ABSTRACT
Diffusion Tensor Imaging (DTI) and Manganese Enhanced MRI (MEMRI) are noninvasive tools to characterize neural fiber microstructure and axonal transport. A combination of both may provide novel insights into the progress of neurodegeneration. To investigate the relationship of DTI and MEMRI in white matter of tauopathy, twelve optic nerves of 11-month-old p301L tau mice were imaged and finished with postmortem immunohistochemistry. MEMRI was used to quantify Mn2+ accumulation rates in the optic nerve (ON, termed ONAR) and the Superior Colliculus (SC, termed SCAR), the primary terminal site of ON in mice. We found that both ONAR and SCAR revealed a significant linear correlation with mean diffusion (mD) and radial diffusion (rD) but not with other DTI quantities. Immunohistochemistry findings showed that ONAR, mD, and rD are significantly correlated with the myelin content (Myelin Basic Protein, p < 0.05) but not with the axonal density (SMI-31), tubulin density, or tau aggregates (AT8 staining). In summary, slower axonal transport appeared to have less myelinated axons and thinner remaining axons, associated with reduced rD and mD of in vivo DTI. A combination of in vivo MEMRI and DTI can provide critical information to delineate the progress of white matter deficits in neurodegenerative diseases.
Subject(s)
Tauopathies , White Matter , Animals , Axonal Transport , Diffusion Tensor Imaging , Disease Models, Animal , MiceABSTRACT
White matter abnormalities, revealed by Diffusion Tensor Imaging (DTI), are observed in patients with Alzheimer's Disease (AD), representing neural network deficits that underlie gradual cognitive decline in patients. However, how DTI changes related to the development of Amyloid beta (Aß) and tau pathology, two key hallmarks of AD, remain elusive. We hypothesized that tauopathy induced by Aß could initiate an axonal degeneration, leading to DTI-detectable white matter abnormalities. We utilized the visual system of the transgenic p301L tau mice as a model system. Aß was injected in Lateral Geniculate Nucleus (LGN), where the Retinal Ganglion Cell (RGC) axons terminate. Longitudinal DTI was conducted to detect changes in the optic tract (OT) and optic nerve (ON), containing the distal and proximal segments of RGC axons, respectively. Our results showed DTI changes in OT (significant 13.2% reduction in axial diffusion, AxD vs. vehicle controls) followed by significant alterations in ON AxD and fractional anisotropy, FA. Histology data revealed loss of synapses, RGC axons and cell bodies resulting from the Aß injection. We further tested whether microtubule-stabilizing compound Epothilone D (EpoD) could ameliorate the damage. EpoD co-treatment with Aß was sufficient to prevent Aß-induced axon and cell loss. Using an acute injection paradigm, our data suggest that EpoD may mediate its protective effect by blocking localized, acute Aß-induced tau phosphorylation. This study demonstrates white matter disruption resulting from localized Aß, the importance of tau pathology induction to changes in white matter connectivity, and the use of EpoD as a potential therapeutic avenue to prevent the axon loss in AD.
Subject(s)
Amyloid beta-Peptides/pharmacology , Epothilones/pharmacology , Geniculate Bodies/drug effects , Nerve Degeneration , Peptide Fragments/pharmacology , Retinal Ganglion Cells/drug effects , Tauopathies , Tubulin Modulators/pharmacology , White Matter , Amyloid beta-Peptides/administration & dosage , Animals , Diffusion Tensor Imaging , Disease Models, Animal , Epothilones/administration & dosage , Mice , Nerve Degeneration/chemically induced , Nerve Degeneration/diagnostic imaging , Nerve Degeneration/drug therapy , Nerve Degeneration/prevention & control , Peptide Fragments/administration & dosage , Tauopathies/chemically induced , Tauopathies/diagnostic imaging , Tauopathies/drug therapy , Tauopathies/pathology , Tubulin Modulators/administration & dosage , White Matter/diagnostic imaging , White Matter/drug effectsABSTRACT
BACKGROUND: Clinical imaging modalities including optical coherence tomography (OCT) and diffusion tensor imaging (DTI) are vital in Multiple Sclerosis (MS), but their relationships during the different phases of Retinal ganglion cell (RGC) degeneration are not clear. We hypothesize that initial injury in optic nerve causes axonal degeneration leading to RGC loss in retina, which can be characterized by a combination of DTI and OCT. Our objective was to examine the correlation between noninvasive and histological data to chronicle the degeneration profile of RGCs in the retina and optic nerve in a mouse model of MS. MATERIALS AND METHODS: Experimental Autoimmune Encephalomyelitis (EAE) was induced in 11 C57Bl/6 mice, with 8 mice reserved as controls. OCT and DTI was conducted 2-8 weeks after induction of EAE. The thickness of the retinal ganglion cell complex (GCC) was measured using OCT and compared to DTI indices measured in optic nerves. End-stage histology was used to quantify axon/myelin loss in the optic nerve and retinal thinning/RGC loss in the