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S M Staugaitis

Publications and source records attributed to S M Staugaitis.

30 records · Page 2Linked to original sources

The ectopic expression of myelin basic protein isoforms in Shiverer oligodendrocytes: implications for myelinogenesis.

The myelin basic proteins (MBPs) are a set of peripheral membrane polypeptides that are required for the compaction of the major dense line of central nervous system myelin. We have used primary cultures of oligodendrocytes from MBP-deficient shiverer mice as host cells for the expression by cDNA transfection of each of the four major MBP isoforms. The distributions of the encoded polypeptides were studied by immunofluorescence and confocal microscopy and compared with patterns of MBP expression in normal mouse oligodendrocytes in situ and in culture. The exon II-containing 21.5- or 17-kD MBPs were distributed diffusely in the cytoplasm and in the nucleus of the transfectants, closely resembling the patterns obtained in myelinating oligodendrocytes in 9-d-old normal mouse brains. By contrast, the distribution of the 14- and 18.5-kD MBPs in the transfectants was confined to the plasma membrane and mimicked the distribution of MBP in cultures of normal adult oligodendrocytes. Our results strongly suggest that the exon II-containing MBPs are expressed first and exclusively during oligodendrocyte maturation, where they may play a role in the early phase of implementation of the myelination program. In contrast, the 14- and 18.5-kD MBPs that possess strong affinity for the plasma membrane are likely to be the principle inducers of myelin compaction at the major dense line.

Animals↗

Expression of the oligodendrocyte marker 2'3'-cyclic nucleotide 3'-phosphodiesterase in non-glial cells.

The 46 kD isoform of the 2'3'-cyclic nucleotide 3'-phosphodiesterase (CNPI) was expressed in HeLa cells by transfection of its cDNA clone. The distribution of this polypeptide as mapped by indirect immunofluorescence and conventional epifluorescence microscopy appeared diffuse and generally uniform throughout the cytoplasm. Confocal microscopic imaging and analysis of pseudocolored images confirmed this distribution but also revealed that there was a high concentration of CNPI near the plasma membrane of the cell. This pattern is very similar to that observed by immunoelectronmicroscopy of myelinating oligodendrocytes (Trapp et al.: J Neurochem 51:859-868, 1988; Braun et al.: J Neurosci 8: 3057-3066, 1988). These results suggest that CNP may interact with a membrane-associated molecule that is not unique to oligodendrocytes.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

Protein zero of peripheral nerve myelin: biosynthesis, membrane insertion, and evidence for homotypic interaction.

Protein zero (P0), an integral membrane glycoprotein synthesized by Schwann cells, is the major glycoprotein of peripheral nerve myelin. The predicted disposition of P0 with respect to the membrane bilayer postulates the existence of extracellular and intracellular domains, that mediate compaction of the myelin lamellae. We used in vitro translations programmed with sciatic nerve mRNA and cells transfected with a P0 cDNA construct to study the biosynthesis and topology of P0 in the bilayer. The behavior of P0 at the cell surface, when expressed under physiological conditions, was also examined. We have verified the topological predictions of an earlier model, derived from analysis of a P0 cDNA, and provide evidence that the extracellular domain of P0 mediates homotypically cell-cell interactions in the transfectants.

Amino Acid Sequence↗

Expression of myelin basic protein isoforms in nonglial cells.

The myelin basic proteins (MBPs) mediate the cytoplasmic apposition of the oligodendrocyte plasma membrane to form the major dense line of central nervous system myelin. Four major isoforms of murine MBP, obtained by alternative splicing of seven exons from a single primary transcript, display distinct developmental profiles. We expressed these major MBPs individually in HeLa cells and mapped their distributions by immunofluorescence and confocal microscopy. The 14- and 18.5-kD MBPs that are the predominant forms in compact myelin distributed primarily in the perinuclear regions of the cell in configurations highly suggestive of close association with membranes. We infer that these MBP isoforms possess strong, nonspecific membrane-binding properties that have been adapted by the oligodendrocyte to mediate compaction of the sheaths of plasma membrane that form myelin. In contrast, the 17- and 21.5-kD isoforms distributed diffusely in both the cytoplasm and the nucleoplasm and often accumulated within the nucleus. This distribution can be correlated with the presence of the peptide segment encoded by exon II, which is unique to these isoforms. The physiological significance of the nuclear targeting displayed by the 17- and 21.5-kD MBP isoforms in HeLa cells remains to be determined.

Amino Acid Sequence↗

Physiologic properties of myelin proteins revealed by their expression in nonglial cells.

The transfection paradigm described herein can be used to investigate the functional properties of individual nervous system proteins in ways that have not been explored before. In particular, observations on the "structural" proteins of myelin are being made that have already yielded certain unique insights into the physiologic properties of these polypeptides. The ease with which site-directed mutagenesis procedures can be applied to these systems should eventually enable us to define with great precision the "functional domains" within each myelin protein.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

Intra-blood-brain barrier IgG synthesis in cerebral cysticercosis.

