Myelin Basic Protein Expression In Oligodendrocytes Research Articles

Hericium erinaceus mycelium and its small bioactive compounds promote oligodendrocyte maturation with an increase in myelin basic protein

Oligodendrocytes (OLs), myelin-producing glia in the central nervous system (CNS), produce a myelin extension that enwraps axons to facilitate action potential propagation. An effective approach to induce oligodendrogenesis and myelination is important to foster CNS development and promote myelin repair in neurological diseases. Hericium (H.) erinaceus, an edible and culinary-medicinal mushroom, has been characterized as having neuroprotective activities. However, its effect on OL differentiation has not yet been uncovered. In this study using oligodendrocyte precursor cell (OPC) cultures and an ex vivo cerebellar slice system, we found that the extract from H. erinaceus mycelium (HEM) not only promoted the differentiation of OPCs to OLs in the differentiation medium, but also increased the level of myelin basic protein (MBP) on neuronal fibers. Moreover, daily oral administration of HEM into neonatal rat pups for 7 days enhanced MBP expression and OLs in the corpus callosum of the postnatal rat brain. The effect of HEM-derived bioactive compounds, the diterpenoid xylosides erinacine A (HeA) and HeC and a sesterterpene with 5 isoprene units called HeS, were further evaluated. The results showed that HeA and HeS more potently stimulated MBP expression in OLs and increased the number of OLs. Moreover, overlap between MBP immunoreactivity and neuronal fibers in cultured cerebellar tissue slices was significantly increased in the presence of HeA and HeS. In summary, our findings indicate that HEM extract and its ingredients HeA and HeS display promising functional effects and promote OL maturation, providing insights into their potential for myelination in neurodevelopmental disorders.

Effects of propofol on myelin basic protein expression in oligodendrocytes of SD rats at different developmental stages

To investigate the effect of propofol on myelin basic protein (MBP) expression in oligodendrocytes of SD rats at different developmental stages. This study was conducted in 3?, 7?, 14? and 21?day?old SD rats (40 in each age group). In each group, the rats were randomized equally into control group and experimental group, and in the control group, the rats received an intraperitoneal injection of 25 mg/kg medium?long?chain fat emulsion followed by injections at a half dose every 20 min for 8 h; the rats in the experimental group were given injections of propofolmedium (at the initial dose of 25 mg/kg) in the same manner. The transcriptional levels of MBP and caspase?3 in the brain tissues were detected by qRT?PCR, and the protein expression of MBP was with Western blotting and immunehistochemistry. Compared with those in the control groups, the expression of MBP mRNA was significantly down?regulated while caspase?3 mRNA was up?regulated in 3?, 7? and 14?day?old rats in the experimental groups (P<0.05). The protein expression of MBP in 7? and 14?day?old rats was significantly decreased in the experimental groups compared with the control groups (P<0.05). The expression of caspase?3 mRNA or MBP protein in 21?day?old rats showed no significant difference between the two groups (P>0.05). Propofol can down?regulate the expression of MBP at both the mRNA and protein levels in SD rats, especially in those at 7 and 14 days of age.

Heterogeneous Nuclear Ribonucleoprotein (hnRNP) F Is a Novel Component of Oligodendroglial RNA Transport Granules Contributing to Regulation of Myelin Basic Protein (MBP) Synthesis

Myelin basic protein (MBP) is a major component of central nervous system (CNS) myelin. The absence of MBP results in the loss of almost all compact myelin in the CNS. MBP mRNA is sorted into RNA granules that are transported to the periphery of oligodendrocytes in a translationally inactive state. A central mediator of this transport process is the trans-acting factor heterogeneous nuclear ribonucleoprotein (hnRNP) A2 that binds to the cis-acting A2-response element in the 3'UTR of MBP mRNA. Recently, we found that activation of the Src family nonreceptor tyrosine kinase Fyn in oligodendrocytes leads to phosphorylation of hnRNP A2 and to increased translation of MBP mRNA. Here, we identify the RNA-binding protein hnRNP F as a novel component of MBP mRNA transport granules. It is associated with hnRNP A2 and MBP mRNA in cytoplasmic granular structures and is involved in post-transcriptional regulation of MBP expression. Fyn kinase activity results in phosphorylation of hnRNP F in the cytoplasm and its release from MBP mRNA and RNA granules. Our results define hnRNP F as a regulatory element of MBP expression in oligodendrocytes and imply an important function of hnRNP F in the control of myelin synthesis.