retina. RESULTS: Significant changes in DTI-derived Axial Diffusivity (AD, -17.2%) and Trace Diffusivity (TR, -18.3%) began after 2 weeks of EAE. Later significant reductions in Fractional Anisotropy (FA) and AD, with increases in Radial Diffusion (RD) were apparent after 4 and 8 weeks. OCT-derived measures of GCC thickness were reduced after 4 weeks, and reached significant reduction after 8 weeks. Among EAE mice, DTI (FA, AD and RD measures) and OCT measures were all significantly correlated after 4 and 8 weeks. Among histology measures, RGC density (-23%), RGC size (-27%), and the number of SMI31+ axons (-54%) were reduced significantly. DTI measures of FA and AD along with GCC thinning were the best independent predictors of axon loss. CONCLUSIONS: DTI and OCT measures are tightly correlated during the chronic phase of axonal degeneration (4-8 weeks) in EAE mice. After 8 weeks of EAE, both OCT and DTI measures are strong predictors of axon loss in the Optic Nerve.
Subject(s)
Diffusion Tensor Imaging , Encephalomyelitis, Autoimmune, Experimental/complications , Multiple Sclerosis/complications , Retinal Degeneration/diagnostic imaging , Retinal Ganglion Cells/pathology , Tomography, Optical Coherence , Animals , Axons/pathology , Disease Models, Animal , Female , Mice, Inbred C57BL , Optic Nerve/diagnostic imaging , Optic Nerve/pathology , Retinal Degeneration/etiologyABSTRACT
PURPOSE: Diffusion tensor imaging (DTI) is commonly used to evaluate white matter integrity in multiple sclerosis (MS), but the relationship between DTI measures and functional changes during disease remains ambiguous. Using a mouse model of MS, we tested the hypothesis that DTI measures would correlate to the visual evoked potential (VEPs) dynamically at different disease stages. METHODS: In vivo DTI, gadolinium-enhanced T1WI (Gd-T1WI) and VEPs were performed in 5 control and 25 mice after 2-12 weeks of experimental autoimmune encephalomyelitis (EAE). DTI indices, including fractional anisotropy (FA), axial and radial diffusivities (AD and RD), and Gd-T1WI enhancement, were measured in the optic nerve and tract (ON and OT), which were compared with measured VEPs. RESULTS: Gd-T1WI showed a 3- to 4-fold enhancement over controls beginning after 2 weeks of EAE. Across the time course, we found progressive reductions in FA and increases in RD with increases in VEP latency and reductions in amplitude. Significant correlations between DTI (FA and RD) and VEP evolved; in control/early asymptomatic EAE mice, both FA and RD were highly correlated with VEP latency (but not amplitude), while in late EAE, both DTI indices were highly correlated with VEP amplitude (but not latency). CONCLUSION: DTI measures FA and RD are associated to VEP latency in early stages of EAE but associated to VEP amplitude in later stages, suggesting that the patterns of DTI related to the functional decline may depend on the stage of disease progression.
Subject(s)
Diffusion Tensor Imaging/methods , Evoked Potentials, Visual , Multiple Sclerosis/diagnostic imaging , Multiple Sclerosis/physiopathology , Optic Nerve/diagnostic imaging , Optic Nerve/physiopathology , Animals , Contrast Media , Disease Models, Animal , Disease Progression , Electrophysiology/methods , Female , Gadolinium DTPA , Image Processing, Computer-Assisted , Mice , Mice, Inbred C57BL , White MatterABSTRACT
PURPOSE: To evaluate the feasibility of using diffusion tensor imaging (DTI) to characterize the temporospatial profile of axonal degeneration and its relation to blood-brain barrier (BBB) permeability. MATERIALS AND METHODS: Longitudinal DTI was performed in Wallerian degeneration slow (WldS) mice following retinal ischemia. In parallel, gadolinium (Gd)-enhanced T1 -weighted imaging (Gd-T1 WI) was performed to evaluate BBB permeability in white matter during axonal degeneration. To confirm the in vivo findings, immunohistochemistry using SMI-31 and myelin basic protein (MBP) was performed to examine the axons and myelin, respectively, and Evans blue was used to evaluate the permeability of the BBB. RESULTS: Reduced axial diffusivity was found in the optic nerve (ON, -15%, P = 0.0063) 1 week and optic tact (OT, -18%, P = 0.0077) 2 weeks after retinal ischemia, which were respectively associated with an 11% (P = 0.0116) and 25% (P = 0.0001) axonal loss. Increased radial diffusivity was found 1-2 weeks after the colocated decrease of axial diffusivity (35% increase, P = 0.0388 in the ON at week 2 and an 80% increase, P = 0.0015 in the OT at week 4). No significant changes were observed using Gd-T1 WI (P = 0.13-0.75), although an approximately 1-fold increase in Evans blue staining intensity was found in the injured ON and OT starting 1 week after retinal ischemia. CONCLUSION: We demonstrated the utility of DTI to characterize anterograde-propagating axonal degeneration through the ON and OT following retinal damage. Evans blue staining revealed serum albumin accumulation at injured sites, although there was no BBB leakage detectable using Gd-T1 WI. LEVEL OF EVIDENCE: 2 J. Magn. Reson. Imaging 2017;45:482-491.
