Postnatal delay of myelin formation in brains from Down syndrome infants and children.

Myelination up to the age of 6 years was studied in two groups consisting of 129 Down syndrome (DS) 17/129 fetuses and 112/129 postnatal) and 73 non-DS cases (10/73 fetuses and 63/73 postnatal). In both groups studied a similar number of congenital heart disease (CHD), gastrointestinal (GI) malformations and infections were diagnosed. Paraffin or celloidin brain sections were stained with Klüver-Barrera, Heidenhain or Loyez method. The myelination was found to be delayed in 29/129 (22.5%) DS and only in 5/73 (6.8%) non-DS cases. Myelination in fetuses and newborns in the DS and non-DS groups was not delayed. In DS the myelination was delayed between ages 2 months-6 years (17/29; 58.6%) up to 12 months, and 12/29 (41.4%) aged 2-6 years, while in non-DS aged 2-6 months only. The myelination delay affected tracts with late beginning and slow cycle of myelination, mainly the associated and intercortical fibers of the fronto-temporal lobes. In 3/7 of DS cases (ages 3, 4, 6 years) less advanced myelination of U fibers was noted. In both groups the myelination delay seemed also to depend on the systemic diseases which affected the subjects during the time of myelination. In DS where CHD was present the myelination delay was found in 14/29 (48.2%) in contrast to 3/5 (60%) non-DS. Also, in 28/129 (23%) DS cases dates regarding the developmental milestones were available and some correlation was found between developmental and myelination delay.(ABSTRACT TRUNCATED AT 250 WORDS)

Developmental features of the neonatal brain: MR imaging. Part I. Gray-white matter differentiation and myelination.

To establish the normal appearance of the neonatal brain, 51 neonates, 29-42 weeks postconception, underwent magnetic resonance (MR) imaging with a 0.6-T magnet in a prospective study. T1-weighted images were used to devise stages for the appearance of gray-white matter differentiation and extent of myelination. The results show that from 29 to 42 weeks postconception, changes in gray-white matter differentiation and myelination follow the stages in an orderly and predictable fashion. Changes in white matter intensity appear related to progressive decrease in brain water content. Myelination progresses cephalad from the brain stem at 29 weeks to reach the centrum semiovale by 42 weeks. Delayed myelination, defined as the absence of myelin in the corona radiata by 37 weeks, was seen in nine infants with complicated perinatal courses. Awareness of these developmental features should help to minimize misinterpretation of normal changes in the neonatal brain and lead to earlier detection of pathologic conditions, both with MR imaging and computed tomography.

In vivo phosphorylation of myelin basic proteins: age-related differences in 32P incorporation.

Myelin basic proteins (MBPs) are phosphoproteins of central and peripheral nervous system myelin. We studied the phosphorylation of mouse MBPs in vivo at three different stages of development (12, 30, and 50 days) and found age-related differences in the incorporation of 32P into MBPs. At all ages studied, significant amounts of 32P were found in the MBPs as early as 1 min after intracranial injection of isotope. Incorporation of radioactive phosphate into MBPs proceeded rapidly and the resultant specific radioactivity (SA) of 32P-labeled MBPs appeared to be related to the SA of the acid-soluble phosphate pool of myelin. Changes in the SA of the myelin acid-soluble phosphate pool were observed in a 30 min time course of labeling in vivo in 50-day mice. Coincident changes were observed in the SA of the MBPs. Similar but less pronounced changes were seen in the SA of the polyphosphoinositides (PPIs) indicating that the turnover of the PPI phosphate groups is slower than the MBP phosphates or that the PPI phosphates are drawn from additional or different pools than the MBP phosphates. The phosphorylation of MBPs in developmentally related myelin fractions is investigated in a comparison paper (J. B. Ulmer and P. E. Braun (1986) Dev. Biol. 117, 502-510).

In vivo phosphorylation of myelin basic proteins: single and double isotope incorporation in developmentally related myelin fractions.

The phosphorylation of myelin basic proteins (MBPs) was studied in developing mouse brain. Based on our previous work we postulated that phosphorylation of MBPs takes place prior to their appearance in the myelin compartment as well as within the myelin sheath. To further test this hypothesis we utilized a subfractionation protocol that yields brain fractions enriched in myelin membranes of differing developmental stages. Incorporation of radioactive phosphate into MBPs was studied in each of the subcellular fractions. After 5- and 15-min incubations of isotope in vivo the highest specific radioactivities (SAs) of MBPs were found in the least mature myelin fractions. Incorporation of 32P in MBPs was greater into serine residues than threonine residues in all of the subcellular fractions studied. The relative turnover of MBP phosphates was studied in each of the subcellular myelin fractions using a time-staggered, double isotope methodology. The most rapid equilibration of MBP phosphates with the trichloroacetic acid (TCA)-soluble phosphate pool occurred in the most mature myelin fractions indicating that the highest turnover of MBP phosphates occurs in the most mature myelin fractions. The SAs and turnover rates of each of the four commonly observed mouse MBPs (14, 17, 18.5, and 21.5 kDa) were similar in any particular subfraction demonstrating that the MBP phosphotransferase system(s) acts on each of the MBPs in a similar manner.

