Human Brain White Matter Research Articles

White Matter Fiber Tracking Computation Based on Diffusion Tensor Imaging for Clinical Applications

Fiber tracking allows the in vivo reconstruction of human brain white matter fiber trajectories based on magnetic resonance diffusion tensor imaging (MR-DTI), but its application in the clinical routine is still in its infancy. In this study, we present a new software for fiber tracking, developed on top of a general-purpose DICOM (digital imaging and communications in medicine) framework, which can be easily integrated into existing picture archiving and communication system (PACS) of radiological institutions. Images combining anatomical information and the localization of different fiber tract trajectories can be encoded and exported in DICOM and Analyze formats, which are valuable resources in the clinical applications of this method. Fiber tracking was implemented based on existing line propagation algorithms, but it includes a heuristic for fiber crossings in the case of disk-shaped diffusion tensors. We successfully performed fiber tracking on MR-DTI data sets from 26 patients with different types of brain lesions affecting the corticospinal tracts. In all cases, the trajectories of the central spinal tract (pyramidal tract) were reconstructed and could be applied at the planning phase of the surgery as well as in intraoperative neuronavigation.

Mapping the intracellular fraction of water by varying the gradient pulse length in q-space diffusion MRI

Finite gradient pulse lengths are traditionally considered a nuisance in q-space diffusion NMR and MRI, since the simple Fourier relation between the acquired signal and the displacement probability is invalidated. Increasing the value of the pulse length leads to an apparently smaller value of the estimated compartment size. We propose that q-space data at different gradient pulse lengths, but with the same effective diffusion time, can be used to identify and quantify components with free or restricted diffusion from multiexponential echo decay curves obtained on cellular systems. The method is demonstrated with experiments on excised human brain white matter and a series of model systems with well-defined free, restricted, and combined free and restricted diffusion behavior. Time-resolved diffusion MRI experiments are used to map the spatial distribution of the intracellular fraction in a yeast cell suspension during sedimentation, and observe the disappearance of this fraction after a heat treatment.

Proteomic studies on the white matter of human brain☆

Limited information on the protein expression profiles of the different components of mammalian brain is available to date. In the present study, proteomic analysis was performed on 32 white matter samples obtained from 8 different regions of brains of four post mortem cases. Proteins were separated by 2D gel electrophoresis and identified by mass spectrometry. Most of the protein spots (98%) are reproducibly present in all the samples analyzed. A total of 64 different proteins were identified and divided into seven functional groups. These include metabolic proteins (33%), structural proteins (9%), proteins involved in signal transduction (9%), blood proteins (8%), stress related proteins (23%), and proteins involved in the ubiquitin mediated proteolysis (6%). This protein database obtained from the white matter of human brain contributes to deepen our knowledge on the molecular mechanisms that control several pathologies affecting this key component of the brain.

White matter fiber tracts of the human brain: Three-dimensional mapping at microscopic resolution, topography and intersubject variability

The position and extent of individual fiber tracts within the white matter of human brains can be identified in vivo using diffusion tensor imaging (DTI) and fiber tracking methods. Previous to this study, however, the lack of three-dimensional (3-D) probability maps precluded comparing the anatomical precision of MRI studies with microscopically defined fiber tracts in human postmortem brains. The present study provides 3-D registered maps of the topography, course and intersubject variability of major fiber tracts, which were identified at microscopic resolution. The analyzed tracts include the corticospinal tract, optic and acoustic radiations, fornix, cingulum, corpus callosum, superior longitudinal, superior and inferior occipito-frontal and uncinate fascicles; sources and targets of fiber tracts include the lateral and medial geniculate nuclei and mamillary bodies. Tracts and nuclei were identified in serial myelin-stained histological sections of ten postmortem brains. The sections were 3-D reconstructed and registered to a standardized stereotaxic space of an in vivo MR reference brain by means of linear and non-linear, elastic transformations. The individual fiber tracts and nuclei were superimposed in the reference space, and probability maps were generated as a quantitative measure of intersubject variability for each voxel of the stereotaxic space. This study presents the first stereotaxic atlas of the course, location and extent of fiber tracts and related nuclei based on microscopically defined localization and topographic data taken at multiple levels on each of the three orthogonal planes. The maps are useful for evaluating and identifying fiber bundles in DTI, for localizing subcortical lesions visible in anatomical MR images and for studying neuronal connectivity.