Patients with central nervous system cysticercosis show elevated binding of cerebrospinal fluid (CSF) IgG to homogenized cysticercus. To determine whether any of the CSF IgG was the result of de novo intra-blood-brain barrier (BBB) synthesis, CSF and serum samples from six patients were examined for elevated rate of synthesis and oligoclonal bands. Five of the six patients had increased intra-BBB IgG synthesis rate and four of these patients also had oligoclonal IgG bands present in the CSF that were absent in the serum. These results demonstrate intra-BBB IgG synthesis similar to that observed in other infectious and inflammatory diseases of the central nervous system.

Blood-Brain Barrier↗

The basis of intra-blood-brain-barrier IgG synthesis.

We hold that the intra-blood-brain-barrier (BBB) IgG synthesis (SYN) rate can be quantitated reliably and validly. Although several formulas distinguish patients with multiple sclerosis from normal controls equally well, only the SYN rate formula has been validated in humans using isotopic tracer techniques. Our formula for the IgG SYN rate is reducible to Reiber's formula; therefore, both can be used to quantify the IgG SYN rate in a manner not possible using the IgG index. Although our SYN rate formula has been validated for a modest range of BBB abnormalities (cerebrospinal fluid/serum albumin ratios), there is evidence to suggest that it may be used even in patients having severe BBB damage. We question the acceptance of unique cerebrospinal fluid IgG bands as indisputable evidence of intra-BBB IgG SYN in the presence of a modest to severely damaged BBB. Finally, the utility of quantitation and detection of intra-BBB IgG SYN by standardized methods in a group of asymptomatic, normal individuals compared with a group of patients with clinical definite multiple sclerosis is presented.

Blood-Brain Barrier↗

Azathioprine and steroids are not more effective in decreasing multiple sclerosis intra-blood-brain-barrier IgG synthesis than steroids alone.

Intra-blood-brain-barrier IgG synthesis rates and oligoclonal IgG banding patterns were examined in 9 patients with multiple sclerosis who were treated with azathioprine and steroids for 2 to 4.5 years. The IgG synthesis rates of 5 patients were significantly decreased from the pretreatment mean values 1 month after treatment, and their synthesis rates remained at the decreased levels throughout treatment. However, among the remaining 4 patients, the rates exceeded the pretreatment means. This continuous suppressive effect of the combined azathioprine and steroids upon the IgG synthesis rate was similar to that of steroids, suggesting that azathioprine and steroids in combination were not more effective in reducing intra-blood-brain-barrier IgG synthesis than steroids alone. Oligoclonal IgG patterns in all cerebrospinal fluid samples were not significantly altered during the study.

Adrenocorticotropic Hormone↗

The current status of multiple sclerosis intra-blood-brain-barrier IgG synthesis.

The status of the central nervous system as an immunologic organ synthesizing IgG in several neurological diseases has been established convincingly in the past two decades. The measurement of intra-BBB IgG synthesis has been accomplished. It has been demonstrated that this synthesis is a potential marker for an abnormal immunologic process within the CNS which if manipulated may be a quantifiable aspect of disease modulation. Modulation of intra-BBB IgG synthesis is feasible, but there is no evidence yet that modulation of this synthesis is an index of effective disease control in MS. Only ACTH and corticosteroids lead to a significant and lasting depression of intra-BBB IgG synthesis, even though clonal eradication is not achieved. Based on our immunopharmacologic studies in MS we have proposed an immunokinetic model. The qualitative analysis of the specificity of IgG synthesized intra-BBB in MS has not led to the etiology of MS, but utilization of quantitative methods may. Questions raised by this review with suggested experiments to advance further our understanding of intra-BBB IgG synthesis as it relates to the etiopathogenesis of MS are included.

Antibody Formation↗

Hypothesis: multiple sclerosis cerebrospinal fluid IgG multiple bands are derived from one or a few IgG secretor cell clones.

Analytical IEF, immunofixation, and silver nitrate staining of mouse monoclonal, human myeloma, and multiple sclerosis (MS) oligoclonal IgG result in multiple bands. Because the IgG secreted by mouse hybridoma and multiple myeloma are the products of single clones of cells, it is reasonable to propose that MS CSF IgG multiple bands may be derived from one or a small number of clones of IgG secretors.

Antibody Specificity↗

Correlation of multiple sclerosis (MS) IgG fractionated by capillary isotachophoresis (ITP) and by isoelectric focusing polyacrylamide gel electrophoresis (IEF-PAGE).

Both IEF-PAGE and ITP have been used in the analysis of MS CSF IgG. To directly compare the two techniques we separated MS CSF IgG by IEF-PAGE, excised 1cm fractions of the gel, extracted and concentrated the IgG, and then analyzed each fraction by ITP. Each IEF fraction, with increasing isoelectric point, showed increasingly longer retention times on ITP, but there was a more detailed separation of proteins by ITP and the fractions overlapped. We conclude that ITP and IEF separate IgG into corresponding patterns.

Blood-Brain Barrier↗