Effect of halothane on brain 2',3'-cyclic nucleotide 3'-phosphodiesterase during neurodevelopment in the rat.

Chronic exposure to anesthetic concentrations of halothane during the prenatal and early postnatal periods inhibits the incorporation of the leucine into myelin subcellular fractions in the rat. The enzyme 2',3' - cyclic nucleotide 3' - phosphodiesterase (CNPase) has been widely used as a myelin marker. To determine the effect of halothane on the developmental profile of CNPase, two groups of pregnant Sprague Dawley rats were exposed to 500 p.p.m. or 250 p.p.m. halothane, eight hours per day, five days per week from the third day after conception through postnatal day ten. Control animals were exposed to air alone. CNPase activity was significantly decreased by 500 p.p.m. halothane (34%) and by 250 p.p.m. halothane (29%) at postnatal day 17. Brain and body weights in both halothane treated groups were also less than control animals throughout the measurement period. The data indicates that chronic pre- and postnatal halothane exposure at low levels delays myelination in the rat.

MRI of normal brain maturation.

The unprecedented gray/white differentiation obtained with magnetic resonance imaging (MRI) has created a unique opportunity to trace the normal process of myelination. Fifty-nine children referred for evaluation of a nonneurologic problem or a nonspecific neurologic complaint were studied with MRI using spin-echo technique. Children ranged in age from term (40 weeks intrauterine) to 16 years. Scans were reviewed for quantitative and qualitative changes with age. T1 and T2 relaxation times were measured for 13 regions of interest in 37 children. With increasing age a sharp decrease in both T1 and T2 values, most pronounced during the first year of life, was seen. The prolonged relaxation times in the newborn infant correspond to the known high water content of the neonatal brain; the subsequent decline corresponds to the decrease in water content and increase in myelination observed in autopsy studies of infants. Qualitative changes in the MRI appearance of the brain with age using a spin-echo sequence (2 sec repetition time) demonstrated that the process of myelination was most rapid during the first 2-3 years of life. Myelination appeared to occur earliest in the posterior fossa, with the middle cerebellar peduncle identifiable at only 3 months. By the age of 1 year, all major white matter tracts including the corpus callosum, subcortical white matter, and the internal capsule were well defined. However, due to subtle changes in appearance, the refined configuration of the adult brain was not attained until early adolescence.

Immunocytochemical demonstration of glial-neuronal interactions and myelinogenesis in subcultures of rat brain cells.

Subcultures have been established from primary rat brain cell cultures and have been characterised with a range of cell-specific immunocytochemical markers. The subcultures are mainly composed of fibrous astrocytes, oligodendrocytes and neurones. The cells do not divide to any great extent giving a system where it is possible to follow culture development at the cellular level for a number of weeks. During this time oligodendrocytes colonise subpopulations of neurones, differentiate further showing the presence of myelin basic protein and elaborate myelin-like membrane; the fibrous astrocytes remain scattered uniformly throughout the cultures. Radially oriented processes emerge from the oligodendrocyte-neurone aggregates which subsequently coalesce to form fascicles that link the clusters of cells together. These fascicles react with antibodies for both neurofilament protein and myelin basic protein. The subcultures provide a straightforward system that is composed of cells derived entirely from the CNS, is free from mitotic inhibitors and yet retains a sufficiently low cell density to allow immunocytochemical identification of the cell types present. The subcultures should be useful for the study of trophic interactions between oligodendrocytes and neurones as well as the early events associated with myelinogenesis.

Peripheral nerves in shiverer----dystrophic mouse chimeras: evidence that a non-Schwann cell component is required for axon ensheathment in vivo.

Peripheral nerves in dystrophic mice express multiple axon ensheathment abnormalities. If an intrinsic deficiency expressed within the Schwann cells themselves were to account for this neuropathy, then such cells, existing in a chimera preparation, would be expected to express the same ensheathment abnormalities, whereas a coexisting population of non-dystrophic Schwann cells should not be similarly affected. The genotype of myelinated Schwann cells in shiverer----dystrophic chimera was established with immunocytochemical techniques. Shiverer myelin lacks the P1 component of myelin basic protein (MBP), whereas dystrophic myelin appears to contain normal levels of MBP. No correlation between the ensheathment characteristics of the chimera spinal roots and the genotype of the local Schwann cell population was found; both dystrophic Schwann cell populations expressing normalized ensheathment characteristics and shiverer Schwann cells failing to respond to the local presence of naked axons were observed. These results require that a defective extra Schwann cell component is involved in the pathogenesis of the dystrophic neuropathy. Moreover, the normal realization of that component appears to be a necessary prerequisite for shiverer Schwann cells to achieve full ensheathment competence. Although a definitive identification of the cell type(s) that expresses the dy gene locus has not been achieved in this chimera preparation, the observations are consistent with defective endoneurial fibroblast function.

