Promoting myelin repair is one of the most promising therapeutic avenues in the field of myelin disorders. In future clinical trials, evaluation of remyelination will require a reliable and quantifiable myelin marker to be used as a surrogate marker. To date, MRI assessment lacks specificity for evaluating the level of remyelination within the brain. Here, we describe 1,4-bis(p-aminostyryl)-2-methoxy benzene (BMB), a synthesized fluorescent molecule, that binds selectively to myelin both ex vivo and in vivo. The binding of BMB to myelin allows the detection of demyelinating lesions in an experimental autoimmune encephalitis model of demyelination and allows a mean for quantifying myelin loss in dysmyelinating mutants. In multiple sclerosis brain, different levels of BMB binding differentiated remyelination in shadow plaques from either demyelinated lesions or normal-appearing white matter. After systemic injection, BMB crosses the blood-brain barrier and binds to myelin in a dose-dependent and reversible manner. Finally, we provide evidence that 11 C-radiolabeled BMB can be used in vivo to image CNS myelin by positron-emission tomography in baboon. Our results provide a perspective for developing a brain myelin imaging technique by positron-emission tomography.multiple sclerosis ͉ remyelination ͉ leukodystrophy M yelin is a unique structure in the nervous system that allows rapid, economic, and secure conduction of impulses along axons. The loss or lack of myelin resulting from an acquired or inherited disease may produce a delay or failure of conduction in affected fibers, with concomitant neurological dysfunction. In the human CNS, multiple sclerosis (MS) is the most common acquired demyelinating disease, affecting Ϸ2 million people worldwide (1). The leukodystrophies, induced by inherited enzyme deficiencies, also affect CNS white matter, resulting in abnormal formation, destruction, or turnover of myelin sheaths (2). Both acquired and inherited myelin disorders share a poor prognosis, leading to major disability in young people.Spontaneous remyelination can occur in the CNS and was first demonstrated by electron microscopy of lesions in the adult mammalian spinal cord (3). Remyelination results in the formation of short and thin myelinated internodes, but it enables the restoration of a sufficient conduction along axons and allows some functional recovery (4, 5). In demyelinating diseases such as MS, this regenerative process does occur and sometimes proceeds to completion (6), but it is less efficient than in experimental animal models (7). Improving repair processes can theoretically be achieved by either promoting endogenous repair mechanisms or providing an exogenous source of myelinating cells by transplantation (8-10). Clinical trials are expected to be carried out in the latter field soon.A major clinical issue of such trials is to assess and quantify myelin repair in vivo. To date, MRI is the reference test for diagnosing and monitoring the evolution of white-matter diseases (2, 11, 12). Unfortunatel...

No abstract

A method for in situ hybridization of digoxigenin-labeled cDNA and cRNA probes to myelin protein mRNA is described. This technique has dual advantages of high structural resolution and high sensitivity and avoids problems associated with handling of radioactive materials. Furthermore, it can be readily combined in double labeling with immunocytochemical protein detection. We have used this technique to detect and locate mRNA for myelin basic protein (MBP), proteolipid protein (PLP), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and myelin-associated glycoprotein (MAG) in oligodendrocytes of 7-day-old and adult rat brains. PLP and MAG mRNA were restricted to the perinuclear cytoplasm, whereas MBP and CNPase mRNA was additionally present in peripheral oligodendrocyte processes.

Summary: Children in America today are at an unacceptably high risk of developing neurodevelopmental disorders that affect the brain and nervous system including autism, attention deficit hyperactivity disorder, intellectual disabilities, and other learning and behavioral disabilities. These are complex disorders with multiple causes—genetic, social, and environmental. The contribution of toxic chemicals to these disorders can be prevented. Approach: Leading scientific and medical experts, along with children’s health advocates, came together in 2015 under the auspices of Project TENDR: Targeting Environmental Neuro-Developmental Risks to issue a call to action to reduce widespread exposures to chemicals that interfere with fetal and children’s brain development. Based on the available scientific evidence, the TENDR authors have identified prime examples of toxic chemicals and pollutants that increase children’s risks for neurodevelopmental disorders. These include chemicals that are used extensively in consumer products and that have become widespread in the environment. Some are chemicals to which children and pregnant women are regularly exposed, and they are detected in the bodies of virtually all Americans in national surveys conducted by the U.S. Centers for Disease Control and Prevention. The vast majority of chemicals in industrial and consumer products undergo almost no testing for developmental neurotoxicity or other health effects. Conclusion: Based on these findings, we assert that the current system in the United States for evaluating scientific evidence and making health-based decisions about environmental chemicals is fundamentally broken. To help reduce the unacceptably high prevalence of neurodevelopmental disorders in our children, we must eliminate or significantly reduce exposures to chemicals that contribute to these conditions. We must adopt a new framework for assessing chemicals that have the potential to disrupt brain development and prevent the use of those that may pose a risk. This consensus statement lays the foundation for developing recommendations to monitor, assess, and reduce exposures to neurotoxic chemicals. These measures are urgently needed if we are to protect healthy brain development so that current and future generations can reach their fullest potential.