A three-dimensional fractal analysis method for quantifying white matter structure in human brain

Fractal dimension (FD) is increasingly used to quantify complexity of brain structures. Previous research that analyzed FD of human brain mainly focused on two-dimensional measurements. In this study, we developed a three-dimensional (3D) box-counting method to measure FD of human brain white matter (WM) interior structure, WM surface and WM general structure simultaneously. This method, which firstly incorporates a shape descriptor (3D skeleton) representing interior structure and combines the three features, provides a more comprehensive characterization of WM structure. WM FD of different brain segments was computed to test robustness of the method. FDs of fractal phantoms were computed to test the accuracy of the method. The consistency of the computed and theoretical FD values suggests that our method is accurate in measuring FDs of fractals. Statistical analysis was performed to examine sensitivity of the method in detecting WM structure differences in a number of young and old subjects. FD values of the WM skeleton and surface were significantly greater in young than old individuals, indicating more complex WM structures in young people. These results suggest that our method is accurate in quantifying three-dimensional brain WM structures and sensitive in detecting age-related degeneration of the structures.

Expression of myelin basic protein (MBP) epitopes in human non-neural cells revealed by two anti-MBP IgM monoclonal antibodies.

Two monoclonal antibodies (1H6.2 and 45.30) were raised against MBP purified from human brain under experimental conditions that allowed MBP to retain binding to surrounding myelin lipids (human lipid-bound MBP (hLB-MBP)). 1H6.2 and 45.30 MoAbs were selected on the basis of their different binding properties to: hLB-MBP, human lipid-free-MBP (hLF-MBP) and bovine lipid-free-MBP (bLF-MBP). Although the isotype of both MoAbs was IgM, their specificity, as tested in ELISA assays against chemical haptens and unrelated protein antigens, was restricted to MBP. 1H6.2 and 45.30 MoAbs stained MBP from human brain white matter tissue extracts, as well as bLF-MBP, in Western blot assays. Both MoAbs stained oligodendrocytes and myelin in immunohistochemical analysis of white matter from human brain. Tissue sections from human peripheral nerves were labelled by 1H6.2 only, however, demonstrating that the MoAbs recognize two different epitopes. Epitopes recognized by 1H6.2 and 45.30 MoAbs were also expressed by a wide array of human non-neural cells of either normal or pathological origin, as evidenced by cytofluorimetric assays. In particular, MBP epitopes (MEs) were expressed by lymphoid cells as well as by cells which play a pivotal role in immune homeostasis and in the immune response, such as thymic epithelial cells and professional antigen-presenting cells. Both MoAbs were efficiently internalized by cells from a human B cell line, suggesting trafficking of MEs along the endocytic pathways. These findings support hypotheses regarding the role of MEs expressed by non-neural cells in establishing self-tolerance and/or in triggering the immune response against MBP antigen.

Differentiation of glucose transport in human brain gray and white matter.

Localized 1H nuclear magnetic resonance spectroscopy has been applied to determine human brain gray matter and white matter glucose transport kinetics by measuring the steady-state glucose concentration under normoglycemia and two levels of hyperglycemia. Nuclear magnetic resonance spectroscopic measurements were simultaneously performed on three 12-mL volumes, containing predominantly gray or white matter. The exact volume compositions were determined from quantitative T1 relaxation magnetic resonance images. The absolute brain glucose concentration as a function of the plasma glucose level was fitted with two kinetic transport models, based on standard (irreversible) or reversible Michaelis-Menten kinetics. The steady-state brain glucose levels were similar for cerebral gray and white matter, although the white matter levels were consistently 15% to 20% higher. The ratio of the maximum glucose transport rate, V(max), to the cerebral metabolic utilization rate of glucose, CMR(Glc), was 3.2 +/- 0.10 and 3.9 +/- 0.15 for gray matter and white matter using the standard transport model and 1.8 +/- 0.10 and 2.2 +/- 0.12 for gray matter and white matter using the reversible transport model. The Michaelis-Menten constant K(m) was 6.2 +/- 0.85 and 7.3 +/- 1.1 mmol/L for gray matter and white matter in the standard model and 1.1 +/- 0.66 and 1.7 +/- 0.88 mmol/L in the reversible model. Taking into account the threefold lower rate of CMR(Glc) in white matter, this finding suggests that blood--brain barrier glucose transport activity is lower by a similar amount in white matter. The regulation of glucose transport activity at the blood--brain barrier may be an important mechanism for maintaining glucose homeostasis throughout the cerebral cortex.

