol synthesis on the activity of cerebroside sulfotransferase in cultured glioblastoma cells treated with estradiol.

Cultured glioblastoma cells were inoculated with estradiol in concentrations of 0.5--10 microliter/ml medium in order to check the effect of this hormone on the activity of cerebroside sulfotransferase, an enzyme whose activity is strongly related to myelination. Thereby we could show that the cerebroside-sulfotransferase activity increases to a value of 200% of normal. Concomitant to this effect, the cholesterol content of the membrane bearing cerebroside sulfotransferase activity decreases to 60% of normal. The effect is fully reversible: after 48 h, cholesterol synthesis as well as cerebroside sulfotransferase activity reach normal values again. We suggest that cerebroside sulfotransferase activity is modulated by the changing cholesterol/phospholipid ratio in the cells during the inoculation period.

Animals↗