Down Syndrome Developmental Brain Transcriptome Reveals Defective Oligodendrocyte Differentiation and Myelination.

Trisomy 21, or Down syndrome (DS), is the most common genetic cause of developmental delay and intellectual disability. To gain insight into the underlying molecular and cellular pathogenesis, we conducted a multi-region transcriptome analysis of DS and euploid control brains spanning from mid-fetal development to adulthood. We found genome-wide alterations in the expression of a large number of genes, many of which exhibited temporal and spatial specificity and were associated with distinct biological processes. In particular, we uncovered co-dysregulation of genes associated with oligodendrocyte differentiation and myelination that were validated via cross-species comparison to Ts65Dn trisomy mice. Furthermore, we show that hypomyelination present in Ts65Dn mice is in part due to cell-autonomous effects of trisomy on oligodendrocyte differentiation and results in slower neocortical action potential transmission. Together, these results identify defects in white matter development and function in DS, and they provide a transcriptional framework for further investigating DS neuropathogenesis.

Age-related changes in the morphology of cerebral capillaries do not correlate with cognitive decline.

The effects of age on cerebral capillaries have been examined in area 46 of the prefrontal cortices of sixteen rhesus monkeys, ranging in age from 5 to 35 years. Fourteen of the monkeys had been behaviorally tested prior to their brains being prepared for electron microscopic examination. It was found that whereas the thickness of the outer basal lamina adjacent to the glial limiting membrane increased with age and showed increasing numbers of splits, the inner basal lamina between endothelial cells and pericytes did not become thicker with age, and did not show splitting. There were also no age-related changes in the extent of the coverage of endothelial cells by pericytes and no change in the frequency of mitochondria in endothelial cells. The factors that did change with age, namely, the thickness of the outer basal lamina and the increased numbers of splits in this lamina showed no correlations with the cognitive status of the monkeys, suggesting that thickening of the outer basal lamina does not contribute to cognitive decline.

How the primate fornix is affected by age.

The effects of age on nerve fibers and neuroglial cells in the fornix were examined in 25 rhesus monkeys between 4 and 33 years of age. There is no age-related change in the cross-sectional area of the fornix, but there is a significant loss of myelinated nerve fibers. The loss of myelinated nerve fibers is accompanied by a significant increase in the numbers of nerve fibers that show degeneration of their axons and alterations in myelin sheaths. Aging also brings about an increase in the frequency of profiles of paranodes, indicating that some of the nerve fibers are being remyelinated. Aging also affects neuroglial cells. Each type shows inclusions in their perikarya, and in the case of astrocytes and microglial cells some of these inclusions are phagocytosed myelin. Numbers of astrocytes and microglial cells do not appear to increase with age, but there is a 20% increase in oligodendrocytes. When correlations with cognitive impairments displayed by individual monkeys are examined, the decreased packing density of nerve fibers and the increasing frequency of nerve fibers with degenerating axons and of nerve fibers with altered myelin sheaths all correlate with increasing cognitive impairment. It is suggested that these correlations result from some disconnection of the hippocampus from the thalamus, septal nuclei, and medial frontal cortex and from reductions in the conduction velocity brought about by the shorter internodal lengths of remyelinated nerve fibers in the fornix.

The neuroglial population in the primary visual cortex of the aging rhesus monkey.

The effects of age on neuroglial cells have been examined in the primary visual cortices of rhesus monkeys that had been behaviorally tested. The assessment of changes in the neuroglial populations was made on the basis of the frequency of occurrence of profiles of neuroglial cells in semithick sections of osmicated tissue stained with toluidine blue. No changes were found in the numbers of astrocytes and microglial cells with age, but the numbers of oligodendrocytes increased by about 50%. The myelinated nerve bundles at the level of layer 4 were also examined by electron microscopy to assess the effects of age on the nerve fibers. The numbers of nerve fiber profiles showing age-related alterations in their myelin sheaths increase with age. There was also an age-related increase in the frequency of profiles of nerve fibers sectioned through paranodes, indicating that shorter lengths of myelin are being produced by remyelination. These changes in sheaths both correlate significantly with the frequency of oligodendrocyte profiles, suggesting that with age additional oligodendrocytes are required to remyelinate nerve fibers whose sheaths have broken down, probably by death of the original parent oligodendroglial cell. Also the most cognitively impaired monkeys had the greatest numbers of oligodendrocytes, but this is probably a secondary correlation, reflecting the fact that altered myelin slows down the rate of conduction along nerve fibers, which leads to cognitive decline.

Oligodendrocytes, their progenitors and other neuroglial cells in the aging primate cerebral cortex.