Subject(s)
Axons/pathology , Blood-Brain Barrier/pathology , Diffuse Axonal Injury/pathology , Diffusion Tensor Imaging/methods , Retinal Degeneration/diagnostic imaging , Retinal Degeneration/pathology , Visual Pathways/pathology , Animals , Blood-Brain Barrier/diagnostic imaging , Diffuse Axonal Injury/diagnostic imaging , Feasibility Studies , Female , Mice , Reproducibility of Results , Sensitivity and Specificity , Visual Pathways/diagnostic imaging , Wallerian Degeneration/diagnostic imaging , Wallerian Degeneration/pathologyABSTRACT
BACKGROUND: Synaptic deficits and neuronal loss are the major pathological manifestations of Alzheimer's disease. However, the link between the early synaptic loss and subsequent neurodegeneration is not entirely clear. Cell culture studies have shown that amyloid-ß (Aß) applied to axonal terminals can cause retrograde degeneration leading to the neuronal loss, but this process has not been demonstrated in live animals. OBJECTIVE: To test if Aß applied to retinal ganglion cell axonal terminals can induce axonal damage in the optic nerve and optic tract in mice. METHODS: Aß was injected into the terminal field of the optic tract, in the left lateral geniculate nucleus of wildtype C57BL/6 mice. Following the injection, monthly diffusion tensor imaging was performed. Three months after the injection, mice underwent visual evoked potential recordings, and then sacrificed for immunohistochemical examination. RESULTS: There were no significant changes seen with diffusion tensor imaging in the optic nerve and optic tract 3 months after the Aß injection. The myelin and axons in these regions remained intact according to immunohistochemistry. The only significant changes observed in this study were delayed transduction and reduced amplitude of visual evoked potentials, although both Aß and its reversed form caused similar changes. CONCLUSION: Despite the published in vitro studies, there was no significant axonal damage in the optic nerve and optic tract after injecting Aß onto retinal ganglion cell axonal terminals of wildtype C57BL/6 mice.
Subject(s)
Amyloid beta-Peptides/toxicity , Axons/physiology , Optic Nerve/physiopathology , Optic Tract/physiopathology , Peptide Fragments/toxicity , Retrograde Degeneration/physiopathology , Animals , Axons/pathology , Diffusion Tensor Imaging , Evoked Potentials, Visual , Female , Geniculate Bodies/pathology , Geniculate Bodies/physiopathology , Humans , Immunohistochemistry , Mice, Inbred C57BL , Microelectrodes , Myelin Sheath/pathology , Myelin Sheath/physiology , Optic Nerve/pathology , Optic Tract/pathology , Retrograde Degeneration/pathology , Visual Cortex/pathology , Visual Cortex/physiopathology , Visual Perception/physiologyABSTRACT
BACKGROUND: Diffusion tensor imaging (DTI) suggests the presence of white matter abnormality at the prodromal stage in human Alzheimer's disease (AD). OBJECTIVE: To use a mouse model of AD to determine whether the white matter abnormality detected by in vivo DTI is associated with functional deficits and axon damage. METHODS: Amyloid-ß1-42 (Aß1-42) was injected into the left lateral ventricle in mice. Two months after the injection, in vivo DTI and visual evoked potential (VEP) recordings were performed, followed by immunohistochemistry of phosphorylated neurofilament and myelin basic protein. RESULTS: DTI of Aß1-42-treated mice showed a significant increase of radial diffusivity in white matter including the optic nerves and tracts. The abnormality was associated with decreased amplitude and increased latency of VEP. Immunohistochemistry confirmed a significant loss of axons and myelin integrity. CONCLUSION: White matter damage induced by Aß1-42 in mice can be detected non-invasively by DTI.