Rescuing qkvDysmyelination by a Single Isoform of the Selective RNA-Binding Protein QKI

Alternative splicing of the qkI transcript generates multiple isoforms of the selective RNA-binding protein QKI, which play key roles in controlling the homeostasis of their mRNA targets. QKI deficiency in oligodendrocytes of homozygous quakingviable (qkV/qkV) mutant mice results in severe hypomyelination, indicating the essential function of QKI in myelinogenesis. However, the molecular mechanisms by which QKI controls myelination remain elusive. We report here that QKI-6 is the most abundant isoform in brain and is preferentially reduced in the qkV/qkV mutant during normal myelinogenesis. To test whether QKI-6 is the predominant isoform responsible for advancing CNS myelination, we developed transgenic mice that express Flag-QKI-6 specifically in the oligodendroglia lineage, driven by the proteolipid protein (PLP) promoter. When introduced into the qkV/qkV mutant, the QKI-6 transgene rescues the severe tremor and hypomyelination phenotype. Electron microscopic studies further revealed that the Flag-QKI-6 transgene is sufficient for restoring compact myelin formation with normal lamellar periodicity and thickness. Interestingly, Flag-QKI-6 preferentially associates with the mRNA encoding the myelin basic protein (MBP) and rescues MBP expression from the beginning of myelinogenesis. In contrast, Flag-QKI-6 binds the PLP mRNA with lower efficiency and has a minimal impact on PLP expression until much later, when the expression level of QKI-6 in the transgenic animal significantly exceeds what is needed for normal myelination. Together, our results demonstrate that QKI-6 is the major isoform responsible for CNS myelination, which preferentially promotes MBP expression in oligodendrocytes.

Sox10 acts as a tissue‐specific transcription factor enhancing activation of the myelin basic protein gene promoter by p27Kip1 and Sp1

Myelin basic protein (MBP) is one of the major components of the myelin sheath that insulates axons. In the central nervous system, MBP is synthesized by differentiating oligodendrocytes. The expression of MBP in oligodendrocytes is regulated mainly at the transcriptional level. The Sp1 family of transcription factors has been shown to be important in the regulation of many genes. Binding of Sp1 to the GC box in the proximal MBP promoter has been shown to be indispensable for the activation of MBP gene expression. Previous results from our laboratory have shown that the increase in p27Kip1 that accompanies oligodendrocyte differentiation is paralleled by an increase in Sp1. We also have shown that the increase in MBP expression resulting from elevated p27Kip1 levels is mediated through Sp1 and that this effect occurs specifically in oligodendrocytes. In this study, we found that increased expression of p27Kip1 together with the nervous-system-specific transcription factor Sox10 can activate the MBP promoter even in nonoligodendrocyte cells. This indicates that Sox10 confers cell type specificity on the expression of MBP. Both Sp1 and Sox10 can enhance MBP promoter activity when expressed alone. Cotransfection of plasmids encoding Sp1 and Sox10 induces increased activation of the MBP promoter over expression of either transcription factor alone. This effect is not limited to oligodendrocyte cell lines, in that Sp1 and Sox10 can also synergistically activate the MBP promoter when expressed in NIH3T3 fibroblasts. Mutation of the Sp1 binding sites in the MBP promoter eliminates Sox10 stimulated activation, suggesting that the MBP promoter is activated, at least in part, through protein-protein interactions between Sp1 and Sox10.

