High-resolution structural model of porcine P2 myelin membrane protein with associated fatty acid ligand: fact or artifact?

Myelin membrane is a biological complex of glial cells origin; it is composed of 25% (w/w) proteins and 75% lipids, and more than 300 proteins are associated with central nervous system myelin (for peripheral nervous system myelin, such data are lacking). Myelin plays an important role in maintaining propagation of nerve signals. To uncover the nature of propagation phenomena, it is essential to study biochemistry of myelin proteins and lipids, myelin composition, and myelin structure. Nearly all myelin proteins are like antigens, causing clinically well-defined devastating diseases; multiple sclerosis and Guillain-Barré syndrome are two of them. In this article, a high-resolution study (1.8 Å) of porcine myelin P2 protein is presented. Myelin was purified from porcine intradural spinal roots, which were stored at -80°C for 10 years before myelin and P2 protein were purified (spinal roots were a gift of Prof. Kunio Kitamura, Saitama Medical School). The three-dimensional structural analysis uncovered embedded 18-carbons-long fatty acid. Some speculative interpretation is presented, to uncover how this ligand of fatty acid may form cholesterol ester and stabilize the myelin structure or form simple raft microdomain. Protein crystallography indicates that the ligand may be 18-carbons-long fatty acid. This is unlike previous work with mass spectrometry, in which three ligands were determined. In other protein crystallography-based studies of P2 (bovine), an oleic fatty acid was suggested, but, for recombinant (human) protein, palmitic acid was found. There is no fatty acid ligand in equine P2 protein.

Mifepristone (RU 38486) influences expression of glycoprotein Po and morphological parameters at the level of rat sciatic nerve: in vivo observations.

The observations here reported indicate that, in vivo, the expression of an important protein of peripheral myelin, the glycoprotein Po, is influenced by mifespristone (RU 38486), that is, an antagonist of progesterone (PR) and glucocorticoid (GR) receptor. In our experimental model, male rats have been treated at the first day of life with this antagonist and after repeated treatments, we have analyzed in the sciatic nerve of 20- (20d) and 30-day-old rats (30d) the mRNA and protein levels of Po. Moreover, expression of Po has also been analyzed in the sciatic nerve of animals treated during the first 30 days of postnatal life and then sacrificed at 90th day of life (90d). The results obtained have indicated that both mRNA and protein levels of Po decrease at 20d. Apparently, these effects seem to be transient because no changes are evident at the other two times of analysis. As shown by morphometric analysis, the treatment with RU 38486 is also able to induce morphological changes at the level of sciatic nerve. However, at variance to what is expected by an alteration of an important component of the myelin membranes like Po, no changes are evident at the level of the myelin compartment. On the contrary, a significant reduction of axon diameter in parallel to an increase in neurofilament (NF) density occurs since 30d. In conclusion, the present data seem to suggest that progestin and/or glucocorticoid signals are not only involved in the control of myelin compartment but also on the axon maintenance.

Characterization of myelination in the developing zebrafish.

Myelination, the process by which glial cells ensheath and electrically insulate axons, has been investigated intensely. Nevertheless, knowledge of how myelination is regulated or how myelinating cells communicate with neurons is still incomplete. As a prelude to genetic analyses of these processes, we have identified zebrafish orthologues of genes encoding major myelin proteins and have characterized myelination in the larval zebrafish. Expression of genes corresponding to proteolipid protein (PLP/DM20), myelin protein zero (P0), and myelin basic protein (MBP) is detected at 2 days postfertilization (dpf), first in the ventral hindbrain, close to the midline. During the next 8 days, expression spreads rostrally to the midbrain and optic nerve, and caudally to the spinal cord. DM20 is expressed in the CNS only, while MBP transcripts are detected both in the CNS and in Schwann cells of the lateral line, cranial nerves, and spinal motor nerves. Unlike its closest homologue, trout IP1, zebrafish P0 transcripts were restricted to the CNS. Ultrastructurally, the expression of myelin genes correlated well with myelination, although myelination showed a temporal lag. Myelinated axons were first detected at 4 dpf in the ventral hindbrain, where they were loosely wrapped by processes of glia cells. By 7 dpf, bundles of heavily myelinated axons were observed in the same region. Axons in the lateral line and optic nerves were also surrounded by compact myelin. Conservation in gene expression patterns and the early appearance of myelinated axons, support using the zebrafish to dissect the process of myelination by a genetic approach.

Clinically distinct codon 69 mutations in major myelin protein zero in demyelinating neuropathies.

Mutations in the major peripheral myelin protein zero (P0) gene on chromosome 1q21-q23 have been found with the hereditary demyelinating polyneuropathy Charcot-Marie-Tooth type 1B. Here, we describe 2 patients with distinct neurological characteristics, carrying different substitutions at the same codon--Arg69His and Arg69Cys. The patients were heterozygous for the mutation, which in both appeared to be de novo. Histological examination of sural nerve biopsy specimens revealed defective myelin as well as marked differences, confirming the importance of P0 in the compaction of myelin.