Galit Pelled

Detection of cortical laminar architecture using manganese-enhanced MRI.

Changes in manganese-enhanced MRI (MEMRI) contrast across the rodent somatosensory cortex were compared to the cortical laminae as identified by tissue histology and administration of an anatomical tracer to cortex and thalamus. Across the cortical thickness, MEMRI signal intensity was low in layer I, increased in layer II, decreased in layer III until mid-layer IV, and increased again, peaking in layer V, before decreasing through layer VI. The reeler mouse mutant was used to confirm that the cortical alternation in MEMRI contrast was related to laminar architecture. Unlike in wild-type mice, the reeler cortex showed no appreciable changes in MEMRI signal, consistent with absence of cortical laminae in histological slides. The tract tracing ability of MEMRI was used to further confirm assignments and demonstrate laminar specificity. Twelve to 16 h after stereotaxic injections of MnCl(2) to the ventroposterior thalamic nuclei, an overall increase in signal intensity was detected in primary somatosensory cortex compared to other brain regions. Maximum intensity projection images revealed a distinctly bright stripe located 600-700 microm below the pial surface, in layer IV. The data show that both systemic and tract tracing forms of MEMRI are useful for studying laminar architecture in the brain.

Bilateral overactivation of the sensorimotor cortex in the unilateral rodent model of Parkinson's disease: a functional magnetic resonance imaging study

Functional magnetic resonance imaging (fMRI) is used to investigate the basal ganglia (BG)–cortex circuit using a rat model of Parkinsons disease (PD). The model involves a unilateral destruction of the right substantia nigra by intranigral injection of the dopaminergic neurotoxin 6‐hydroxydopamine. Volume of cortical activity was measured by the blood oxygenation level‐dependent contrast method while applying electrical forepaw stimulation. The main findings are the following. (i) Contrary to the predictions of the classic model but in line with recent experimental results (positron emission tomography, fMRI and electrophysiology), an increased cortical activity in the sensorimotor cortex of PD rats compared with sham‐operated or normal rats was found. (ii) A diffuse neuronal activity at large cortical areas that were not related directly to the stimulation used, was observed. (iii) No difference was found between the lesion and the nonlesion hemispheres when the left or the right forepaw was stimulated; both cortices show significant overactivation of the sensorimotor cortices in addition to diffuse cortical activation. The last finding could be explained by either corticocortical connections or by bilateral BG–cortex connections. These finding suggest that the mutual influence of the two hemispheres is important in the pathophysiology of the BG–cortex circuit and might be crucial in predicting treatments.

Layer specific tracing of corticocortical and thalamocortical connectivity in the rodent using manganese enhanced MRI

Information about layer specific connections in the brain comes mainly from classical neuronal tracers that rely on histology. Manganese Enhanced MRI (MEMRI) has mapped connectivity along a number of brain pathways in several animal models. It is not clear at what level of specificity neuronal connectivity measured using MEMRI tracing can resolve. The goal of this work was to determine if neural tracing using MEMRI could distinguish layer inputs of major pathways of the cortex. To accomplish this, tracing was performed between hemispheres of the somatosensory (S1) cortex and between the thalamus and S1 cortex. T(1) mapping and T(1) weighted pulse sequences detected layer specific tracing after local injection of MnCl(2). Approximately 12 h following injections into S1 cortex, maximal T(1) reductions were observed at 0.6+/-0.07 and 1.1+/-0.12 mm from the brain surface in the contralateral S1. These distances correspond to the positions of layer 3 and 5 consistent with the known callosal inputs along this pathway. Four to six hours following injection of MnCl(2) into the thalamus there were maximal T(1) reductions between 0.7+/-0.08 and 0.8+/-0.08 mm from the surface of the brain, which corresponds to layer 4. This is consistent with terminations of the known thalamocortical projections. In order to observe the first synapse projection, it was critical to perform MRI at the right time after injections to detect layer specificity with MEMRI. At later time points, tracing through the cortical network led to more uniform contrast throughout the cortex due to its complex neuronal connections. These results are consistent with well established neuronal pathways within the somatosensory cortex and demonstrate that layer specific somatosensory connections can be detected in vivo using MEMRI.