Subject(s)
Amyloid beta-Peptides/toxicity , Diffusion Magnetic Resonance Imaging/methods , Leukoencephalopathies/chemically induced , Leukoencephalopathies/diagnosis , Peptide Fragments/toxicity , Animals , Body Weight/drug effects , Disease Models, Animal , Evoked Potentials, Visual/drug effects , Female , Functional Laterality , Leukoencephalopathies/physiopathology , Male , Mice , Mice, Inbred C57BL , Myelin Basic Protein/metabolism , Neurofilament Proteins/metabolism , Optic Nerve/pathologyABSTRACT
PURPOSE: Optic nerve degeneration in diseases such as glaucoma and multiple sclerosis evolves in months to years. The use of Mn(2+)-Enhanced Magnetic Resonance Imaging (MEMRI) in a time-course study may provide new insights into the disease progression. Previously, we demonstrated the feasibility of using a topical administration for Mn(2+) delivery to the visual system. This study is to evaluate the impact of biweekly or monthly repeated Mn(2+) topical administration and the pH levels of the Mn(2+) solutions for MEMRI on the mouse visual pathway. METHODS: Using groups of mice, the MEMRI with an acidic or pH neutralized 1 M MnCl(2) solution was performed. To evaluate the feasibility of repeated MEMRIs, topical-loaded MEMRI was conducted biweekly seven times or monthly three times. The enhancement of MEMRI in the visual system was quantified. After repeated MEMRIs, the corneas were examined by optical coherence tomography. The retinal ganglion cells (RGCs) and optic nerves were examined by histology. RESULTS: All mice exhibited consistent enhancements along the visual system following repeated MEMRIs. The acidic Mn(2+) solution induced a greater MEMRI enhancement as compared with a neutral pH Mn(2+) solution. Significant 20% RGC loss was found after three biweekly Mn(2+) inductions, but no RGC loss was found after three monthly Mn(2+) treatments. The corneal thickness was found increased after seven biweekly topical-loaded MEMRI. CONCLUSIONS: Acidic Mn(2+) solutions enhanced the uptake of Mn(2+) observed on the MEMRI. Increasing the time intervals of repeated Mn(2+) topical administration reduced the adverse effects caused by MEMRI.
Subject(s)
Contrast Media , Image Enhancement/methods , Magnetic Resonance Imaging/methods , Manganese Compounds , Administration, Topical , Animals , Contrast Media/adverse effects , Contrast Media/chemistry , Female , Longitudinal Studies , Manganese Compounds/adverse effects , Manganese Compounds/chemistry , Mice , Mice, Inbred C57BL , Optic Nerve/metabolism , Optic Nerve/pathology , Retinal Ganglion Cells/metabolism , Retinal Ganglion Cells/pathology , Time Factors , Tomography, Optical Coherence , Visual Pathways/metabolism , Visual Pathways/pathologyABSTRACT
PURPOSE: To evaluate topical loading as an alternative to intravitreal injection for Mn(2+)-enhanced magnetic resonance imaging (MEMRI) of the visual system. METHODS: Topical administration of 0.5 to 1.5 M MnCl(2) and intravitreal injections with 0.5 µL 100 mM and 2 µL 1 M MnCl(2) for mouse MEMRI were conducted, followed by immunohistochemistry. In another mouse group, two topical administrations of 1 M Mn(2+) were applied to the same animals 7 days apart, to evaluate the use of MEMRI in a time course study. Dynamic imaging was also conducted to reveal how Mn(2+) travels to the retina. MEMRI with topically loaded MnCl(2) was also conducted in eyes with retinal ischemia, to evaluate whether the enhancements required healthy neurons. RESULTS: After 1 day, topical administration of 1 M and 1.5 M MnCl(2) rendered significant signal enhancement (up to 20%) in the superior colliculus (P < 0.05) that was equivalent to that of the 2-µL 1 M injection. Repeated exposure to Mn(2+) showed reproduced enhancement. Dynamic imaging showed significant enhancement in the iris, retina, and lens boundary, but not in the vitreous space. In retinal ischemic eyes, no enhancement of MEMRI was detected in the optic nerves. The immunohistochemistry of the optic nerve (1.5 mm anterior to the chiasm) and retina showed no injury 1 week after Mn(2+) topical administrations to each mouse. CONCLUSIONS: The results demonstrated the feasibility of using topical administration of Mn(2+) for MEMRI. Topically loaded Mn(2+) did not diffuse into the vitreous space, but was it may have been absorbed into the iris to diffuse or travel via the capillary circulation to reach the retina.
