Diffusion kurtosis imaging probes cortical alterations and white matter pathology following cuprizone induced demyelination and spontaneous remyelination.

Although MRI is the gold standard for the diagnosis and monitoring of multiple sclerosis (MS), current conventional MRI techniques often fail to detect cortical alterations and provide little information about gliosis, axonal damage and myelin status of lesioned areas. Diffusion tensor imaging (DTI) and diffusion kurtosis imaging (DKI) provide sensitive and complementary measures of the neural tissue microstructure. Additionally, specific white matter tract integrity (WMTI) metrics modelling the diffusion in white matter were recently derived. In the current study we used the well-characterized cuprizone mouse model of central nervous system demyelination to assess the temporal evolution of diffusion tensor (DT), diffusion kurtosis tensor (DK) and WMTI-derived metrics following acute inflammatory demyelination and spontaneous remyelination. While DT-derived metrics were unable to detect cuprizone induced cortical alterations, the mean kurtosis (MK) and radial kurtosis (RK) were found decreased under cuprizone administration, as compared to age-matched controls, in both the motor and somatosensory cortices. The MK remained decreased in the motor cortices at the end of the recovery period, reflecting long lasting impairment of myelination. In white matter, DT, DK and WMTI-derived metrics enabled the detection of cuprizone induced changes differentially according to the stage and the severity of the lesion. More specifically, the MK, the RK and the axonal water fraction (AWF) were the most sensitive for the detection of cuprizone induced changes in the genu of the corpus callosum, a region less affected by cuprizone administration. Additionally, microgliosis was associated with an increase of MK and RK during the acute inflammatory demyelination phase. In regions undergoing severe demyelination, namely the body and splenium of the corpus callosum, DT-derived metrics, notably the mean diffusion (MD) and radial diffusion (RD), were among the best discriminators between cuprizone and control groups, hence highlighting their ability to detect both acute and long lasting changes. Interestingly, WMTI-derived metrics showed the aptitude to distinguish between the different stages of the disease. Both the intra-axonal diffusivity (Da) and the AWF were found to be decreased in the cuprizone treated group, Da specifically decreased during the acute inflammatory demyelinating phase whereas the AWF decrease was associated to the spontaneous remyelination and the recovery period. Altogether our results demonstrate that DKI is sensitive to alterations of cortical areas and provides, along with WMTI metrics, information that is complementary to DT-derived metrics for the characterization of demyelination in both white and grey matter and subsequent inflammatory processes associated with a demyelinating event.

Fetal organ weight estimation by postmortem high-field magnetic resonance imaging before 20 weeks' gestation.

OBJECTIVE: To ascertain whether high-field magnetic resonance imaging (MRI) allows accurate estimation of the weight of various fetal organs at postmortem before 20 weeks' gestation. METHODS: From 23 fetuses at 9-20 weeks, following termination of pregnancy or in-utero fetal death (IUFD), 207 assorted fetal organs were evaluated by high-field MRI at 9.4 T prior to conventional autopsy. Fetal organ density was calculated by correlating volume and weight at autopsy using linear regression analysis, and this was used to estimate fetal organ weight by MRI. The relative error in MRI estimation of organ weight was calculated as follows: (|MRI weight - autopsy weight|/autopsy weight) × 100 (%). Multiple regression analysis was used to investigate the effect on the relative error of MRI organ weight estimates of gestational age at TOP or delivery following IUFD, autopsy weight, fetal organ examined, IUFD and fetal maceration. RESULTS: Of the 207 organs evaluated, 133 (64%) were examined for fetal organ density and 155 (75%) for fetal organ weight. Fifty-two organs were excluded from our analysis; 41 of these were from fetuses with IUFD, with 39 organs macerated. In 32 cases, exclusion was due to an inability to assess the organ both on MRI and on conventional autopsy. Volume and weight at autopsy correlated significantly, following the linear equation: autopsy volume = (0.9947 × autopsy weight) - 4.7556, where autopsy volume is in mm(3) and weight is in mg (r = 0.99, P < 0.001). Overall the mean relative error in the MRI estimation of organ weight was 68%. Multiple regression analysis showed that the relative error in the MRI estimation of organ weight was significantly associated with gestational age at TOP or delivery following IUFD and fetal maceration, but not with autopsy weight, fetal organ examined or IUFD. In the subgroup of non-macerated organs and for fetuses above 14 weeks' gestation, the mean relative error in the MRI estimation of organ weight was 34%. CONCLUSION: In fetuses before 20 weeks' gestation, noninvasive estimation of organ weight is feasible using high-field MRI, but there is a mean overestimation. Limitations of the technique occur mainly in cases of small macerated fetuses before 14 weeks' gestation.