A variant very low density lipoprotein receptor lacking 84 base pairs of O-linked sugar domain in the human brain myelin

The very low density lipoprotein receptor (VLDLR) was considered to specifically bind to VLDL rich apolipoprotein E (apoE). However, its distribution and functions in vivo have yet to be elucidated. In human and rat VLDLR, a variant form lacking 84 base pairs (bp) in O-linked sugar domain was noted but its significance was not initially understood. This study shows that the variant form of VLDLR coexists with full-length VLDLR in majority of tissues but is a major component in the white matter of human brain. The tissue distribution of a variant VLDLR was detected in myelin as well as in other tissues except for the liver with immunohistochemistry using a monoclonal antibody. This variant VLDLR is proposed to be functionally important for internalizing apoE in human brain. ApoE is associated with β-amyloid in senile plaques and plays a role in the transport of the β-amyloid. The presence of VLDLR in myelin may be one explanation as to why β-amyloid does not accumulate in the white matter which is rich in VLDLR. Recently, evidences on VLDLR obtained mainly using knock-out or transfected mice suggest this receptor to be neither specific for VLDL nor functionally important in mammals. However, no variant form of VLDLR was found in any tissues of mouse. This variant form of VLDLR should thus be studied in greater detail using human tissues or cells.

Extraction and purification of active IgG from SSPE and MS brain

Immunoglobulin (Ig) G was purified from soluble and membrane fractions of postmortem subacute sclerosing panencephalitis (SSPE) brain, multiple sclerosis (MS) brain plaque-periplaque white matter, and normal human brain (NHB) white matter. After homogenization in 0.32 M sucrose and removal of cell debris and nuclei by low-speed centrifugation, soluble and crude membrane fractions were separated by ultracentrifugation. After removal of sucrose by dialysis, IgG was isolated from the soluble fraction by protein A affinity chromatography. IgG was obtained from the membrane fraction by elution at low pH and purification from the eluate by protein A chromatography. Whereas very little IgG was in NHB white matter, significant levels of IgG were recovered from both SSPE and MS brain. Both immunocytochemical staining of measles virus-infected cells in tissue culture and protein immunoblotting of virus-infected cell lysates showed that the IgG from SSPE brain contained activity specific for measles virus protein. The abundance, purity and functional activity of IgG extracted from SSPE and MS brain indicate that IgG extracted from the brain of humans with an inflammatory disease of unknown etiology can be used to identify its corresponding antigen.

Cerebral blood flow change during postcibal hypotension. Comparison between multiple system atrophy and pure autonomic failure

The Diffusion Status of Metabolites in Human Brain: Measured Using Proton Diffusion MR Spectroscopy. Fan~, Rand . Nishitani Rlrnmul, Fllrnkl~u Mivoshi.Ms~afnmi HIrada.Ohl~nka |lidekl. I.Dgpl. of Radiology,XiJing Hospital.The Fourth Military Medical University giaa (710032),P.R.of China 2.Dept. of Radioiog~,,Medicai SchooI,Tokushima University, Tokushima,lapan Mmterl=l, and Methndl: The water suppressed STEAM sequence were used for localized diffusion measurement in 5 normal human brain in viva The diffusion gradients(2.4.and 6 roT/M) were applied during the TFJ2 period. corresponding the b factors as 72,285,and 642 s/ramS.Other parameters including the TE=68ms,TM=200ms,TK = 1625ms and 256 acquisition.The signal intensity of NAA ,Cho,and Cr were measured and the diffusion coefficient (D) were calculated according the equation D=Ln(SIIS0y(bl-bO) [l~nla:The diffusion coefficient of NAA,Cho and Cr were list in follow table table: The Diffusion Coefficient of NAA,Cho and Cr in the normal human brain white matter.(unit:10 -3 mm2/sec)

A stereological method for estimating the total length and size of myelin fibers in human brain white matter

A practically unbiased stereological method to obtain estimates of the volume and total length of nerve fibers in brain white matter is described. The sampling scheme is designed so that the majority of brain white matter is left intact, thus providing the possibility for resampling and further analysis. Uniform sampling of one complete hemispherical white matter is performed. The volume fraction of nerve fibers in white matter is estimated by point counting. The total length of nerve fibers was estimated from the product of the volume of white matter, obtained with the Cavalieri principle, and the fiber length density, obtained from the isotropic, uniform random sections which were ensured by the isector. The size of nerve fibers was derived by measuring the profile diameter perpendicular to its longest axis. The influence of the postmortem fixation delay on nerve fiber parameters was investigated in one dog and one pig. The criteria for identification of nerve fiber profiles at light microscopy were evaluated using electron microscopy.

Strategies to identify sequences or antigens unique to multiple sclerosis.