Subject(s)
Chlorides , Diffusion Magnetic Resonance Imaging/methods , Manganese Compounds , Visual Pathways/metabolism , Administration, Topical , Animals , Chlorides/administration & dosage , Feasibility Studies , Female , Geniculate Bodies/metabolism , Image Enhancement/methods , Immunohistochemistry , Intravitreal Injections , Manganese Compounds/administration & dosage , Mice , Mice, Inbred C57BL , Optic Nerve/metabolism , Retina/metabolism , Superior Colliculi/metabolismABSTRACT
OBJECTIVE: Correlation of diffusion tensor imaging (DTI) with histochemical staining for demyelination and axonal damage in multiple sclerosis (MS) ex vivo human cervical spinal cords. BACKGROUND: In MS, demyelination, axonal degeneration, and inflammation contribute to disease pathogenesis to variable degrees. Based upon in vivo animal studies with acute injury and histopathologic correlation, we hypothesized that DTI can differentiate between axonal and myelin pathologies within humans. METHODS: DTI was performed at 4.7 T on 9 MS and 5 normal control fixed cervical spinal cord blocks following autopsy. Sections were then stained for Luxol fast blue (LFB), Bielschowsky silver, and hematoxylin and eosin (H&E). Regions of interest (ROIs) were graded semi-quantitatively as normal myelination, mild (<50%) demyelination, or moderate-severe (>50%) demyelination. Corresponding axonal counts were manually determined on Bielschowsky silver. ROIs were mapped to co-registered DTI parameter slices. DTI parameters evaluated included standard quantitative assessments of apparent diffusion coefficient (ADC), relative anisotropy (RA), axial diffusivity and radial diffusivity. Statistical correlations were made between histochemical gradings and DTI parameters using linear mixed models. RESULTS: Within ROIs in MS subjects, increased radial diffusivity distinguished worsening severities of demyelination. Relative anisotropy was decreased in the setting of moderate-severe demyelination compared to normal areas and areas of mild demyelination. Radial diffusivity, ADC, and RA became increasingly altered within quartiles of worsening axonal counts. Axial diffusivity did not correlate with axonal density (p=0.091). CONCLUSIONS: Increased radial diffusivity can serve as a surrogate for demyelination. However, radial diffusivity was also altered with axon injury, suggesting that this measure is not pathologically specific within chronic human MS tissue. We propose that radial diffusivity can serve as a marker of overall tissue integrity within chronic MS lesions. This study provides pathologic foundation for on-going in vivo DTI studies in MS.
Subject(s)
Image Interpretation, Computer-Assisted/methods , Magnetic Resonance Imaging/methods , Multiple Sclerosis/complications , Multiple Sclerosis/pathology , Nerve Fibers, Myelinated/pathology , Retrograde Degeneration/etiology , Retrograde Degeneration/pathology , Spinal Cord/pathology , Female , Humans , Male , Middle Aged , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
A quantitative magnetization transfer (qMT) technique was employed to quantify the ratio of the sizes of the bound and free water proton pools in ex vivo mouse brains. The goal was to determine the pool size ratio sensitivity to myelin. Fixed brains from both shiverer mice and control littermates were imaged. The pool size ratio in the corpus callosum of shiverer mice was substantially lower than that in the control mice, while there was no distinguishable difference in the pool size ratio in the gray matter. These results correlate with diffusion tensor imaging (DTI) derived radial diffusivity which previously was shown to reflect myelin integrity in this animal model. Histological study reveals the presence of myelin in control mice white matter and the absence of myelin in shiverer mice white matter, supporting the qMT and DTI results. Our findings support the view that qMT may be used for estimating myelin integrity.
Subject(s)
Diffusion Magnetic Resonance Imaging/methods , Magnetics , Myelin Sheath/pathology , Shivering/physiology , Animals , Brain/pathology , Diffusion , Mice , Myelin Basic Protein/metabolismABSTRACT
Optic nerves from mice that have undergone retinal ischemia were examined using a newly implemented quantitative magnetization transfer (qMT) technique. Previously published results indicate that the optic nerve from retinal ischemia mice suffered significant axon degeneration without detectable myelin injury at 3 days after reperfusion. At this time point, we acquired ex vivo qMT parameters from both shiverer mice (which have nearly no myelin) and control mice that have undergone retinal ischemia, and these qMT measures were compared with diffusion tensor imaging (DTI) results. Our findings suggests that the qMT estimated ratio of the pool sizes of the macromolecular and free water protons reflected the different myelin contents in the optic nerves between the shiverer and control mice. This pool size ratio was specific to myelin content only and was not significantly affected by the presence of axon injury in mouse optic nerve 3 days after retinal ischemia.
