Longitudinal Magnetic Resonance Imaging Analysis and Histological Characterization after Spinal Cord Injury in Two Mouse Strains with Different Functional Recovery: Gliosis as a Key Factor

International audienceSpinal cord injuries (SCI) are disastrous neuropathologies causing permanent disabilities. The availability of different strains of mice is valuable for studying the pathophysiological mechanisms involved in SCI. However, strain differences have a profound effect on spontaneous functional recovery after SCI. CX3CR1+/eGFP and Aldh1l1-EGFP mice that express green fluorescent protein in microglia/monocytes and astrocytes, respectively, are particularly useful to study glial reactivity. Whereas CX3CR1+/eGFP mice have C57BL/6 background, Aldh1l1-EGFP are in Swiss Webster background. We first assessed spontaneous functional recovery in CX3CR1+/eGFP and Aldh1l1-EGFP mice over 6 weeks after lateral spinal cord hemisection. Second, we carried out a longitudinal follow-up of lesion evolution using in vivo T2-weighted magnetic resonance imaging (MRI). Finally, we performed in-depth analysis of the spinal cord tissue using ex vivo T2-weighted MRI as well as detailed histology. We demonstrate that CX3CR1+/eGFP mice have improved functional recovery and reduced anxiety after SCI compared with Aldh1l1-EGFP mice. We also found a strong correlation between in vivo MRI, ex vivo MRI, and histological analyses of the injured spinal cord in both strain of mice. All three modalities revealed no difference in lesion extension and volume between the two strains of mice. Importantly, histopathological analysis identified decreased gliosis and increased serotonergic axons in CX3CR1+/eGFP compared with Aldh1l1-EGFP mice following SCI. These results thus suggest that the strain-dependent improved functional recovery after SCI may be linked with reduced gliosis and increased serotonergic innervation

A Combination of Ex vivo Diffusion MRI and Multiphoton to Study Microglia/Monocytes Alterations after Spinal Cord Injury

Central nervous system (CNS) injury has been observed to lead to microglia activation and monocytes infiltration at the lesion site. Ex vivo diffusion magnetic resonance imaging (diffusion MRI or DWI) allows detailed examination of CNS tissues, and recent advances in clearing procedures allow detailed imaging of fluorescent-labeled cells at high resolution. No study has yet combined ex vivo diffusion MRI and clearing procedures to establish a possible link between microglia/monocytes response and diffusion coefficient in the context of spinal cord injury (SCI). We carried out ex vivo MRI of the spinal cord at different time-points after spinal cord transection followed by tetrahydrofuran based clearing and examined the density and morphology of microglia/monocytes using two-photon microscopy. Quantitative analysis revealed an early marked increase in microglial/monocytes density that is associated with an increase in the extension of the lesion measured using diffusion MRI. Morphological examination of microglia/monocytes somata at the lesion site revealed a significant increase in their surface area and volume as early as 72 hours post-injury. Time-course analysis showed differential microglial/monocytes response rostral and caudal to the lesion site. Microglia/monocytes showed a decrease in reactivity over time caudal to the lesion site, but an increase was observed rostrally. Direct comparison of microglia/monocytes morphology, obtained through multiphoton, and the longitudinal apparent diffusion coefficient (ADC), measured with diffusion MRI, highlighted that axonal integrity does not correlate with the density of microglia/monocytes or their somata morphology. We emphasize that differential microglial/monocytes reactivity rostral and caudal to the lesion site may thus coincide, at least partially, with reported temporal differences in debris clearance. Our study demonstrates that the combination of ex vivo diffusion MRI and two-photon microscopy may be used to follow structural tissue alteration. Lesion extension coincides with microglia/monocytes density; however, a direct relationship between ADC and microglia/monocytes density and morphology was not observed. We highlighted a differential rostro-caudal microglia/monocytes reactivity that may correspond to a temporal difference in debris clearance and axonal integrity. Thus, potential therapeutic strategies targeting microglia/monocytes after SCI may need to be adjusted not only with the time after injury but also relative to the location to the lesion site

Signal modeling of an MRI ribbon solenoid coil dedicated to spinal cord injury investigations

International audienceNuclear magnetic resonance imaging (NMRI) is a powerful tool for biological investigations. Nevertheless, the imaging resolution performance results in the combination of the magnetic field (B0) and the antenna efficiency. This latter one results in a compromise between the size of the sample, the location of the region of interest and the homogeneity requirement. In the context of spinal cord imaging on mice, a ribbon solenoid coil is used to enhance the efficiency of the MRI experiment. This paper details the calculation of the local magnetization contribution to the induced voltage of MRI coils. The modeling is illustrated on ribbon solenoid antennas used in emitter–receiver mode for the study. The analytical model, which takes into account the emitting mode, the receiving step and the imaging sequence, is compared to the measurement performed on a 9.4 T VARIAN MRI apparatus. The efficiency of the antenna, in terms of signal to noise ratio, is significantly enhanced with respect to a commercial quadrature volumic antenna, given a significant advantage for the study of spinal cord injuries