Chronic inflammatory and infectious diseases of the central nervous system (CNS) are characterized by increased IgG and oligoclonal bands (OGBs) in the brain and cerebrospinal fluid (CSF). The OGBs in CNS infectious diseases of known cause have been shown to be directed against the pathogenic agent. In multiple sclerosis (MS), the antigenic specificity of the OGBs is unknown, but could be directed against an infectious agent, an autoantigen, or both. In a molecular approach to identify antigens specific for MS, we constructed directional cDNA expression libraries with mRNA extracted from chronic and acute MS plaques and periplaque white matter. The libraries were: (1) screened to identify clones whose expression products react with MS CSF, but not with CSF from other infectious and inflammatory diseases of the CNS; (2) subtracted by hybridization to mRNA from normal human brain white matter and differentially screened to detect unique MS transcripts; and (3) used as template in polymerase chain reactions to amplify, clone, and sequence IgG heavy and light chain variable regions (VH and VL, respectively) expressed in MS plaques. Analysis of the VH and VL IgG repertoire in MS brain may identify disease-relevant IgG sequences that can be assembled into functional antibodies using recombinant phage technology. Such recombinant antibodies will be useful to probe brain tissue to identify antigens unique to MS.

Amyloid β Protein and Its 3-kDa Fragment Are Present in the Axoplasm Fraction of the White Matter in Human Brain

Production of the soluble amyloid β-protein (Aβ) precedes abnormal accumulation of Aβ amyloid in the brains of subjects with Alzheimer's disease. To determine the cellular source and generating mechanisms of soluble Aβ in the human brain, we separated an axoplasm fraction from the cerebral white matter and analyzed it. The axoplasm fraction contained secretory isoforms of β-amyloid precursor protein (APP) and 11.5 kDa Aβ-bearing carboxyl-terminal fragments (CTFs) of APP. Furthermore, soluble 4 kDa Aβ and 3 kDa fragments of Aβ (p3) were obtained from the axoplasm fraction. These results suggest that amyloidogenic 4 kDa Aβ is intracellularly produced in cerebral neurons and carried through the axons in human brain.

Identification of a mitogen-activated protein kinase site in human myelin basic protein in situ

Ultrastructural localization of a specific phosphorylated isomer of myelin basic protein (MBP) has been achieved with a monoclonal antibody specific for human MBP sequence, 89–105, in which Thr 98 was phosphorylated. Cryosections of human brain white matter revealed that gold particles were found localized almost exclusively to the major dense line demonstrating that threonine 98 in the sequence Thr-Pro-Arg-Thr-Pro-Pro-Pro, a mitogen-activated protein kinase-specific site, was phosphorylated in vivo. In two cases of multiple sclerosis, the density of gold particles in myelin was reduced by about 30%, in one case by 42%, and by 80% in a fourth case. However, gold labelling was seen in areas of demyelination suggesting that the phosphorylated threonyl peptide was protected from degradation.

Relative Concentrations of Proton MR Visible Neurochemicals in Gray and White Matter in Human Brain

The relative distributions of N-acetylaspartate (NAA) + N-acetylaspartylglutamate (NAAG), creatine + phosphocreatine (Cr/PCr), and choline (Cho) in the gray and white matter of human brain were determined by utilizing proton magnetic resonance spectroscopic imaging (SI). The SI data was processed using an automated spectroscopic image processing algorithm, and image segmentation was performed using a supervised technique. Linear regression analysis indicated that the NAA + NAAG (2.01 ppm) and Cr/PCr (3.02 ppm) peaks are greater in gray matter compared with white matter. The large intersubject variation observed in the Cho (3.20 ppm) resonance prevented the assessment of its regional distribution with confidence.

Secretory form of β-amyloid precursor protein is much abundantly contained in the cerebral white matter in human brain

To elucidate the metabolic process of β-amyloid precursor proteins (APP) in cerebral gray and white matter of the human brain, we compared the content and characteristics of secretory APP between these two parts. The white matter contained much more secretory APP than the gray matter in both a control and a Down's syndrome brain, but no difference in the characteristics of the APP isoforms was detected. Our results suggest that, in the human brain, a considerable amount of APP is carried by axonal transport, during which some of the APP isoforms are processed to their secretory forms.

Molecular species composition of glycerophospholipids from white matter of human brain.

