Abstract
Epilepsy, deafness, onychodystrophy, osteodystrophy and intellectual disability are associated with a spectrum of mutations of human TBC1D24. The mechanisms underlying TBC1D24-associated ...disorders and the functions of TBC1D24 are not well understood. Using CRISPR-Cas9 genome editing, we engineered a mouse with a premature translation stop codon equivalent to human S324Tfs*3, a recessive mutation of TBC1D24 associated with early infantile epileptic encephalopathy (EIEE). Homozygous S324Tfs*3 mice have normal auditory and vestibular functions but show an abrupt onset of spontaneous seizures at postnatal day 15 recapitulating human EIEE. The S324Tfs*3 variant is located in an alternatively spliced micro-exon encoding six perfectly conserved amino acids incorporated postnatally into TBC1D24 protein due to a micro-exon utilization switch. During embryonic and early postnatal development, S324Tfs*3 homozygotes produce predominantly the shorter wild-type TBC1D24 protein isoform that omits the micro-exon. S324Tfs*3 homozygotes show an abrupt onset of seizures at P15 that correlates with a developmental switch to utilization of the micro-exon. A mouse deficient for alternative splice factor SRRM3 impairs incorporation of the Tbc1d24 micro-exon. Wild-type Tbc1d24 mRNA is abundantly expressed in the hippocampus using RNAscope in situ hybridization. Immunogold electron microscopy using a TBC1D24-specific antibody revealed that TBC1D24 is associated with clathrin-coated vesicles and synapses of hippocampal neurons, suggesting a crucial role of TBC1D24 in vesicle trafficking important for neuronal signal transmission. This is the first characterization of a mouse model of human TBC1D24-associated EIEE that can now be used to screen for antiepileptogenic drugs ameliorating TBCID24 seizure disorders.
Indices of diffusion anisotropy calculated from diffusion coefficients acquired in two or three perpendicular directions are rotationally variant. In living monkey brain, these indices severely ...underestimate the degree of diffusion anisotropy. New indices calculated from the entire diffusion tensor are rotationally invariant (RI). They show that anisotropy is highly variable in different white matter regions depending on the degree of coherence of fiber tract directions. In structures with a regular, parallel fiber arrangement, water diffusivity in the direction parallel to the fibers (D| ≈ 1400–1800 × 10−6 mm2/s) is almost 10 times higher than the average diffusivity in directions perpendicular to them ((D + D⊥′)/2 ≈ 150–300 × 10−6 mm2/s), and is almost three times higher than previously reported. In structures where the fiber pattern is less coherent (e.g., where fiber bundles merge), diffusion anisotropy is significantly reduced. However, RI anisotropy indices are still susceptible to noise contamination. Monte Carlo simulations show that these indices are statistically biased, particularly those requiring sorting of the eigenvalues of the diffusion tensor based on their magnitude. A new intervoxel anisotropy index is proposed that locally averages inner products between diffusion tensors in neighboring voxels. This “lattice” RI index has an acceptably low error variance and is less susceptible to bias than any other RI anisotropy index proposed to date.
Analytical expressions of the diffusion tensor of water, D, and of scalar invariants derived from it, are given in terms of the intensities of seven diffusion‐weighted images (DWIs). These formulas ...simplify the post‐processing steps required in diffusion tensor imaging, including estimating D in each voxel (from the set of b‐matrices and their corresponding DWIs), and then computing its eigenvalues, eigenvectors, and scalar invariants. In a study conducted using artifact‐free DWIs with high diffusion weighting (bmax ˜ 900 s/mm2), maps of Trace(D) and the Relative and Lattice Anisotropy indices calculated analytically and by multivariate linear regression showed excellent agreement in brain parenchyma of a healthy living cat. However, the quality of the analytically computed maps degraded markedly as diffusion weighting was reduced. Although diffusion tensor MRI with seven DWIs may be useful for clinical applications where rapid scanning and data processing are required, it does not provide estimates of the uncertainty of the measured imaging parameters, rendering it susceptible to noise and systematic artifacts. Therefore, care should be taken when using this technique in radiological applications.