MyEF-3, a Developmentally Controlled Brain-Derived Nuclear Protein Which Specifically Interacts with Myelin Basic Protein Proximal Regulatory Sequences

Myelin basic protein (MBP) gene contains the upstream regulatory sequence that confers cell type- and stage-specific transcription to MBP expression in oligodendrocytes during brain development. The MB1regulatory motif located between −14 to −50 with respect to transcription start site binds to a brain derived nuclear protein and plays an important role in transcriptional activation of the MBP promoter in transfection assay. Here, we report the isolation of a recombinant cDNA clone, termedmyelinexpressionfactor-3,(MyEF-3) from a mouse brain expression library that encodes a novel protein which interacts with the MBP MB1domain. Computer assisted evaluations of the MyEF-3 sequence revealed several interesting features including four sites for phosphorylation by casein kinase II, a transmembrane domain at the N-terminus, a nuclear localization signal and a Zinc finger domain at the carboxyl terminal. Results from Western and band shift assays indicate that MyEF-3 binds efficiently to double-stranded MB1as well as the single-stranded non-coding strand of MB1. The use of short DNA fragments encompassing the nucleotide base substitutions across the MB1 domain in competition band shift assay revealed that the ten nucleotide sequence, 5′-GCCTGTCTTT-3′ is important for binding of MyEF-3 to DNA. Results from Northern blot studies demonstrate that expression of MyEF-3 is restricted to brain and developmentally regulated during brain maturation. The biological importance of MyEF-3 in the cell type- and stage-specific expression of MBP during brain development is discussed.

Actions of steroid hormones and growth factors on glial cells of the central and peripheral nervous system

Primary cultures of oligodendrocytes and astrocytes and purified cultures of Schwann cells were prepared respectively from forebrain and sciatic nerves of newborn rats. The effects of steroid hormones and growth factors on glial cell growth and on the production of myelin-specific proteins and lipids were investigated. Progesterone (P, 100 nM) decreased the proliferation of glial cells of the central nervous system. This inhibitory effect of P was abolished by the simulatenous adminnistration of the antagonist RU486, thus suggesting a receptor-mediated action of the hormone. The expression of myelin-specific proteins, including the myelin basic protein (MBP) and the 2′,3′-cyclic nucleotide-3′-phosphodiesterase (CNPase), and of a myelin-specific lipid, glactocerebroside (Gal C), was also measured during cell differentiation under different hormonal conditions. The expression of MBP in oligodendrocytes was increased by P, and this effect was not blocked by RU486. The combined application of P and insulin promoted a synergistic stimulation of MBP expression. Insulin, by itself, also increased the number of MBP-positive oligodendrocytes in culture. The effects of P and insulin appeared to be selective as dexamethasone, dehydroepiandrosterone, pregnanolone and epidermal growth factor (EGF) had no effect. Only estradiol (E 2, 500 nM) increased the number of MBP-immunoreactive cells, but in contrast to P, only a small synergism between E 2 and insulin on MBP expression was observed. The expression of CNPase, another myelin-specific protein, was also increased by P and, here again, a synergy between P and insulin could be observed. In contrast, the expression of Gal C, a myelin-specific lipid, was not modified by P or other steroid hormones. Moreover, the increase in Gal C-positive cells observed in response to insulin alone was not further potentiated by P. Glial cells of the peripheral nervous system, namely Schwann cells, are also sensitive to steroid hormones. Schwann cells contain estrogen receptors, and E 2 stimulates their proliferation in the presence of forskolin or dibutyryl cyclic AMP (dbcAMP). The mitogenic effect of E 2 was abolished by the pure antiestrogen ICI-164,384. Insulin, at micromolar concentration, also stimulated Schwann cell growth when forskolin or dbcAMP were present in the culture medium. The mitogenic effect of insulin was mediated by insulin-like growth factor I (IGF-I) receptors. Indeed, at a physiological nanomolar concentration, IGF-I but not insulin or IGF-II, increased the proliferation of Schwann cells in synergy with forskolin. In addition, Schwann cells express receptors for IGF-I.