Transcriptional Expression of Myelin Basic Protein in Oligodendrocytes Depends on Functional Syntaxin 4:a Potential Correlation with Autocrine Signaling

Myelination of axons by oligodendrocytes is essential for saltatory nerve conduction. To form myelin membranes, a coordinated synthesis and subsequent polarized transport of myelin components are necessary. Here, we show that as part of the mechanism to establish membrane polarity, oligodendrocytes exploit a polarized distribution of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) machinery components syntaxins 3 and 4, localizing to the cell body and the myelin membrane, respectively. Our data further reveal that the expression of myelin basic protein (MBP), a myelin-specific protein that is synthesized "on site" after transport of its mRNA, depends on the correct functioning of the SNARE machinery, which is not required for mRNA granule assembly and transport per se. Thus, downregulation and overexpression of syntaxin 4 but not syntaxin 3 in oligodendrocyte progenitor cells but not immature oligodendrocytes impeded MBP mRNA transcription, thereby preventing MBP protein synthesis. The expression and localization of another myelin-specific protein, proteolipid protein (PLP), was unaltered. Strikingly, conditioned medium obtained from developing oligodendrocytes was able to rescue the block of MBP mRNA transcription in syntaxin 4-downregulated cells. These findings indicate that the initiation of the biosynthesis of MBP mRNA relies on a syntaxin 4-dependent mechanism, which likely involves activation of an autocrine signaling pathway

Performance evaluation of family-based dispatching in small manufacturing cells

In many practical instances, the choice of whether to apply family-based dispatching or not can be decided per machine. The present paper explores the impact of the location of family-based dispatching, load variations between machines and routing of jobs on the flow time effect of family-based dispatching. These factors are explored in small manufacturing cells with and without labour constraints. An industrial case motivates the study. A simulation study is performed to assess the impact of these effects. The results show that shop-floor characteristics such as routing and load variation impact the decision where to locate family-based dispatching in manufacturing cells without labour constraints. By contrast, the effect of family-based dispatching is much less vulnerable to shop-floor characteristics in cells with labour constraints. Since workers are the bottleneck in these cells, it becomes less important at what machine the set-up time involving a worker is reduced. In general, there seems to be a trade-off between the positive effect of applying family-based dispatching at a (bottleneck) machine and the possible negative effect of the more irregular job arrivals at subsequent machines. The results further indicate that family-based dispatching is more advantageous in cells with labour constraints than in cells without labour constraints, when both types of manufacturing cells have comparable machine utilizations

Family based dispatching in manufacturing networks

Intrinsic to family based dispatching is the grouping of similar types of jobs in front of a machine for joint processing. Machine flow times may be improved in this way, as less time is spent on set-ups. Our observations in practice, however, suggest that family based dispatching may result in a bulky arrival pattern for successor manufacturing stages, thereby causing additional delay. So far, literature seems to neglect this effect. To explore this issue we develop queuing theoretical approximations of flow times for a simple two-stage shop. It appears that the optimal batch size for the shop is typically smaller than the optimal batch size for the batch machine. Furthermore, we propose extensions to existing dispatching rules by using information on successor stages. Existing and new extended rules are tested by an extensive simulation study. In line with the queuing theoretical analysis the outcomes indicate that exhaustive rules assuming batch size to be equal to family queue length - are clearly outperformed by non-exhaustive rules - allowing for smaller batches. Moreover, results show that the inclusion of local information on successor stages in rule decision making improves shop flow times

A New IT Architecture for the Supply Chain of Configurable Products

Part 5: Smart Production for Mass CustomizationInternational audienceMany ERP systems support configurable materials. Due to an ever increasing number of product variants the benefits of this approach are well understood. However, these implementations are not standardized. In this article we propose a new standard interface for the exchange of configuration data. This would lead to further benefits as systems as Advanced Planning systems could better use manufacturing flexibility while web shops as Amazon could easily integrate manufacturers of complex products with much reduced implementation effort

MAL Is a Regulator of the Recruitment of Myelin Protein PLP to Membrane Microdomains