Manganese-enhanced MRI in a rat model of Parkinson's disease

To measure intra‐ and inter‐hemispheric connectivity within the basal ganglia (BG) nuclei in healthy and in unilateral 6‐hydroxydopamine (6‐OHDA) Parkinson disease rat model in order to test the BG interhemispheric connectivity hypothesis.

Ipsilateral cortical fMRI responses after peripheral nerve damage in rats reflect increased interneuron activity

In the weeks following unilateral peripheral nerve injury, the deprived primary somatosensory cortex (SI) responds to stimulation of the ipsilateral intact limb as demonstrated by functional magnetic resonance imaging (fMRI) responses. The neuronal basis of these responses was studied by using high-resolution fMRI, in vivo electrophysiological recordings, and juxtacellular neuronal labeling in rats that underwent an excision of the forepaw radial, median, and ulnar nerves. These nerves were exposed but not severed in control rats. Significant bilateral increases of fMRI responses in SI were observed in denervated rats. In the healthy SI of the denervated rats, increases in fMRI responses were concordant with increases in local field potential (LFP) amplitude and an increased incidence of single units responding compared with control rats. In contrast, in the deprived SI, increases in fMRI responses were associated with a minimal change in LFP amplitude but with increased incidence of single units responding. Based on action potential duration, juxtacellular labeling, and immunostaining results, neurons responding to intact forepaw stimulation in the deprived cortex were identified as interneurons. These results suggest that the increases in fMRI responses in the deprived cortex reflect increased interneuron activity.

Different Physiological MRI Noise Between Cortical Layers

Significantly higher temporal fluctuations of the blood oxygenation level‐dependent (BOLD) signal in the living rat group compared to that in the dead rat group were observed in the cortex, suggesting the existence of physiological information in the signal fluctuations. A similar analysis shows significantly different fluctuations between visual cortical layers. The highest fluctuations were observed in layers 4 and 5 and the lowest in layer 1. Given the consistency with published electrophysiology studies anticipating high spontaneous activity in the deeper layers (particularly layer 4), and low activity in superficial layers, we hypothesize that the BOLD signal temporal fluctuations may reflect cortical neuronal activity. Temporal fluctuations in ultrahigh spatial resolution data of the rat brain were measured in two ways. In the first, analyses were performed according to known layer widths, and in the second equal lines of 117 μ along the cortex were selected. The second approach yielded temporal fluctuations along the cortex that resemble known neuronal density distributions including the intralayer structure, particularly within layer 5. Magn Reson Med 52:913–916, 2004.

Reduced basal activity and increased functional homogeneity in sensorimotor and striatum of a Parkinson's disease rat model: a functional MRI study

Functional neuro‐imaging studies of Parkinsons disease (PD) patients and animal models show inconsistent cortical responses to sensory stimulation: some present increased sensorimotor cortex activation contradicting classical basal ganglia–cortex circuitry models, whereas others show decreased activation. As functional neuro‐imaging activation is defined as the signal difference between stimulation ON and stimulation OFF, reduced ‘activation’ can point to either increased neuronal activity during stimulation ON or to decreased basal neuronal activity during stimulation OFF. A unique non‐invasive method that uses the temporal and the spatial variances of functional magnetic resonance imaging signal is employed here to compare basal neuronal activity levels and ‘functional homogeneity’ between groups. Based on the assumption that the temporal variance reflects average neuronal activity, the variance of activity within a predefined region is defined as the regions ‘functional homogeneity’, which is assumed to estimate neuronal synchronization. Comparison of temporal and spatial variances of the sensorimotor cortex and the striatum in the 6‐hydroxydopamine (6‐OHDA) PD rat model and a control rat group show bilaterally decreased temporal and spatial variances in the 6‐OHDA rat group, suggesting bilateral reduction of basal neuronal activity levels together with an increase in local neuronal synchronization in line with classical basal ganglia–cortex circuit models.