Abbreviations used: CNP, 2¢, 3¢ cyclic nucleotide 3¢phosphodiesterase; EAE, experimental autoimmune encephalomyelitis; ErbB, epidermal growth factor receptor; Kv, voltage-gated potassium channels; MAG, myelin-associated glycoprotein; MS, multiple sclerosis; Nav, voltagegated sodium channels; NICD, Notch intracellular domain; NRG1, neuregulin-1; OPC, oligodendrocyte precursor cell; PLP, proteolipid protein; PSA-NCAM, poly-sialylated form of the neural cell adhesion molecule; TAG, transient axonal glycoprotein. AbstractIn multiple sclerosis, CNS demyelination is often followed by spontaneous repair, mostly achieved by adult oligodendrocyte precursor cells. Extent of this myelin repair differs, ranging from very low, limited to the plaque border, to extensive, with remyelination throughout the 'shadow plaques.' In addition to restoring neuronal connectivity, new myelin is neuroprotective. It reduces axonal loss and thus disability progression. Reciprocal communication between neurons and oligodendrocytes is essential for both myelin biogenesis and myelin repair. Hence, deciphering neuron-oligodendrocyte communication is not only important for understanding myelination per se, but also the pathophysiology that underlies demyelinating diseases and the development of innovative therapeutic strategies.

Radioactive choline was used to study the metabolism and movement of choline-containing phospholipids in peripheral nerve myelin of adult mice. Incorporation at various times after intraperitoneal injection was measured in serial segments of sciatic nerve as well as in myelin isolated from those segments. At no time (1 h to 35 days) could a proximal-distal difference in the extent of labeling be demonstrated. This finding suggests that incorporation of precursor choline phospholipids into nerve membranes is a local event, with little contribution from the neuronal perikaryon via axoplasmic transport.Autoradiographic investigations were undertaken to elucidate the pattern of movement of radioactive choline-labeled phospholipids, predominantly lecithin, into the myelin sheaths of the sciatic nerve. A sequence of autoradiographs was prepared from animals sacrificed between 20 min and 35 days after a microinjection of precursor directly into the nerve. Analysis of these autoradiograms revealed that labeling is initially concentrated in the Schwann cell cytoplasm. Later, the label moves first into the outer regions of the myelin sheaths and is eventually distributed evenly throughout the inner and outer layers of the sheath. At no time is there a build-up of label in the axon.The rate of uptake of precursor and subsequent redistribution of lecithin into the myelin were also examined in frog sciatic nerve (18~ Both uptake and redistribution processes were considerably slower in the cold-blooded animal.Various metabolic studies ( 15, 57) have shown that the sequence of events responsible for the dynamic turnover of membrane phospholipid (and protein) components includes synthesis in the endoplasmic reticulum and subsequent translocation to a variety of other membrane structures. Many recent reports have supported the contention that a turnover of myelin phospholipids occurs in the brain of adult animals (e.g. 9, 28, 29, 50). Miller and Dawson (34), employing subcellular fractionation of brain, have shown that most phospholipid biosynthesis including that of phosphatiylcholine takes place in "microsomes," with no activities detectable in isolated mitochondria or myelin. In addition, these authors (35) were unable to detect any component in brain cytoplasm which could

In the PNS, myelin basic protein (MBP) appears not to be essential for myelination, for in shiverer (shi) and mld mutant mice peripheral nerves, where MBP is not or only poorly expressed, myelination occurs normally. Only a few morphological abnormalities, i.e. reduction in axon calibre and myelin sheath thickness, and aberrant Schwann cell-axon contacts, have been reported. Here, we document a consistent difference between shi and wild type (wt) myelinated sciatic nerve fibres. The number of Schmidt-Lanterman incisures seen in longitudinally and transversely-sectioned sciatic nerves, or in teased fibres stained for the presence of F-actin, is dramatically increased in homozygous shi mice. With both methods, a twofold increase in Schmidt-Lanterman incisure number is seen in 15-day-old mice, the earliest time examined. The increase is slightly greater in nerve fibres from 30- and 90-day-old mice. The overproduction of Schmidt-Lanterman incisures in shi occurs in spite of the fact that the mean diameter of myelinated fibres in shi sciatic nerves is smaller than in wt sciatic nerves. These results lead us to suggest that the increase in Schmidt-Lanterman incisure density in shi compensates for a defect in Schwann cell-axon communication.

Abstract:The targeting of mRNAs to specific subcellular locations is believed to facilitate the rapid and selective incorporation of their protein products into complexes that may include membrane organelles. In oligodendrocytes, mRNAs that encode myelin basic protein (MBP) and select myelin-associated oligodendrocytic basic proteins (MOBPs) locate in myelin sheath assembly sites (MSAS). To identify additional mRNAs located in MSAS, we used a combination of subcellular fractionation and suppression subtractive hybridization. More than 50% of the 1,080 cDNAs that were analyzed were derived from MBP or MOBP mRNAs, confirming that the method selected mRNAs enriched in MSAS. Of 90 other cDNAs identified, most represent one or more mRNAs enriched in rat brain myelin. Five cDNAs, which encode known proteins, were characterized for mRNA size(s), enrichment in myelin, and tissue and developmental expression patterns. Two of these, peptidylarginine deiminase and ferritin heavy chain, have recognized roles in myelination. The corresponding mRNAs were of different sizes than the previously identified mRNA, and they had tissue and development expression patterns that were indistinguishable from those of MBP mRNA. Three other cDNAs recognize mRNAs whose proteins (SH3p13, KIF1A, and dynein light intermediate chain) are involved in membrane biogenesis. Although enriched in myelin, the tissue and developmental distribution patterns of these mRNAs differed from those of MBP mRNA. Six other cDNAs, which did not share significant sequence homology to known mRNAs, were also examined. The corresponding mRNAs were highly enriched in myelin, and four had tissue and developmental distribution patterns indistinguishable from those of MBP mRNA. These studies demonstrate that MSAS contain a diverse population of mRNAs, whose locally synthesized proteins are placed to contribute to myelin sheath assembly and maintenance. Characterization of these mRNAs and proteins will help provide a comprehensive picture of myelin sheath assembly.