Subject(s)
Diffusion Magnetic Resonance Imaging/methods , Image Interpretation, Computer-Assisted/methods , Ischemia/pathology , Nerve Fibers, Myelinated/pathology , Optic Nerve/pathology , Retinal Artery/pathology , Animals , MiceABSTRACT
The changes of directional diffusivities derived from diffusion tensor imaging (DTI), i.e. decreased axial diffusivity (lambda(||)) and increased radial diffusivity (lambda( perpendicular)), have shown significant correlation with axonal and myelin damage, respectively. However, after formalin fixation, reduced sensitivity of lambda(||) in detecting axonal damage in tissue has raised the concern of applying DTI ex vivo. In order to distinguish whether death or the fixation process diminishes the sensitivity of DTI in detecting lesions, in vivo, pre-fixed postmortem, and fixed postmortem DTI were conducted on mouse optic nerves 3 and 14 days after transient retinal ischemia. Our data showed that, from in vivo to pre-fixed postmortem, lambda(||) and lambda( perpendicular) decreased by 50 to 70% in both healthy and injured optic nerves (3 and 14 day injury). From pre-fixed postmortem to fixed postmortem, lambda(||) and lambda( perpendicular) decreased by 40 to 50% in normal and 3-day injured optic nerves, but only by 15 to 25% in 14-day injured optic nerves. Consequently, for the 14-day injured optic nerves, the differences between healthy and injured nerves were not preserved after fixation: the 40% decreased lambda(||) and 200% increased lambda( perpendicular) in injured nerves as compared to the normal nerves were measured in vivo and pre-fixed postmortem, but after the fixation process, 300% increased lambda( perpendicular) and insignificant changes in lambda(||) were found in injured nerves as compared to the normal nerves. This study clarified that fixation process, but not death, could change the sensitivity of DTI in detecting injury.
Subject(s)
Autopsy/methods , Diffuse Axonal Injury/pathology , Diffusion Magnetic Resonance Imaging/methods , Optic Nerve Diseases/pathology , Optic Nerve Injuries/pathology , Optic Nerve/pathology , Tissue Fixation/methods , Animals , Artifacts , Image Enhancement/methods , Male , Mice , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Wallerian degeneration plays a significant role in many central nervous system (CNS) diseases. Tracking the progression of Wallerian degeneration may provide better understanding of the evolution of many CNS diseases. In this study, a 28-day longitudinal in vivo DTI of optic nerve (ON) and optic tract (OT) was conducted to evaluate the temporal and spatial evolution of Wallerian degeneration resulting from the transient retinal ischemia. At 3-28 days after ischemia, ipsilateral ON and contralateral OT showed significant reduction in axial diffusivity (32-40% and 21-29% respectively) suggestive of axonal damage. Both ON and OT showed significant increase in radial diffusivity, 200-290% and 58-65% respectively, at 9-28 days suggestive of myelin damage. Immunohistochemistry of phosphorylated neurofilament (pNF) and myelin basic protein (MBP) was performed to assess axonal and myelin integrities validating the DTI findings. Both DTI and immunohistochemistry detected that transient retinal ischemia caused more severe damage to ON than to OT. The current results suggest that axial and radial diffusivities are capable of reflecting the severity of axonal and myelin damage in mice as assessed using immunohistochemistry.
Subject(s)
Retinal Vessels/pathology , Wallerian Degeneration/pathology , Algorithms , Animals , Axons/pathology , Cell Count , Cell Death , Diffusion Magnetic Resonance Imaging , Immunohistochemistry , Male , Mice , Myelin Basic Protein/metabolism , Myelin Sheath/pathology , Neurofilament Proteins/metabolism , Optic Nerve/pathology , Phosphorylation , Retinal Ganglion Cells/pathology , Visual Pathways/pathologyABSTRACT
Recent studies have suggested that axonal damage, and not demyelination, is the primary cause of long-term neurological impairment in multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). The axial and radial diffusivities derived from diffusion tensor imaging have shown promise as non-invasive surrogate markers of axonal damage and demyelination, respectively. In this study, in vivo diffusion tensor imaging of the spinal cords from mice with chronic EAE was performed to determine if axial diffusivity correlated with neurological disability in EAE assessed by the commonly used clinical scoring system. Axial diffusivity in the ventrolateral white matter showed a significant negative correlation with EAE clinical score and was significantly lower in mice with severe EAE than in mice with moderate EAE. Furthermore, the greater decreases in axial diffusivity were associated with greater amounts of axonal damage, as confirmed by quantitative staining for non-phosphorylated neurofilaments (SMI32). Radial diffusivity and relative anisotropy could not distinguish between the groups of mice with moderate EAE and those with severe EAE. The results further the notion that axial diffusivity is a non-invasive marker of axonal damage in white matter and could provide the necessary link between pathology and neurological disability.