Postmortem examination of human fetal hearts at or below 20 weeks' gestation: a comparison of high-field MRI at 9.4 T with lower-field MRI magnets and stereomicroscopic autopsy.

OBJECTIVES: To compare the diagnostic usefulness of high-field with low-field magnetic resonance imaging (MRI) and stereomicroscopic autopsy for examination of the heart in fetuses at or under 20 weeks' gestation. METHODS: Prior to invasive stereomicroscopic autopsy, MRI scans at 9.4, 3.0 and 1.5 T were performed on 24 fetuses between 11 and 20 weeks' gestation, including 10 fetuses with cardiac abnormalities. The ability to visualize different heart structures was evaluated according to the different field strength MRI magnets used and gestational age at examination. RESULTS: On 1.5- and 3.0-T MRI, only the heart situs and four-chamber view could be visualized consistently (in 75% or more of cases) when the fetus was beyond 16 weeks' gestation, but other heart structures could not be visualized for fetuses at any gestational age. In contrast, using high-field MRI at 9.4 T, the heart situs, four-chamber view and the outflow tracts could be visualized in all fetuses irrespective of gestational age. Using high-field MRI, the sensitivity for detecting an abnormality of the four-chamber view was 66.7% (95% CI, 30.1-92.1%) with a specificity of 80.0% (95% CI, 51.9-95.4%). For abnormalities of the outflow tracts, sensitivity was 75.0% (95% CI, 20.3-95.9%) and specificity 100.0% (95% CI, 83.3-100.0%). Eight fetuses out of 10 with congenital heart disease (CHD) were classified as having major CHD. High-field MRI at 9.4 T was able to identify seven out of the eight cases of major CHD. CONCLUSION: High-field MRI at 9.4 T seems to be an acceptable alternative approach to invasive stereomicroscopic autopsy for fetuses with CHD at or below 20 weeks' gestation.

Measuring brain hemodynamic changes in a songbird: responses to hypercapnia measured with functional MRI and near-infrared spectroscopy.

Songbirds have been evolved into models of choice for the study of the cerebral underpinnings of vocal communication. Nevertheless, there is still a need for in vivo methods allowing the real-time monitoring of brain activity. Functional Magnetic Resonance Imaging (fMRI) has been applied in anesthetized intact songbirds. It relies on blood oxygen level-dependent (BOLD) contrast revealing hemodynamic changes. Non-invasive near-infrared spectroscopy (NIRS) is based on the weak absorption of near-infrared light by biological tissues. Time-resolved femtosecond white laser NIRS is a new probing method using real-time spectral measurements which give access to the local variation of absorbing chromophores such as hemoglobins. In this study, we test the efficiency of our time-resolved NIRS device in monitoring physiological hemodynamic brain responses in a songbird, the zebra finch (Taeniopygia guttata), using a hypercapnia event (7% inhaled CO(2)). The results are compared to those obtained using BOLD fMRI. The NIRS measurements clearly demonstrate that during hypercapnia the blood oxygen saturation level increases (increase in local concentration of oxyhemoglobin, decrease in deoxyhemoglobin concentration and total hemoglobin concentration). Our results provide the first correlation in songbirds of the variations in total hemoglobin and oxygen saturation level obtained from NIRS with local BOLD signal variations.

Auditory evoked BOLD responses in awake compared to lightly anaesthetized zebra finches.