BOBINES HOMOGÈNES POUR LA RÉSONANCE MAGNÉTIQUE NUCLÉAIRE PORTABLE (RMN) APPLIQUÉE À L'AGROÉCOLOGIE

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Flip-flop method: A new T1-weighted flow-MRI for plants studies

International audienceThe climate warming implies an increase of stress of plants (drought and torrential rainfall). The understanding of plant behavior, in this context, takes a major importance and sap flow measurement in plants remains a key issue for plant understanding. Magnetic Resonance Imaging (MRI) which is well known to be a powerful tool to access water quantity can be used to measure moving water. We describe a novel flow-MRI method which takes advantage of inflow slice sensitivity. The method involves the slice selectivity in the context of multi slice spin echo sequence. Two sequences such as a given slice is consecutively inflow and outflow sensitive are performed, offering the possiblility to perform slow flow sensitive imaging in a quite straigthforward way. The method potential is demonstrated by imaging both a slow flow measurement on a test bench (as low as 10 μm.s−1) and the Poiseuille’s profile of xylemian sap flow velocity in the xylematic tissues of a tomato plant stem

A Combination of Ex vivo Diffusion MRI and Multiphoton to Study Microglia/Monocytes Alterations after Spinal Cord Injury

Central nervous system (CNS) injury has been observed to lead to microglia activation and monocytes infiltration at the lesion site. Ex vivo diffusion magnetic resonance imaging (diffusion MRI or DWI) allows detailed examination of CNS tissues, and recent advances in clearing procedures allow detailed imaging of fluorescent-labeled cells at high resolution. No study has yet combined ex vivo diffusion MRI and clearing procedures to establish a possible link between microglia/monocytes response and diffusion coefficient in the context of spinal cord injury (SCI). We carried out ex vivo MRI of the spinal cord at different time-points after spinal cord transection followed by tetrahydrofuran based clearing and examined the density and morphology of microglia/monocytes using two-photon microscopy. Quantitative analysis revealed an early marked increase in microglial/monocytes density that is associated with an increase in the extension of the lesion measured using diffusion MRI. Morphological examination of microglia/monocytes somata at the lesion site revealed a significant increase in their surface area and volume as early as 72 hours post-injury. Time-course analysis showed differential microglial/monocytes response rostral and caudal to the lesion site. Microglia/monocytes showed a decrease in reactivity over time caudal to the lesion site, but an increase was observed rostrally. Direct comparison of microglia/monocytes morphology, obtained through multiphoton, and the longitudinal apparent diffusion coefficient (ADC), measured with diffusion MRI, highlighted that axonal integrity does not correlate with the density of microglia/monocytes or their somata morphology. We emphasize that differential microglial/monocytes reactivity rostral and caudal to the lesion site may thus coincide, at least partially, with reported temporal differences in debris clearance. Our study demonstrates that the combination of ex vivo diffusion MRI and two-photon microscopy may be used to follow structural tissue alteration. Lesion extension coincides with microglia/monocytes density; however, a direct relationship between ADC and microglia/monocytes density and morphology was not observed. We highlighted a differential rostro-caudal microglia/monocytes reactivity that may correspond to a temporal difference in debris clearance and axonal integrity. Thus, potential therapeutic strategies targeting microglia/monocytes after SCI may need to be adjusted not only with the time after injury but also relative to the location to the lesion site

Flip-flop SEMS sequence.

<p>In presence of a bidirectionnal flow () the second slice of <b>flip</b> will be sensitive to the positive direction inflow () while the second slice of <b>flop</b> will be sensitive to outflow ().</p

Validation setup description.

<p>The Aladdin pumping device pushes the water from the syringe into a silicone tube forming a loop. The silicone tube crossed by opposite directions water flows is inserted within the birdcage coil into the MRI magnet.</p

Flow velocity quantification.

<p>a) A segmentation is applied on a region crossed by an assumed monodirectional flow. b) The formula given by <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0194845#pone.0194845.e043" target="_blank">Eq (16)</a> is applied to the set of <i>T</i>_<i>R</i> experiment to get the best <i>vmax</i> fitting value (here, 0.178<i>mm</i>/<i>s</i> while the expected value would be 0.2<i>mm</i>/<i>s</i>). c) The velocity scale can be added to the flow image.</p

Flip-flop flow-MRI.

<p>Right figure: at 10<i>μm</i>/<i>s</i> and left figure: 50<i>μm</i>/<i>s</i>. The following parameters were used: <i>T</i>_<i>R</i> = 1.2<i>s</i>, <i>T</i>_<i>E</i> = 15<i>ms</i>, <i>FOV</i>_<i>READ</i> = 40<i>mm</i>, <i>FOV</i>_<i>PHASE</i> = 40<i>mm</i>, <i>N</i>_<i>READ</i> = <i>N</i>_<i>PHASE</i> = 256, <i>T</i>_<i>hk</i> = 0.5<i>mm</i>, 2 slices, <i>AVG</i> = 2 and <i>T</i>_<i>acq</i> = 2 × 10<i>min</i>14.</p