The molecular species composition of the major glycerophospholipids from white matter of human brain were determined by high-performance liquid chromatography of the 3,5-dinitrobenzoyl derivatives of the corresponding diradylglycerols. In phosphatidylcholine (PC) and phosphatidylserine (PS), molecular species containing only saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) comprised 85.7 and 82.4% of the respective totals, with 18:0/18:1 predominant in PS and 16:0/18:1 in PC. These molecular species were also abundant in phosphatidylethanolamine (PE), but in this phospholipid species containing polyunsaturated fatty acids (PUFA), largely 18:0/22:6n-3 and 18:0/20:4n-6, accounted for over half the total; 18:1/18:1 was also abundant in PE. In contrast, 1-O-alk-1'-enyl-2-acyl sn-glycero-3-phosphoethanolamine (GPE) had much more SFA- and MUFA-containing species, predominantly 16:0a/18:1, 18:0a/18:1 and 18:1a/18:1, with low amounts of species containing 20:4n-6 and 22:6n-3. In alkenylacyl GPE, 22:4n-6 was the major PUFA and 16:0a/22:4n-6 and 18:1a/22:4n-6 the main PUFA-containing species. There was six times more 22:6n-3, twice as much 20:4n-6 and half the amount of 22:4n-6 in PE as compared to alkenylacyl GPE.

Quantification of D-aspartate in normal and Alzheimer brains.

Using a new procedure to hydrolyze proteins without provoking racemization of the amino acids and using enzymatic methods to determine D- and L-aspartate (Asp), we have quantified the content of protein-bound D-aspartate (both D-aspartic acid and D-asparagine) of human brain white and gray matter proteins from normal and Alzheimer subjects. The D-enantiomer is present in brain proteins at mean concentrations between 0.48 and 0.90 mumol/g of wet tissue, corresponding to concentrations 34-82 times lower than that of L-aspartate. The highest levels of D-aspartate were found in Alzheimer gray matter (0.60-0.90, mean 0.69 mumol/g of wet tissue). When expressed as the percentage of total (i.e. D- plus L-) aspartate, %D = [D/(D + L)] x 100, the Alzheimer brains show a significantly higher content of D-aspartate in both gray matter (2.08%) and white matter (1.80%) than in the corresponding tissues of normal brains (1.65% in gray, 1.58% in white).

Experimental allergic encephalomyelitis in cynomolgus monkeys. Quantitation of T cell responses in peripheral blood.

Chronic relapsing-remitting experimental allergic encephalomyelitis (EAE) was induced in cynomolgus monkeys by a single immunization with a homogenate of human brain white matter (BH) in adjuvant. Proliferative T lymphocyte responses to BH, to myelin basic protein (MBP), but not to proteolipid protein, were detected in peripheral blood mononuclear cells (PBMC) of all animals and persisted until their death or, in surviving animals, for greater than 10 mo postimmunization. Responses of higher magnitude tended to be associated with fatal, compared with nonfatal, episodes of clinical EAE. The frequency of MBP-reactive T cells in PBMC of animals with acute EAE was quantitated with a soft agar colony system; the ratio of T cells that proliferated specifically to MBP was estimated at between 5 and 20 per 10(6) PBMC. A similar frequency of peptide-specific T cells was estimated from PBMC of monkeys immunized with a synthetic 14-mer peptide corresponding to a region near the carboxy terminus of MBP. Thus, autoantigen-reactive T cells can be detected in the circulation throughout the course of chronic EAE, are predictive of disease severity, and occur at a frequency similar to that estimated to be present in humans with multiple sclerosis.

Plasmalopsychosine, a novel plasmal (fatty aldehyde) conjugate of psychosine with cyclic acetal linkage. Isolation and characterization from human brain white matter.

Through a systematic examination of basic (cationic) lipids separated on Folch's lower phase from extracts of human brain by cation exchange chromatography on carboxymethyl Sephadex in a chloroform/methanol mixture, followed by successive chromatographies on Florisil and Iatrobeads columns, five compounds of basic lipids were separated. Two major unknown compounds A and B and a minor unknown compound C were separated, in addition to minor compounds sphingosine and N,N-dimethylsphingosine. This paper describes the isolation and chemical characterization of major unknown compounds A and B, which were found only in the white matter but not in the gray matter of the human brain. Unmodified psychosine (galactosylsphingosine) was essentially undetectable under the experimental conditions. Unknown compounds A and B were identified as novel plasmal (fatty aldehyde) conjugates of psychosine with cyclic acetal linkage at the galactosyl residue of psychosine. Fatty aldehydes were identified as mainly palmital (16:0) and stearal (18:0). Sphingosine was identified as d18:1 sphingosine. Faster migrating compound A had 3,4-cyclic acetal linkage, and slower migrating compound B had 4,6-cyclic acetal linkage (where m is 14 or 16 and n is 12) as shown below. [formula: see text] Preliminary studies showed that compounds A, B, and C had a weak inhibitory effect on protein kinase C (PKC) and had no cytotoxic effect. In contrast, psychosine displayed a strong cytotoxicity and inhibitory effect on PKC. Therefore, the process controlling the addition or deletion of plasmal cyclic linkage to psychosine could be a crucial step in regulation of PKC, src, or other kinases susceptible to psychosine.