Subject(s)
Demyelinating Diseases/pathology , Diffuse Axonal Injury/pathology , Diffusion Magnetic Resonance Imaging/methods , Image Interpretation, Computer-Assisted/methods , Multiple Sclerosis/diagnosis , Nervous System Diseases/pathology , Spinal Cord/pathology , Animals , Disease Models, Animal , Humans , Male , Mice , Mice, Inbred C57BL , Reproducibility of Results , Sensitivity and Specificity , Statistics as TopicABSTRACT
In this study, axial (lambda(parallel)) and radial (lambda(perpendicular)) diffusivities derived from diffusion tensor imaging (DTI) were used to evaluate white matter injury in brains of mice affected by experimental autoimmune encephalomyelitis (EAE). Sixteen female C57BL/6 mice were immunized with amino acids 35-55 of myelin oligodendrocyte glycoprotein (MOG(35-55)). Three months after immunization, optic nerve and tract were severely affected with 19% and 18% decrease in lambda(parallel) respectively, suggesting the presence of axonal injury. In addition, a 156% and 86% increase in lambda( perpendicular) was observed in optic nerve and tract respectively, suggestive of myelin injury. After in vivo DTI, mice were perfusion fixed and immunohistochemistry for the identification of myelin basic protein (MBP) and phosphorylated neurofilament (pNF) was performed to verify the presence of axonal and myelin injury. The present study demonstrated that the visual pathway is selectively affected in MOG(35-55) induced murine EAE and these injuries are non-invasively detectable using lambda(parallel) and lambda( perpendicular).
Subject(s)
Diffusion Magnetic Resonance Imaging , Encephalomyelitis, Autoimmune, Experimental/pathology , Multiple Sclerosis/pathology , Optic Nerve/pathology , Visual Pathways/pathology , Animals , Female , Glycoproteins/immunology , Immunohistochemistry , Mice , Myelin-Oligodendrocyte Glycoprotein , Peptide Fragments/immunologyABSTRACT
We examined in vivo measurements of directional diffusivity derived from diffusion tensor imaging (DTI) to study the evolution of ventrolateral white matter (VWM) changes following contusive spinal cord injury (SCI) in C57BL/6 mice at 1, 3, 7, and 14 days postinjury. Relative anisotropy maps provided excellent gray matter (GM)/white matter (WM) contrast for characterization of evolving WM injury at all time points. Longitudinal DTI measurements clearly demonstrated rostral-caudal injury asymmetry. Axial diffusivity provided a sensitive, noninvasive measure of axonal integrity within the injury epicenter and at remote levels. Quantitative measurements of axial and radial diffusivities in VWM showed a trend of acute primary axonal injury followed by delayed, subacute myelin damage at the impact site, with good histological correlation.
Subject(s)
Diffusion Magnetic Resonance Imaging/methods , Spinal Cord Injuries/diagnosis , Spinal Cord Injuries/pathology , Spinal Cord/pathology , Acute Disease , Analysis of Variance , Animals , Anisotropy , Disease Models, Animal , Edema/diagnosis , Edema/etiology , Edema/physiopathology , Female , Mice , Mice, Inbred C57BL , Nerve Fibers, Myelinated/pathology , Neural Pathways/pathology , Neural Pathways/physiopathology , Predictive Value of Tests , Spinal Cord/physiopathology , Spinal Cord Injuries/physiopathologyABSTRACT
Diffusion tensor imaging (DTI) has been widely applied to investigate injuries in the central nervous system (CNS) white matter (WM). However, the underlying pathological correlates of diffusion changes have not been adequately determined. In this study the coregistration of histological sections to MR images and a pixel-based receiver operating characteristic (ROC) analysis were used to compare the axial (lambda( parallel)) and radial (lambda( perpendicular)) diffusivities derived from DTI and histological markers of axon (phosphorylated neurofilament, SMI-31) and myelin (Luxol fast blue (LFB)) integrity, respectively, in two different patterns of injury to mouse spinal cord (SC) WM. In contusion SC injury (SCI), a decrease in lambda( parallel) matched the pattern of axonal damage with high accuracy, but lambda( perpendicular) did not match the pattern of demyelination detected by LFB. In a mouse model of multiple sclerosis (MS), lambda( perpendicular) and lambda( parallel) did not match the patterns of demyelination or axonal damage, respectively. However, a region of interest (ROI) analysis suggested that lambda( perpendicular)-detected demyelination paralleled that observed with LFB, and lambda( parallel) decreased in both regions of axonal damage and normal-appearing WM (NAWM) as visualized by SMI-31. The results suggest that directional diffusivities may reveal abnormalities that are not obvious with SMI-31 and LFB staining, depending on the type of injury.
