Magnetic resonance imaging has previously demonstrated its potential for indirectly mapping myelin density, either by relaxometric detection of myelin water or magnetization transfer. Here, we investigated whether myelin can be detected and possibly quantified directly. We identified the spectrum of myelin in the spinal cord in situ as well as in myelin lipids extracted via a sucrose gradient method, and investigated its spectral properties. Highresolution solution NMR spectroscopy showed the extract composition to be in agreement with myelin's known chemical make-up. The 400-MHz 1 H spectrum of the myelin extract, at 20°C (room temperature) and 37°C, consists of a narrow water resonance superimposed on a broad envelope shifted ∼3.5 ppm upfield, suggestive of long-chain methylene protons. Superimposed on this signal are narrow components resulting from functional groups matching the chemical shifts of the constituents making up myelin lipids. The spectrum could be modeled as a sum of super-Lorentzians with a T 2 * distribution covering a wide range of values (0.008-26 ms). Overall, there was a high degree of similarity between the spectral properties of extracted myelin lipids and those found in neural tissue. The normalized difference spectrum had the hallmarks of membrane proteins, not present in the myelin extract. Using 3D radially ramp-sampled proton MRI, with a combination of adiabatic inversion and echo subtraction, the feasibility of direct myelin imaging in situ is demonstrated. Last, the integrated signal from myelin suspensions is shown, both spectroscopically and by imaging, to scale with concentration, suggesting the potential for quantitative determination of myelin density. myelin in situ | myelin NMR spectrum | super-Lorentzian fitting | ultrashort echo time

Anisotropy of water diffusion in axon tracts, as determined by diffusion-weighted MRI, has been assumed to reflect the restriction of water diffusion across axon membranes. Reduction in this anisotropy has been interpreted as degeneration of axons. These interpretations are based primarily on a priori reasoning that has had little empirical validation. We used the experimental advantages of the sea lamprey spinal cord, which contains several very large axons, to determine whether intraaxonal diffusion is isotropic and whether anisotropy is attributable to restriction of water mobility by axon surface membranes. Through the application of magnetic resonance microimaging, we were able to measure the purely intraaxonal diffusion characteristics of the giant reticulospinal axons (20 -40 m in diameter). The intraaxonal apparent diffusion coefficients of water parallel (longitudinal ADC, l-ADC) and perpendicular (transverse ADC, t-ADC) to the long axis were 0.98 ؎ 0.06 (10 ؊3 mm 2 ͞sec) and 0.97 ؎ 0.11 (10 ؊3 mm 2 ͞sec), respectively. In white matter regions that included multiple axons, l-ADCs were almost identical regardless of axon density in the sampled axon tract. By comparison, t-ADCs were reduced and varied inversely with the number of axons (and thus axolemmas) in a fixed cross-sectional area. Thus, diffusion was found to be isotropic when measured entirely within a single axon and anisotropic when measured in regions that included multiple axons. These findings support the hypothesis that the cell membrane is the primary source of diffusion anisotropy in fiber tracts of the central nervous system. MR microimaging ͉ diffusion-weighted imaging ͉ giant axon ͉ intraaxonal apparent diffusion coefficient ͉ axolemma D iffusion-weighted MRI (DWI) techniques have been widely applied in the white matter (WM) for detection of damage to axons (1-4) and more recently for determining the orientation of fiber tracks (5, 6) in the brain and spinal cord. These applications invoke diffusional anisotropy, a larger apparent diffusion coefficient (ADC) in longitudinal (l-ADC) orientation than in transverse (t-ADC) orientation, in the WM (7).Possible causes of the diffusional anisotropy have been subjects of many studies for over a decade (8-10). To date, although these inferences have been derived from simulation and experimental studies, there have been few studies (11,12) in which the diffusion characteristics of axons have been related directly to axonal anatomy or physiology. This is due largely to limitations in spatial resolution of magnetic resonance (MR) compared with histological methods. In short, it has been possible to obtain MR images of relatively large fields of view, which must be related to the small fields of view achieved with high-power microscopy. Bringing the spatial resolution of MR closer to that of microscopy for diffusion studies will improve correlations of ADCs with histological changes.The nervous system of the sea lamprey (Petromyzon marinus) is unmyelinated. In the spinal cord, the Mauthner and Müll...

Following spinal cord injury, diffusion MRI (DWI) has been shown to detect injury and functionally significant neuroprotection following treatment that otherwise would go undetected with conventional MRI. The underlying histologic correlates to directional apparent diffusion coefficients (ADC) obtained with DWI have not been determined, however, and we address this issue by directly correlating ADC values with corresponding axon morphometry in the normal rat cervical spinal cord. ADC values transverse (perpendicular) and longitudinal (parallel) to axons both correlate with axon counts, however each directional ADC reflects distinct histologic parameters. DWI may therefore be capable of providing specific histologic data regarding the integrity of white matter.

Apparent diffusion coefficients (ADC) were measured in a rat spinal cord weight-drop injury model. After sacrifice, the spinal cords were fixed in situ and excised for MR imaging and ADC measurement. Diffusion is anisotropic in normal gray and white matter. There were significant decreases in ADCs measured along the longitudinal axis of the injured cord and increases in ADCs measured transverse to the cord. Injured segments demonstrated reductions in diffusion anisotropy in the white matter. Diffusion was completely isotropic at the epicenter of the weight-drop injury. Significant decreases in longitudinal ADC and increases in transverse ADC were observed in portions of the cord which appeared normal on conventional spin-echo and calculated T2 images. Thus ADC measurement may complement routine imaging for evaluation of spinal cord injury.