Impaired Myelination in rats given excess vitamin A postnatally.

The effect of postnatal administration of 1000 I.U. of Vitamin A on 4th, 6th, 8th, and 10th day of age to rat pups has been studied on brain myelin lipid and sulphatide synthesis from Na235SO4. Administration of vitamin A reduced brain weight, free cholesterol, phosphatidal ethanolamine and the synthesis of myelin sulphatides from Na235SO4.

Changing relation between onset of myelination and axon diameter range in developing feline white matter.

The size spectra of unmyelinated, ensheathed and initially myelinating CNS axons were examined by electron microscopy in the spinal cord ventral funiculus and the corpus callosum of the cat during development. The first myelin sheaths appeared 4 weeks before and 3 weeks after birth in the spinal and callosal areas, respectively. De novo myelination had largely ceased by 4 months in the ventral funiculus and by 7 months in the corpus callosum. The result show that the diameter ranges, within which spinal and callosal axons undergo primary ensheathment and initial myelination, are markedly different if similar levels of myelination are compared. In both areas, these diameter ranges shift towards smaller sizes with development. However, spinal and callosal axons, which undergo primary ensheathment and initial myelination simultaneously, present comparable diameter ranges. The findings support the view that other factors than the absolute physical size of the axon trigger initiation of CNS myelination. In this respect the developmental stage of the animal appears to play an important role.

The ultrastructure of sheath cells in developing rat vomeronasal nerve.

Maturation of the vomeronasal nerve was studied in fetal, newborn and 3 months old rats. Early in development, each nerve consists of large numbers of bare axons with clusters of polygonal sheath cells lying at the periphery. The latter insinuate themselves between the axons which they segregate into bundles. The sheath cells and their processes which first delineate axon bundles from one another form a network in the interstices of which lie the emergent axon bundles. Each sheath cell is not confined to the sleeve around a single bundle. Its perikaryon and processes commonly contribute to the septa between several adjacent bundles. Eventually, each bundle comes to be surrounded by its own proper sheath which consists of processes of more than one Schwann cell. These developmental trends, of a progressive increase in the number both of axons per bundle and of Schwann cells associated with each bundle, are the reverse of those found in the PNS generally, where bundle size decreases and axon size increases with maturation. As individual bundles separate from one another, interfascicular collagen appears between them and each comes to be surrounded by a basal lamina. Separation is rarely complete, however; even at the mature stage, processes are exchanged between adjacent sheaths at one or more points on their circumferences. Schwann cell processes surrounding individual bundles become increasingly complex with maturation. Where adjoining processes meet, they commonly branch profusely and interdigitate intricately, forming stacks of closely apposed layered processes. In other areas, the branches are covered by basal lamina and bound intricate labyrinths which commonly extend deeply into the bundle and contain collagen fibrils.

Effects of alpha-methylphenylalanine plus phenylalanine treatment during development on myelin in rat brain.

The effects of hyperphenylalaninemia induced by treatment with alpha-methylphenylalanine (MPA) plus phenylalanine (PHE) on body and brain weight, on myelin and synaptosome formation, and on the lipids and fatty acids of myelin were studied in rats. The administraton of MPA (2.4 mumol/g body wt) plus PHE (2.6 mumol/g body wt) for 25 and 35 days beginning on the fifth postnatal day did not affect brain development. On doubling the dosage of PHE, body and brain weights and myelin yields were significantly lowered. The lipid composition of myelin from the brains of treated animals was largely unaffected; however, the concentration of sulfatides ws significantly reduced. Unsaturated fatty acid levels in myelin from hyperphenylalaninemic rat brains were reduced while long-chain fatty acids were unaffected. We conclude that as in hyperphenylalaninemia induced by other methods, MPA + PHE treatment impairs body and brain growth, reduces myelin formation, and causes inhibition of fatty acid desaturation in the brain.

Myelin basic protein demonstrated immunocytochemically in oligodendroglia prior to myelin sheath formation.

A specific antibody to myelin basic protein has been used to localize the protein in developing rat oligodendroglia and myelin. Basic protein is found in the oligodendroglial cytoplasm of anterior commissures of 5- and 7-day old rats before the beginning of myelination. Staining of basic protein in oligodendroglia increases, becoming most intense during early myelination; it decreases during rapid myelination. Staining intensity of oligodendroglia is dependent upon age, brain region, and nervous tract studied. In myelin, reaction of basic protein with antibody decreases when large compact sheaths are present, unless tissue sections are first treated with alcohol.