Functional magnetic resonance imaging (fMRI) is increasingly used in cognitive neuroscience and has become a valuable tool in the study of auditory processing in zebra finches, a well-established model of learned vocal communication. Due to its sensitivity to head motion, most fMRI studies in animals are performed in anaesthetized conditions, which might significantly impact neural activity evoked by stimuli and cognitive tasks. In this study, we (1) demonstrate the feasibility of fMRI in awake zebra finches and (2) explore how light anaesthesia regimes affect auditory-evoked BOLD responses to biologically relevant songs. After an acclimation procedure, we show that fMRI can be successfully performed during wakefulness, enabling the detection of reproducible BOLD responses to sound. Additionally, two light anaesthesia protocols were tested (isoflurane and a combination of medetomidine and isoflurane), of which isoflurane alone appeared to be the most promising given the high success rate, non-invasive induction, and quick recovery. By comparing auditory evoked BOLD responses in awake versus lightly anaesthetized conditions, we observed overall effects of anaesthetics on cerebrovascular reactivity as reflected in the extent of positive and negative BOLD responses. Further, our results indicate that light anaesthesia has limited effects on selective BOLD responses to natural versus synthetic sounds.

Fibrillin-1 impairment enhances blood-brain barrier permeability and xanthoma formation in brains of apolipoprotein E-deficient mice.

We recently reported that apolipoprotein E (ApoE)-deficient mice with a mutation in the fibrillin-1 gene (ApoE(-/-)Fbn1(C1039G+/-)) develop accelerated atherosclerosis with enhanced inflammation, atherosclerotic plaque rupture, myocardial infarction and sudden death. In the brain, fibrillin-1 functions as an attachment protein in the basement membrane, providing structural support to the blood-brain barrier (BBB). Here, we investigated whether fibrillin-1 impairment affects the permeability of the BBB proper and the blood-cerebrospinal fluid barrier (BCSFB), and whether this leads to the accelerated accumulation of lipids (xanthomas) in the brain. ApoE(-/-) (n=61) and ApoE(-/-)Fbn1(C1039G+/-) (n=73) mice were fed a Western-type diet (WD). After 14 weeks WD, a significantly higher permeability of the BBB was observed in ApoE(-/-)Fbn1(C1039G+/-) mice compared to age-matched ApoE(-/-) mice. This was accompanied by leukocyte infiltration, enhanced expression of pro-inflammatory cytokines, matrix metalloproteinases and transforming growth factor-ß, and by decreased expression of tight junction proteins claudin-5 and occludin. After 20 weeks WD, 83% of ApoE(-/-)Fbn1(C1039G+/-) mice showed xanthomas in the brain, compared to 23% of their ApoE(-/-) littermates. Xanthomas were mainly located in fibrillin-1-rich regions, such as the choroid plexus and the neocortex. Our findings demonstrate that dysfunctional fibrillin-1 impairs BBB/BCSFB integrity, facilitating peripheral leukocyte infiltration, which further degrades the BBB/BCSFB. As a consequence, lipoproteins can enter the brain, resulting in accelerated formation of xanthomas.

Brain studies of mouse models for neurogenetic disorders using in vivo magnetic resonance imaging (MRI).

Magnetic resonance imaging (MRI) is a technique commonly used to detect neural abnormalities in routine clinical practice. It is perhaps less well known that the technique can be adapted to measure various anatomical and physiological features of small laboratory rodents. This review focuses on the potential of the MRI technique to image the brain of (transgenic) mouse models for neurological diseases, and aims to introduce these exciting new technological developments to the non-specialist reader.

Neuroanatomy of the fragile X knockout mouse brain studied using in vivo high resolution magnetic resonance imaging.

Magnetic resonance imaging (MRI) of the brain of fragile X patients, the most frequent form of inherited mental retardation, has revealed abnormalities in the size of specific brain structures, including the cerebellar vermis, the hippocampus, and the ventricular system. We intended to quantify the differences observed in the patient studies in the fragile X knockout mouse model, which is a good model for the disease, paralleling the human disorder in having cognitive deficits, macro-orchidism, and immature dendritic spines. Therefore we set up MRI of the mouse brain which allowed us to measure the size of the brain structures reported to be abnormal in human fragile X patients in the mouse model. We did not find evidence for size alterations in various brain regions of the fragile X mouse model, but the method described may find a wide application in the study of mutant mouse models with neurological involvement.

