Hao Huang

Stereotaxic White Matter Atlas Based on Diffusion Tensor Imaging in an ICBM Template

Brain registration to a stereotaxic atlas is an effective way to report anatomic locations of interest and to perform anatomic quantification. However, existing stereotaxic atlases lack comprehensive coordinate information about white matter structures. In this paper, white matter-specific atlases in stereotaxic coordinates are introduced. As a reference template, the widely used ICBM-152 was used. The atlas contains fiber orientation maps and hand-segmented white matter parcellation maps based on diffusion tensor imaging (DTI). Registration accuracy by linear and non-linear transformation was measured, and automated template-based white matter parcellation was tested. The results showed a high correlation between the manual ROI-based and the automated approaches for normal adult populations. The atlases are freely available and believed to be a useful resource as a target template and for automated parcellation methods.

White and gray matter development in human fetal, newborn and pediatric brains

Brain anatomy is characterized by dramatic growth from the end of the second trimester through the neonatal stage. The characterization of normal axonal growth of the white matter tracts has not been well-documented to date and could provide important clues to understanding the extensive inhomogeneity of white matter injuries in cerebral palsy (CP) patients. However, anatomical studies of human brain development during this period are surprisingly scarce and histology-based atlases have become available only recently. Diffusion tensor magnetic resonance imaging (DTMRI) can reveal detailed anatomy of white matter. We acquired diffusion tensor images (DTI) of postmortem fetal brain samples and in vivo neonates and children. Neural structures were annotated in two-dimensional (2D) slices, segmented, measured, and reconstructed three-dimensionally (3D). The growth status of various white matter tracts was evaluated on cross-sections at 19-20 gestational weeks, and compared with 0-month-old neonates and 5- to 6-year-old children. Limbic, commissural, association, and projection white matter tracts and gray matter structures were illustrated in 3D and quantitatively characterized to assess their dynamic changes. The overall pattern of the time courses for the development of different white matter is that limbic fibers develop first and association fibers last and commissural and projection fibers are forming from anterior to posterior part of the brain. The resultant DTMRI-based 3D human brain data will be a valuable resource for human brain developmental study and will provide reference standards for diagnostic radiology of premature newborns.

Human Brain White Matter Atlas: Identification and Assignment of Common Anatomical Structures in Superficial White Matter

Structural delineation and assignment are the fundamental steps in understanding the anatomy of the human brain. The white matter has been structurally defined in the past only at its core regions (deep white matter). However, the most peripheral white matter areas, which are interleaved between the cortex and the deep white matter, have lacked clear anatomical definitions and parcellations. We used axonal fiber alignment information from diffusion tensor imaging (DTI) to delineate the peripheral white matter, and investigated its relationship with the cortex and the deep white matter. Using DTI data from 81 healthy subjects, we identified nine common, blade-like anatomical regions, which were further parcellated into 21 subregions based on the cortical anatomy. Four short association fiber tracts connecting adjacent gyri (U-fibers) were also identified reproducibly among the healthy population. We anticipate that this atlas will be useful resource for atlas-based white matter anatomical studies.

DTI tractography based parcellation of white matter: application to the mid-sagittal morphology of corpus callosum.

Morphology of the corpus callosum (CC) at the mid-sagittal level has been a target of extensive studies. However, the lack of internal structures and its polymorphism make it a challenging task to quantitatively analyze shape differences among subjects. In this paper, diffusion tensor Imaging (DTI) and tract tracing technique were applied to incorporate cortical connectivity information to the morphological study. The CC was parcellated into six major subdivisions based on trajectories to different cortical areas. This subdivision was performed for eight normal subjects and one stroke patient. The parcellated CCs of the normal subjects were normalized for morphological analysis. When comparing the stroke patient to the normal population, we detected significant atrophy in the motor and sensory areas of the patient CC, in line with the clinical deficits. This approach provides a new tool to investigate callosal morphology and functional relationships.

Maternal infection leads to abnormal gene regulation and brain atrophy in mouse offspring: implications for genesis of neurodevelopmental disorders.

Prenatal viral infection has been associated with development of schizophrenia and autism. Our laboratory has previously shown that viral infection causes deleterious effects on brain structure and function in mouse offspring following late first trimester (E9) administration of influenza virus. We hypothesized that late second trimester infection (E18) in mice may lead to a different pattern of brain gene expression and structural defects in the developing offspring. C57BL6J mice were infected on E18 with a sublethal dose of human influenza virus or sham-infected using vehicle solution. Male offsping of the infected mice were collected at P0, P14, P35 and P56, their brains removed and prefrontal cortex, hippocampus and cerebellum dissected and flash frozen. Microarray, qRT-PCR, DTI and MRI scanning, western blotting and neurochemical analysis were performed to detect differences in gene expression and brain atrophy. Expression of several genes associated with schizophrenia or autism including Sema3a, Trfr2 and Vldlr were found to be altered as were protein levels of Foxp2. E18 infection of C57BL6J mice with a sublethal dose of human influenza virus led to significant gene alterations in frontal, hippocampal and cerebellar cortices of developing mouse progeny. Brain imaging revealed significant atrophy in several brain areas and white matter thinning in corpus callosum. Finally, neurochemical analysis revealed significantly altered levels of serotonin (P14, P35), 5-Hydroxyindoleacetic acid (P14) and taurine (P35). We propose that maternal infection in mouse provides an heuristic animal model for studying the environmental contributions to genesis of schizophrenia and autism, two important examples of neurodevelopmental disorders.