Catheter confocal fluorescence imaging and functional magnetic resonance imaging of local and systems level recovery in the regenerating rodent sciatic nerve

The goal of the present work was to develop minimally invasive imaging techniques to monitor local regeneration of peripheral nerves and to determine the extent of return to function of brain cortical regions associated with that nerve. The sciatic nerve crush model was applied to Sprague-Dawley rats and conventional histological staining for myelin, axons and cell architecture was carried out, as well as traditional behavioral testing, to verify that nerve regeneration was occurring. The rate of sciatic nerve regeneration was measured by determining the distance a lipophilic, fluorescence probe (DiO) would move along the nerves membrane following a direct injection into the sciatic nerve. This movement was monitored using a catheter based, confocal fluorescence microscope. Two to five days after the crush, the dye moved 1.4 + 0.6 mm/day, as compared to a distance of 5.3 + 0.5 mm/day in the normal nerve. Between 9 and 13 days following the crush, the distance the dye moved increases to 5.5 + 0.5 mm/day, similar to the control, and by 15 days following the crush, the distance increased to 6.5 + 0.9 mm/day. Functional Magnetic Resonance Imaging (fMRI) measurements were performed on alpha-chloralose anesthetized rats to monitor the return of somatosensory cortical functions, which were activated by the stimulation of the lesioned peripheral nerve. fMRI results showed the return of cortical activation around 15 days following the crush procedure. However, the somatosensory cortical region activated by stimulating the crushed hindpaw was significantly smaller in extent than the intact hindpaw stimulation. These findings demonstrate that fluorescence imaging and fMRI can integrate local and system level correlates of nerve regeneration in a non-destructive manner, thus enabling serial imaging of individual animals.

Tracking the effects of crusher gradients on gradient-echo BOLD signal in space and time during rat sensory stimulation

A unique method to map the effect of crusher gradients in space and time on the gradient echo blood oxygen level dependent (BOLD) signal is introduced. Using the Radial Correlation Contrast (RCC) analysis method, amplitude‐RCC maps at different time segments and different gradient strengths were obtained. The ratio of amplitude‐RCC cluster volumes, with and without crusher gradients, showed a temporal dependency with stronger volume reduction for stimulation‐onset versus stimulation‐decline. Aside from signal‐to‐noise ratio reduction in diffusion weighted images, the average temporal patterns were equal. Comparison of the data with and without crushers showed a stronger reduction in local coherence for stimulation‐onset times. We hypothesize that the stimulation decline was weighted by extravascular effects originating in expanded veins due to their larger volume and long range susceptibility which couples neighboring voxels. The ratio of amplitude‐RCC with and without crushers calculated for each voxel at each time segment yielded a spatial–temporal mapping of the crusher effect. These maps suggest that early stimulation‐onset (∼9 s) is weighted by flow; later a dynamic steady‐state between intra‐ and extravascular effects is obtained. Stimulation‐decline was dominated by extravascular effects, and at late stimulation decline as well as at early stimulation onset, clusters were small and localized to expected site of neuronal activity. Magn Reson Med 60:548–554, 2008.

Observation of two distinct spatial-temporal BOLD clusters during sensory stimulation in rats.

Neuronal activity evokes changes in local CBF and CBV, whose spatial differences are not fully known. We use the Radial Correlation Contrast (RCC) analysis method with high spatial resolution 100 x 100 x 1000 microm3 data collected with an 11.7 T magnet to differentiate two spatial-temporal BOLD clusters during sensory rat forepaw stimulation and hypothesize that each corresponds to either the CBF or the CBV processes. One cluster, obtained during the time segment of stimulation onset, is characterized by a high positive BOLD signal whereas the other, obtained during the simulation decline time segment, is characterized by a lower positive signal and strong post stimulus undershoot. The average volume of stimulation onset clusters is embedded in the stimulation decline clusters with the latter significantly larger and shifted towards deeper cortical layers. Comparison of amplitude-RCC and cross-correlation analyses performed on equivalent time segments (30 s, 40 images) revealed no differences in cluster size or location, demonstrating that temporal locality is more important than spatial locality in distinguishing between stimulation onset and stimulation decline clusters. We hypothesize that clusters characterized by stimulation onset are highly weighted by local changes in CBF whereas clusters characterized by stimulation decline are more CBV weighted. Moreover, the data suggest that the locations of the highest CBF changes are distinct from the locations of the highest CBV changes. While the former located within stimulation decline clusters and its weight is gradually reduced towards clusters periphery (mainly ventrally), the highest changes in CBV occur in the clusters periphery with only modest changes towards its center.