Diffusion-weighted MRI of infarct growth in a rat photochemical stroke model: effect of lubeluzole.

We studied the neuroprotective effect of lubeluzole, a NOS (nitric oxide synthase) pathway modulator, on the development of ischemic damage within the first six hours after a photochemically induced neocortical infarct in rats using diffusion-weighted MRI and Apparent Diffusion Coefficient (ADC) maps. A unilateral photochemical infarct was induced in the hindlimb sensorimotor neocortex of Wistar rats. One hour after infarction, rats received either vehicle (n=10) or lubeluzole (n=11; a 0.31 mg/kg i.v. bolus followed by a one-hour 0.31 mg/kg i.v. infusion). During the first six hours after infarct induction, multislice T2- and Diffusion-Weighted magnetic resonance images (MRI) were obtained to measure percent change of volume of ischemic damage, whereas regional ADC maps were used to measure time-dependent density of ischemic damage. Lubeluzole reduced the percent increase of volume of ischemic damage relative to baseline (at 1 h after infarct induction just before drug treatment), by 18% at 5 and 6 hrs after infarct induction. Lubeluzole attenuated the ADC decreases in the peripheral rim of the infarct, but left the ADC values in the core unaffected. In conclusion, the neuroprotectant lubeluzole attenuates growth of ischemic damage as well as its density in the periphery of a photochemically induced neocortical infarct in rats.

In vivo manganese-enhanced magnetic resonance imaging reveals connections and functional properties of the songbird vocal control system.

Injection of manganese (Mn(2+)), a paramagnetic tract tracing agent and calcium analogue, into the high vocal center of starlings labeled within a few hours the nucleus robustus archistriatalis and area X as observed by in vivo magnetic resonance imaging. Structures highlighted by Mn(2+) accumulation assumed the expected tri-dimensional shape of the nucleus robustus archistriatalis and area X as identified by classical histological or neurochemical methods. The volume of these nuclei could be accurately calculated by segmentation of the areas highlighted by Mn(2+). Besides confirming previously established volumetric sex differences, Mn(2+) uptake into these nuclei revealed new functional sex differences affecting Mn(2+) transport. A faster transport was observed in males than in females and different relative amounts of Mn(2+) were transported to nucleus robustus archistriatalis and area X in males as compared to females. This new in vivo approach, allowing repeated measures, opens new vistas to study the remarkable seasonal plasticity in size and activity of song-control nuclei and correlate neuronal activity with behavior. It also provides new insights on in vivo axonal transport and neuronal activity in song-control nuclei of oscines.

Neonatal neuronal overexpression of glycogen synthase kinase-3 beta reduces brain size in transgenic mice.

Glycogen synthase kinase-3beta (GSK-3beta) is important in neurogenesis. Here we demonstrate that the kinase influenced post-natal maturation and differentiation of neurons in vivo in transgenic mice that overexpress a constitutively active GSK-3beta[S9A]. Magnetic resonance imaging revealed a reduced volume of the entire brain, concordant with a nearly 20% reduction in wet brain weight. The reduced volume was most prominent for the cerebral cortex, without however, disturbing the normal cortical layering. The resulting compacted architecture was further demonstrated by an increased neuronal density, by reduced size of neuronal cell bodies and of the somatodendritic compartment of pyramidal neurons in the cortex. No evidence for apoptosis was obtained. The marked overall reduction in the level of the microtubule-associated protein 2 in brain and in spinal cord, did not affect the ultrastructure of the microtubular cytoskeleton in the proximal apical dendrites. The overall reduction in size of the entire CNS induced by constitutive active GSK-3beta caused only very subtle changes in the psychomotoric ability of adult and ageing GSK-3beta transgenic mice.

Non invasive in vivo anatomical studies of the oscine brain by high resolution MRI microscopy.