In oligodendrocytes (OLGs), an indirect, transcytotic pathway is mediating transport of de novo synthesized PLP, a major myelin specific protein, from the apical-like plasma membrane to the specialized basolateral-like myelin membrane to prevent its premature compaction. MAL is a well-known regulator of polarized trafficking in epithelial cells, and given its presence in OLGs it was therefore of interest to investigate whether MAL played a similar role in PLP transport in OLGs, taking into account its timely expression in these cells. Our data revealed that premature expression of mCherry-MAL in oligodendrocyte progenitor cells interfered with terminal OLG differentiation, although myelin membrane formation per se was not impaired. In fact, also PLP transport to myelin membranes via the cell body plasma membrane was unaffected. However, the typical shift of PLP from TX-100-insoluble membrane domains to CHAPS-resistant, but TX-100-soluble membrane domains, seen in the absence of MAL expression, is substantially reduced upon expression of the MAL protein. Interestingly, not only in vitro, but also in developing brain a strongly diminished shift from TX-100 resistant to TX-100 soluble domains was observed. Consistently, the MAL-expression mediated annihilation of the typical membrane microdomain shift of PLP is also reflected by a loss of the characteristic surface expression profile of conformation-sensitive anti-PLP antibodies. Hence, these findings suggest that MAL is not involved in vesicular PLP trafficking to either the plasma membrane and/or the myelin membrane as such. Rather, we propose that MAL may regulate PLP's distribution into distinct membrane micro-domains that allow for lateral diffusion of PLP, directly from the plasma membrane to the myelin membrane once the myelin sheath has been assembled

Sulfatide-mediated control of extracellular matrix-dependent oligodendrocyte maturation

In the central nervous system, the extracellular matrix (ECM) compound laminin-2, present on developing axons, is essential in regulating oligodendrocyte (OLG) maturation. For example, laminin-2 is involved in mediating interactions between integrins and growth factors, initially localizing in separate membrane microdomains. The galactosphingolipid sulfatide is an important constituent of these microdomains and may serve as a receptor for laminin-2. Here, we investigated whether sulfatide interferes with ECM-integrin interactions and, in this manner, modulates OLG maturation. Our data reveal that disruption of laminin-2-sulfatide interactions impeded OLG differentiation and myelin-like membrane formation. On laminin-2, but not on (re)myelination-inhibiting fibronectin, sulfatide laterally associated with integrin alpha 6 in membrane microdomains. Sulfatide was partly excluded from membrane microdomains on fibronectin, thereby likely precluding laminin-2-mediated myelination. Anti-sulfatide antibodies disrupted integrin alpha 6-PDGF alpha R interactions on laminin-2 and induced demyelination in myelinated spheroid cultures, but intriguingly stimulated myelin-like membrane formation on fibronectin. Taken together, these findings highlight the importance of laminin-sulfatide interactions in the formation of functional membrane microdomains essential for myelination. Thus, laminin-sulfatide interactions might control the asynchronous localized differentiation of OLGs, thereby allowing myelination to be triggered by axonal demand. Given the accumulation of fibronectin in multiple sclerosis lesions, the findings also provide a molecular rationale for the potential of anti-sulfatide antibodies to trigger quiescent endogenous OLG progenitor cells in axon remyelination. GLIA 2014;62:927-94

Interference of poly(ethylene glycol)-lipid analogues with cationic-lipid-mediated delivery of oligonucleotides; role of lipid exchangeability and non-lamellar transitions.

Cationic liposomes are applied to transfer oligonucleotides (ODNs) into cells to regulate gene expression for gene therapeutic or cell biological purposes. In vivo, poly(ethylene glycol) (PEG)-lipid derivatives are employed to stabilize and prolong the circulation lifetime of nucleic acid-containing particles, and to improve targeting strategies. In this study, we have studied the effects of PEG-lipid analogues, i.e. PEG coupled to either phosphatidylethanolamine (PE) or ceramide, on cationic-lipid-DNA complex ('lipoplex') assembly and the mechanism of cationic-lipid-mediated delivery of ODNs in vitro. Inclusion of 10 mol% PEG-PE in ODN lipoplexes inhibited their internalization in Chinese hamster ovary cells by more than 70%. The intracellular fraction remained entrapped in the endosomal/lysosomal pathway, and no release of ODNs was apparent. Similar observations were made for complexes prepared from liposomes that contained PEG-ceramides. Interestingly, delivery resumed when lipoplexes had been externally coated with PEG-ceramides. In this case, the kinetics of delivery were dependent on the length of the ceramide acyl chain, consistent with a requirement for the PEG-lipid to dissociate from the complex. Moreover, although the chemical nature of the PEG-ceramides distinctly affected the net internalization of the complexes, impediment of delivery was largely related to an inhibitory effect of the PEG-lipid on the release of ODNs from the endosomal compartment. Cryo-electron microscopy and small-angle X-ray scattering revealed that the PEG-lipids stabilize the lamellar phase of the lipoplexes, while their acyl-chain-length-dependent transfer from the complex enables adaptation of the hexagonal phase. Within the endosomal compartment, this transition appears to be instrumental in causing the dissociation and cytosolic release of the ODNs for their nuclear homing