Mild traumatic brain injury (mTBI) is highly prevalent but lacks both research tools with adequate sensitivity to detect cellular alterations that accompany mild injury and pre-clinical models that ...are able to robustly mimic hallmark features of human TBI. To address these related challenges, high-resolution diffusion tensor MRI (DTI) analysis was performed in a model of mild TBI in the ferret - a species that, unlike rodents, share with humans a gyrencephalic cortex and high white matter (WM) volume. A set of DTI image analysis tools were optimized and implemented to explore key features of DTI alterations in
adult male ferret brains (
= 26), evaluated 1 day to 16 weeks after mild controlled cortical impact (CCI). Using template-based ROI analysis, lesion overlay mapping and DTI-driven tensor-based morphometry (D-TBM) significant differences in DTI and morphometric values were found and their dependence on time after injury evaluated. These observations were also qualitatively compared with immunohistochemistry staining of neurons, astrocytes, and microglia in the same tissue. Focal DTI abnormalities including reduced cortical diffusivity were apparent in 12/13 injured brains with greatest lesion extent found acutely following CCI by ROI overlay maps and reduced WM FA in the chronic period was observed near to the CCI site (ANOVA for FA in focal WM: time after CCI
= 0.046, brain hemisphere
= 0.0012) often in regions without other prominent MRI abnormalities. Global abnormalities were also detected, especially for WM regions, which demonstrated reduced diffusivity (ANOVA for Trace: time after CCI
= 0.007) and atrophy that appeared to become more extensive and bilateral with longer time after injury (ANOVA for D-TBM Log of the Jacobian values: time after CCI
= 0.007). The findings of this study extend earlier work in rodent models especially by evaluation of focal WM abnormalities that are not influenced by partial volume effects in the ferret. There is also substantial overlap between DTI and morphometric findings in this model and those from human studies of mTBI implying that the combination of DTI tools with a human-similar model system can provide an advantageous and informative approach for mTBI research.
Pre-clinical models of traumatic brain injury (TBI) have been the primary experimental tool for understanding the potential mechanisms and cellular alterations that follow brain injury, but the human ...relevance and translational value of these models are often called into question. Efforts to better recapitulate injury biomechanics and the use of non-rodent species with neuroanatomical similarities to humans may address these concerns and promise to advance experimental studies toward clinical impact. In addition to improving translational aspects of animal models, it is also advantageous to establish pre-clinical outcomes that can be directly compared with the same outcomes in humans. Non-invasive imaging and particularly MRI is promising for this purpose given that MRI is a primary tool for clinical diagnosis and at the same time increasingly available at the pre-clinical level. The objective of this study was to identify which commonly used radiologic markers of TBI outcomes can be found also in a translationally relevant pre-clinical model of TBI. The ferret was selected as a human relevant species for this study with folded cortical geometry and relatively high white matter content and the closed head injury model of engineered rotation and acceleration (CHIMERA) TBI model was selected for biomechanical similarities to human injury. A comprehensive battery of MRI protocols based on common data elements (CDEs) for human TBI was collected longitudinally for the identification of MRI markers and voxelwise analysis of T2, contrast enhancement and diffusion tensor MRI values. The most prominent MRI findings were consistent with focal hemorrhage and edema in the brain stem region following high severity injury as well as vascular and meningeal injury evident by contrast enhancement. While conventional MRI outcomes were not highly conspicuous in less severe cases, quantitative voxelwise analysis indicated diffusivity and anisotropy alterations in the acute and chronic periods after TBI. The main conclusions of this study support the translational relevance of closed head TBI models in intermediate species and identify brain stem and meningeal vulnerability. Additionally, the MRI findings highlight a subset of CDEs with promise to bridge pre-clinical studies with human TBI outcomes.