Analysis of noise effects on DTI-based tractography using the brute-force and multi-ROI approach

Diffusion tensor tractography based on line propagation is a promising and widely used technique, but it is known to be sensitive to noise and the size and location of the seed regions of interest (ROIs). The effects of these parameters on the tractography results were analyzed quantitatively using high‐resolution diffusion tensor imaging (DTI) with a high signal‐to‐noise ratio (SNR) on a fixed mouse brain. The anterior commissure (AC), as judged from a T2‐weighted image, was used as an anatomical reference within which the tracts could be located. Monte Carlo simulation was performed by adding Gaussian noise to the time domain data and repeating the tractography. Deviations of the tracking results were measured as a function of SNR. Such noise effects were evaluated for a simple one‐ROI approach and a combined two‐ROI and brute‐force (BF) approach. The influence of ROI size and location for the two‐ROI + BF approach was also analyzed. The results confirmed the hypothesis that one can increase the validity of DTI‐based tractography by adopting the BF and multi‐ROI approach, with respect to the simple one‐ROI approach. Magn Reson Med 52:559–565, 2004.

Three-dimensional anatomical characterization of the developing mouse brain by diffusion tensor microimaging.

Investigation of three-dimensional (3D) morphometry of developing brains has been hindered by a lack of imaging modalities that can monitor the 3D evolution of various anatomical structures without sectioning and staining processes. In this study, we combined magnetic resonance microimaging and diffusion tensor imaging techniques to accomplish such visualization. The application of this approach to developing mouse embryos revealed that it could clearly delineate early critical structures such as neuroepithelium, cortical plate, and various axonal structures, and follow their developmental evolution. The technique was applied to the study of the Netrin-1 mutant, allowing verification of its anatomical phenotype.

Development of axonal pathways in the human fetal fronto-limbic brain: histochemical characterization and diffusion tensor imaging

The development of cortical axonal pathways in the human brain begins during the transition between the embryonic and fetal period, happens in a series of sequential events, and leads to the establishment of major long trajectories by the neonatal period. We have correlated histochemical markers (acetylcholinesterase (AChE) histochemistry, antibody against synaptic protein SNAP‐25 (SNAP‐25‐immunoreactivity) and neurofilament 200) with the diffusion tensor imaging (DTI) database in order to make a reconstruction of the origin, growth pattern and termination of the pathways in the period between 8 and 34 postconceptual weeks (PCW). Histological sections revealed that the initial outgrowth and formation of joined trajectories of subcortico‐frontal pathways (external capsule, cerebral stalk–internal capsule) and limbic bundles (fornix, stria terminalis, amygdaloid radiation) occur by 10 PCW. As early as 11 PCW, major afferent fibers invade the corticostriatal junction. At 13–14 PCW, axonal pathways from the thalamus and basal forebrain approach the deep moiety of the cortical plate, causing the first lamination. The period between 15 and 18 PCW is dominated by elaboration of the periventricular crossroads, sagittal strata and spread of fibers in the subplate and marginal zone. Tracing of fibers in the subplate with DTI is unsuccessful due to the isotropy of this zone. Penetration of the cortical plate occurs after 24–26 PCW. In conclusion, frontal axonal pathways form the periventricular crossroads, sagittal strata and ‘waiting’ compartments during the path‐finding and penetration of the cortical plate. Histochemistry is advantageous in the demonstration of a growth pattern, whereas DTI is unique for demonstrating axonal trajectories. The complexity of fibers is the biological substrate of selective vulnerability of the fetal white matter.

Abnormal expression of myelination genes and alterations in white matter fractional anisotropy following prenatal viral influenza infection at E16 in mice

Prenatal viral infection has been associated with the development of schizophrenia and autism. Our laboratory has previously shown that viral infection causes deleterious effects on brain structure and function in mouse offspring following late first trimester (E9) and late second trimester (E18) administration of influenza virus. We hypothesized that middle second trimester infection (E16) in mice may lead to a different pattern of brain gene expression and structural defects in the developing offspring. C57BL6 mice were infected on E16 with a sublethal dose of human influenza virus or sham-infected using vehicle solution. Male offspring of the infected mice were collected at P0, P14, P35, and P56, their brains removed and cerebella dissected and flash frozen. Microarray, DTI and MRI scanning, as well as qRT-PCR and SDS-PAGE and western blotting analyses were performed to detect differences in gene expression and brain atrophy. Expression of several genes associated with myelination, including Mbp, Mag, and Plp1 were found to be altered, as were protein levels of Mbp, Mag, and DM20. Brain imaging revealed significant atrophy in cerebellum at P14, reduced fractional anisotropy in white matter of the right internal capsule at P0, and increased fractional anisotropy in white matter in corpus callosum at P14 and right middle cerebellar peduncle at P56. We propose that maternal infection in mouse impacts myelination genes.

Evidence of slow maturation of the superior longitudinal fasciculus in early childhood by diffusion tensor imaging.

While the majority of axonal organization is established by birth in mammalian brains, axonal wiring and pruning processes, as well as myelination, are known to extend to the postnatal periods, where environmental stimuli often play a major role. Normal axonal and myelin development of individual white matter tracts of human in this period is poorly understood and may have a major role in cognitive development of human. In this study, we applied diffusion tensor imaging and normalization-based population analyses to 44 preteen children and 30 adult images. We observed highly significant changes of fiber orientations at regions that correspond to the superior longitudinal fasciculus during the first 5 years. The result is attributed to slow axonal and/or myelin maturation of this tract, which is believed to be involved in language functions.