Subject(s)
Demyelinating Autoimmune Diseases, CNS/diagnosis , Diffusion Magnetic Resonance Imaging/methods , Nerve Fibers, Myelinated/pathology , Animals , Demyelinating Autoimmune Diseases, CNS/pathology , Diffuse Axonal Injury/diagnosis , Diffuse Axonal Injury/pathology , Disease Models, Animal , Male , Mice , Mice, Inbred C57BL , Multiple Sclerosis/diagnosis , Multiple Sclerosis/pathology , ROC Curve , Statistics, NonparametricABSTRACT
Decreased axial (lambda(||)) and increased radial (lambda( perpendicular)) diffusivity have been shown to reflect axonal and myelin injury respectively. In the present study, evolving white matter injury within the optic nerves of mice with retinal ischemia was examined by in vivo and ex vivo measurements of lambda(||) and lambda( perpendicular). The results show that at 3 days after retinal ischemia, a 33% decrease in vivo and a 38% decrease ex vivo in lambda(||) without change in lambda( perpendicular) was observed in the injured optic nerve compared to the control, suggestive of axonal damage without myelin injury. At 14 days, both in vivo and ex vivo measured lambda( perpendicular) increased significantly to 220-240% of the control level in the injured optic nerve suggestive of myelin damage. In contrast, the axonal injury that was clearly detected in vivo as a significantly decreased lambda(||) (33% decrease) was not as clearly detected by ex vivo lambda(||) (17% decrease). The current findings suggest that ex vivo lambda( perpendicular) is comparable to in vivo lambda( perpendicular) in detecting myelin injury. However, the structural changes resulting from axonal damage causing the decreased in vivo lambda(||) may not be preserved ex vivo in the fixed tissues. Despite the accurate depiction of the pathology using lambda(||) and lambda( perpendicular) in vivo, the use of ex vivo lambda(||) to extrapolate the status of axonal injury in vivo would require further investigation.
Subject(s)
Optic Nerve Diseases/pathology , Optic Neuropathy, Ischemic/pathology , Algorithms , Animals , Axons/pathology , Brain/pathology , Data Interpretation, Statistical , Diffusion Magnetic Resonance Imaging , Immunohistochemistry , Male , Mice , Myelin Sheath/pathology , Nerve Degeneration/pathology , Neurofilament Proteins/metabolism , Optic Nerve Diseases/etiology , Optic Neuropathy, Ischemic/complications , Tissue FixationABSTRACT
Previously, we tested the prediction that axonal damage results in decreased axial diffusivity (lambda(parallel)) while demyelination leads to increased radial diffusivity (lambda(perpendicular)). Cuprizone treatment of C57BL/6 mice was a highly reproducible model of CNS white matter demyelination and remyelination affecting the corpus callosum (CC). In the present study, six C57BL/6 male mice were fed 0.2% cuprizone for 12 weeks followed by 12 weeks of recovery on normal chow. The control mice were fed normal chow and imaged in parallel. Biweekly in vivo DTI examinations showed transient decrease of lambda(parallel) in CC at 2-6 weeks of cuprizone treatment. Immunostaining for nonphosphorylated neurofilaments demonstrated corresponding axonal damage at 4 weeks of treatment. Significant demyelination was evident from loss of Luxol fast blue staining at 6-12 weeks of cuprizone ingestion and was paralleled by increased lambda(perpendicular) values, followed by partial normalization during the remyelination phase. The sensitivity of lambda(perpendicular) to detect demyelination may be modulated in the presence of axonal damage during the early stage of demyelination at 4 weeks of cuprizone treatment. Our results suggest that lambda(parallel) and lambda(perpendicular) may be useful in vivo surrogate markers of axonal and myelin damage in mouse CNS white matter.