In this work we report findings from an in vivo diffusion tensor imaging (DTI) study of the human optic chiasm at sub-millimeter voxel resolution. Data were collected at 3 T using a ...diffusion-weighted radial-FSE sequence, which provides images free from typical magnetic susceptibility artifacts. The general DTI features observed in the optic chiasm region were consistent across subjects. They included a central area with high anisotropy and highest diffusivity in a predominately right/left direction corresponding to the decussation of nasal hemiretinae fibers, surrounded by a band of low anisotropy reflecting heterogeneous orientation of fibers within the voxel, and a lateral area with high anisotropy and highest diffusivity in a predominately anterior/posterior direction corresponding to temporal hemiretinae fibers that do not cross. Animal studies indicate that there is a significant dorsal–ventral reorganization of the retinotopic distribution of fibers along the optic pathways. We found that diffusion ellipsoids in the central portion of the optic chiasm show considerable planar anisotropy in the coronal plane indicating fiber crossings in the superior/inferior direction, rather than strictly right/left. This architectural feature of the chiasm suggests that dorso–ventral reorganization of fibers in the optic pathways also occurs in humans. We have shown that by collecting sub-millimeter resolution data, DTI can be used to investigate fine details of small and complex white matter structures, in vivo, with a clinical scanner. High spatial resolution, however, is necessary in the slice direction as well as in-plane to reduce the CSF contribution to the signal and to increase fiber coherence within voxels.
Magnetic resonance diffusion imaging is potentially an important tool for the noninvasive characterization of normal and pathological tissue. The technique, however, is prone to a number of artifacts ...that can severely affect its ability to provide clinically useful information. In this study, the problem of eddy current‐induced geometric distortions that occur in diffusion images acquired with echo planar sequences was addressed. These geometric distortions produce artifacts in computed maps of diffusion parameters and are caused by misalignments in the individual diffusion‐weighted images that comprise the diffusion data set. A new approach is presented to characterize and calibrate the eddy current effects, enabling the eddy current distortions to be corrected in sets of Interleaved (or snapshot) echo planar diffusion images. Correction is achieved by acquiring one‐dimensional field maps in the read and phase encode direction for each slice and each diffusion step. The method is then demonstrated through the correction of distortions in diffusion images of the human brain. It is shown that by using the eddy current correction scheme outlined, the eddy current‐induced artifacts in the diffusion‐weighted images are almost completely eliminated. In addition, there is a significant improvement in the quality of the resulting diffusion tensor maps.
Diffusion tensor MRI tractography aims to reconstruct noninvasively the 3D trajectories of white matter fasciculi within the brain, providing neuroscientists and clinicians with a potentially useful ...tool for mapping brain architecture. While this technique is widely used to visualize white matter pathways, the associated uncertainty in fiber orientation and artifacts have, to date, not been visualized in conjunction with the trajectory data. In this work, the bootstrap method was used to determine the distributions of diffusion indices such as trace and anisotropy, together with the uncertainty in fiber orientation. A novel visualization scheme was developed to encode this information at each point along reconstructed trajectories. By integrating these schemes into a graphical user interface, a new tool which we call PASTA (Pointwise Assessment of Streamline Tractography Attributes) was created to facilitate identification of artifacts in tractography that would otherwise go undetected. Magn Reson Med 53:1462–1467, 2005. Published 2005 Wiley‐Liss, Inc.
Little is known about genetic regulation of the development of white matter. This knowledge is critical in understanding the pathophysiology of neurodevelopmental syndromes associated with altered ...cognition as well as in elucidating the genetics of normal human cognition. The hemideletion of almost equal to25 genes on chromosome 7q11.23 that causes Williams syndrome (WS) includes genes that regulate cytoskeletal dynamics in neurons, especially LIMK1 and CYLN2, and therefore offers the opportunity to investigate the role of these genes in the formation of white matter tracts. We used diffusion tensor imaging to demonstrate alteration in white matter fiber directionality, deviation in posterior fiber tract course, and reduced lateralization of fiber coherence in WS. These abnormalities are consistent with an alteration of the late stages of neuronal migration, define alterations of white matter structures underlying dissociable behavioral phenotypes in WS, and provide human in vivo information about genetic control of white matter tract formation.