We describe in this paper an in vivo Magnetic Resonance Imaging (MRI) procedure that allows one to obtain three-dimensional high quality images of the entire brain of small passerine birds such as the canary with a slice thickness of 58 micron and an image resolution of 78 microns. This imaging procedure was completed in 70 min on anaesthetised birds that later recovered uneventfully and could be reused for subsequent additional imaging. To illustrate the high resolution and anatomical detail that can be achieved, examples of coronal images through the entire hypothalamus are provided in the same sectioning plane as the previously published canary brain atlas. The data set can be used to create sections in any desired plane and the entire data set can be viewed from any point of view in a volume rendered image. This provides a useful tool in understanding the three-dimensional organisation of the brain. Similar procedures can also be applied on fixed brains and might allow an even better anatomical resolution of images because time constrains no longer limit the duration of image acquisition. The non-invasive MRI technique enables to study neuroanatomical features with a high resolution and without killing the animal subjects so that measures can be obtained in a same individual both before and after an experimental treatment.

Watershed-based segmentation of 3D MR data for volume quantization.

The aim of this work is the development of a semiautomatic segmentation technique for efficient and accurate volume quantization of Magnetic Resonance (MR) data. The proposed technique uses a 3D variant of Vincent and Soilles immersion-based watershed algorithm that is applied to the gradient magnitude of the MR data and that produces small volume primitives. The known drawback of the watershed algorithm, oversegmentation, is strongly reduced by a priori application of a 3D adaptive anisotropic diffusion filter to the MR data. Furthermore, oversegmentation is a posteriori reduced by properly merging small volume primitives that have similar gray level distributions. The outcome of the proceeding image processing steps is presented to the user for manual segmentation. Through selection of volume primitives, the user quickly segments of first slice, which contains the object of interest. Afterwards, the subsequent slices are automatically segmented by extrapolation. Segmentation results are contingently manually corrected. The proposed segmentation technique is tested on phantom objects, where segmentation errors less than 2% are observed. In addition, the technique is demonstrated on 3D MR data of the mouse head from which the cerebellum is extracted. Volumes of the mouse cerebellum and the mouse brains in toto are calculated.

In vivo noninvasive determination of abnormal water diffusion in the rat brain studied in an animal model for multiple sclerosis by diffusion-weighted NMR imaging.

In vivo NMR images of the rat brain were obtained using a NMR microscope (7 T) from SMIS (England). Four animals were imaged every 3-4 days during a pathological cycle (starting after induction and up to 37 days) of experimental allergic encephalomyelitis (EAE), an animal model for multiple sclerosis. The EAE rats were weighted and clinically scored daily. We aimed at measuring the apparent diffusion coefficient (ADC) or the mean diffusivity (D) with a high accuracy, and within a reasonable experimental time frame, because of the clinical situation of the animals. Therefore, we fitted the ADC value from five diffusion-weighted images--with an experimental time of 17 min/image--and chose to apply diffusion-sensitizing gradients in a direction intersecting all fiber directions of the external capsule. With this, we also obtained high b-values. For the control rats, we obtained a statistical mean value of ADC = (388 +/- 16) 10(-12) m2/s for gray matter and a statistical mean value of (D) of (750 +/- 30) 10(-12) m2/s for white matter, measured in the external capsule. For the EAE rats, no alterations in ADC values of gray matter with increasing clinical scores were observed. Concerning white matter, as determined in the external capsule, there were no significant differences in (D) values between controls and EAE rats before clinical signs occurred. However, when clinical signs were observed, we could demonstrate a significant positive correlation between the clinical score and the (D) values in the external capsule. As the clinical signs became more severe, we measured a rise in water diffusion (increase in (D)) in the external capsule, which was accompanied by the occurrence of interstitial edema as revealed by a complementary histological study.

Reduction of ECG and gradient related artifacts in simultaneously recorded human EEG/MRI data.

Nowadays, electroencephalography signals can be acquired from a patient lying in a magnetic resonance imaging system. It is even possible to acquire EEG signals during an MR imaging sequence. However, such EEG signals are severely distorted by artifacts originating from various effects (e.g., MR gradients, ECG). In this paper, a simple method is presented to reduce such artifacts. Thereby, special attention is focused on artifacts related to the patient's electrocardiogram. The method is shown to be effective, adaptive, and automatic.