In a previous study it was found that with age there is an increase in the frequency of paranodal profiles of myelinated nerve fibers in the cerebral cortex of monkeys. This indicates that there is an increase in the number of internodal myelin segments, and raises the question of whether additional oligodendrocytes are necessary to generate the increased numbers of internodal myelin segments. The present study shows that in layer 4C beta of monkey primary visual cortex there is an age-related increase in the number of oligodendrocytes. When young (4-10 years of age) and old (25-35 years of age) monkeys are compared, the increase is found to be approximately 50%, and it begins in middle age (12-19 years old). It is also shown that although there is no increase in the population of astrocytes in layer 4C beta with age, there appears to be a slight increase in the frequency of microglial cells. As their numbers increase, oligodendrocytes in pairs, rows and groups become more common, which suggests that additional oligodendrocytes are being generated by cell division. Since there is little evidence that mature oligodendrocytes can divide, it is probable that the new oligodendrocytes are generated from progenitor cells which, as many studies have shown, can be labeled by antibodies to NG2, a chondroitin sulfate proteoglycan. By comparing the appearance of these NG2-labeled cells with cells encountered in thin sections of normally prepared tissue, it is shown that the NG2-positive cells have the features of neuroglial cells that were previously described as beta astrocytes.

Is there remyelination during aging of the primate central nervous system?

The effect of aging on myelin sheaths in the rhesus monkey was studied in the vertical bundles of nerve fibers that traverse monkey cerebral cortex in primary visual area 17 and prefrontal area 46. As shown previously, with age the internodes of many of these myelin sheaths show structural changes, the most common of which is an accumulation of electron-dense cytoplasm within some sheaths, a change which is considered to indicate that breakdown of myelin is taking place. Supporting the suggestion that myelin is breaking down with age, astrocytes in the cortices of old monkeys contain phagocytosed myelin and some of the inclusion bodies in astrocytes label with antibodies to myelin basic protein. There is also evidence that remyelination is taking place. Thus, we have found an increase in the frequency of profiles of paranodes when transverse sections of the nerve fibers are examined. The increase in paranodal frequency with age is 57% in area 17 and 90% in area 46. This increase cannot all be attributed to lengthening of paranodes with age, because in area 17 the 11% increase in mean paranodal length with age is insufficient to account for an age-related increase in paranodal profile frequency. Consequently, there must be an increase in the number of internodal lengths of myelin with age, as would occur if shorter lengths of myelin are produced by remyelination. In support of the proposal that remyelination is occurring, short internodal lengths of myelin have been found in the nerve bundles passing through the cortices of old monkeys and inappropriately thin sheaths occur around some axons. Both of these features are generally considered to be the hallmarks of remyelination. Consequently, it is proposed that in the aging cerebral cortex of the monkey there is some breakdown of internodes of myelin with subsequent remyelination that leads to the formation of some new and shorter internodal lengths of myelin.

Aging and the myelinated fibers in prefrontal cortex and corpus callosum of the monkey.

In the rhesus monkey, the myelin sheaths of nerve fibers in area 46 of prefrontal cortex and in splenium of the corpus callosum show age-related alterations in their structure. The alterations are of four basic types. Most common is splitting of the dense line of myelin sheaths to accommodate electron dense cytoplasm derived from the oligodendroglia. Less common are splits of the intraperiod line to form balloons or blisters that appear to contain fluid, the occurrence of sheaths with redundant myelin, and thick sheaths that are almost completely split so that one set of compact lamellae is surrounded by another set. But despite these alterations in the sheaths, few nerve fibers show axonal degeneration. To quantify the frequency of the age-related alterations in myelin, transversely sectioned nerve fibers from the splenium of the corpus callosum and from the vertical bundles of nerve fibers within area 46 were examined in electron photomicrographs. The material was taken from 19 monkeys, ranging between 5 and 35 years of age. It was found that the frequency of alterations in myelin sheaths from both locations correlates significantly with age. In area 46, the age-related alterations also significantly correlate (P < 0.001) with an overall assessment of impairment in cognition, i.e., the cognitive impairment index, displayed by individual monkeys. The correlation is also significant when only the old monkeys are considered as a group. A similar result was obtained previously in our examination of the effects of age on the myelin sheaths of nerve fibers in primary visual cortex (Peters et al. [2000] J Comp Neurol. 419:364-376). However, in the corpus callosum the myelin alterations correlate significantly with only one component of the cognitive impairment index, namely the delayed nonmatching to sample task with a 2-minute delay. It is proposed that age-related myelin alterations are ubiquitous and that the correlations between their frequency and impairments in cognition occur because the conduction velocity along the affected nerve fibers is reduced, so that the normal timing sequences within neuronal circuits break down.

The effects of age on the cells in layer 1 of primate cerebral cortex.

Although there is significant thinning of layer 1 with age in both occipital area 17 and prefrontal area 46 of the rhesus monkey, there are no significant age-related changes in the numbers of neurons, astrocytes, or microglia and oligodendrocytes in this layer. A few profiles of degenerating neurons have been encountered in old monkeys, but they are uncommon. Some astrocytes undergo hypertrophy with age, as evidenced by the increased thickness of the glial limiting membrane, and throughout layer 1 the amount of filaments in the cytoplasm of both their cell bodies and processes increases. The astrocytes also come to contain phagocytic material in the old monkeys, as do the microglial cells. We have previously shown that in both areas 17 and 46 there is an age-related loss of synapses from layer 1 and a concomitant loss of dendritic branches from the apical tufts of pyramidal cells from layer 1. These may be the sources of the material phagocytosed by the astrocytes and microglial cells.