Special designed RF-antenna with integrated non-invasive carbon electrodes for simultaneous magnetic resonance imaging and electroencephalography acquisition at 7T.

The construction of a high quality MR RF-antenna with incorporated EEG electrodes for simultaneous MRI and EEG acquisition is presented. The antenna comprises an active decoupled surface coil for receiving the MR signal and a whole body coil for transmitting the excitation RF pulses. The surface coil offers a high signal-to-noise ratio required for fMRI application and the whole body coil has a good B(1) excitation profile, which enables the application of homogeneous RF pulses. Non-invasive carbon electrodes are used in order to minimise the magnetic susceptibility artefacts that occur upon application of conductive materials. This dedicated set-up is compared to a standard set-up being a linear birdcage coil and commercially available Ag/AgCl electrodes. As the acquired EEG signals are heavily disturbed by the gradient switching, intelligent filtering is applied to obtain a clean EEG signal.

Restoration of MR-induced artifacts in simultaneously recorded MR/EEG data.

During a Magnetic Resonance sequence, simultaneously acquired ElectroEncephaloGraphy (EEG) data are compromised by severe pollution due to artifacts originating from the switching of the magnetic field gradients. In this work, it is shown how these artifacts can be strongly reduced or even removed through application of an adaptive artifact restoration scheme. The method has proved to be fully automatic and to retain high frequency EEG information, which is indispensable for many EEG applications.

Estimation of the noise in magnitude MR images.

Magnitude magnetic resonance data are Rician distributed. In this note a new method is proposed to estimate the image noise variance for this type of data distribution. The method is based on a double image acquisition, thereby exploiting the knowledge of the Rice distribution moments.

Automatic localization of EEG electrode markers within 3D MR data.

The electrical activity of the brain can be monitored using ElectroEncephaloGraphy (EEG). From the positions of the EEG electrodes, it is possible to localize focal brain activity. Thereby, the accuracy of the localization strongly depends on the accuracy with which the positions of the electrodes can be determined. In this work, we present an automatic, simple, and accurate scheme that detects EEG electrode markers from 3D MR data of the human head.

Subchronic memantine induced concurrent functional disconnectivity and altered ultra-structural tissue integrity in the rodent brain: revealed by multimodal MRI.

BACKGROUND: An effective NMDA antagonist imaging model may find key utility in advancing schizophrenia drug discovery research. We investigated effects of subchronic treatment with the NMDA antagonist memantine by using behavioural observation and multimodal MRI. METHODS: Pharmacological MRI (phMRI) was used to map the neuroanatomical binding sites of memantine after acute and subchronic treatment. Resting state fMRI (rs-fMRI) and diffusion MRI were used to study the changes in functional connectivity (FC) and ultra-structural tissue integrity before and after subchronic memantine treatment. Further corroborating behavioural evidences were documented. RESULTS: Dose-dependent phMRI activation was observed in the prelimbic cortex following acute doses of memantine. Subchronic treatment revealed significant effects in the hippocampus, cingulate, prelimbic and retrosplenial cortices. Decreases in FC amongst the hippocampal and frontal cortical structures (prelimbic, cingulate) were apparent through rs-fMRI investigation, indicating a loss of connectivity. Diffusion kurtosis MRI showed decreases in fractional anisotropy and mean diffusivity changes, suggesting ultra-structural changes in the hippocampus and cingulate cortex. Limited behavioural assessment suggested that memantine induced behavioural effects comparable to other NMDA antagonists as measured by locomotor hyperactivity and that the effects could be reversed by antipsychotic drugs. CONCLUSION: Our findings substantiate the hypothesis that repeated NMDA receptor blockade with nonspecific, noncompetitive NMDA antagonists may lead to functional and ultra-structural alterations, particularly in the hippocampus and cingulate cortex. These changes may underlie the behavioural effects. Furthermore, the present findings underscore the utility and the translational potential of multimodal MR imaging and acute/subchronic memantine model in the search for novel disease-modifying treatments for schizophrenia.