Investigating exchange, structural disorder, and restriction in gray matter via water and metabolites diffusivity and kurtosis time-dependence

Abstract Water diffusion-weighted MRI is a very powerful tool for probing tissue microstructure, but disentangling the contribution of compartment-specific structural disorder from cellular restriction and inter-compartment exchange remains an open challenge. In this work, we use diffusion-weighted MR spectroscopy (dMRS) of water and metabolite as a function of diffusion time in vivo in mouse gray matter to shed light on: i) which of these concomitant mechanisms (structural disorder, restriction, and exchange) dominates the MR measurements and ii) with which specific signature. We report the diffusion time-dependence of water with excellent SNR conditions as provided by dMRS, up to a very long diffusion time (500 ms). Water kurtosis decreases with increasing diffusion time, showing the concomitant influence of both structural disorder and exchange. However, despite the excellent experimental conditions, we were not able to clearly identify the nature of the structural disorder (i.e., 1D versus 2D/3D short-range disorder). Measurements of purely intracellular metabolites diffusion time-dependence (up to 500 ms) show opposite behavior to water, with metabolites kurtosis increasing as a function of diffusion time. We show that this is a signature of diffusion restricted in the intracellular space, from which cellular microstructural features such as soma’s and cell projections’ size can be estimated. Finally, by comparing water and metabolite diffusion time-dependencies, we attempt to disentangle the effect of intra/extracellular exchange and structural disorder of the extracellular space (both impacting water diffusion only). Our results suggest a relatively short intra/extracellular exchange time (~1-50 ms) and short-range disorder (still unclear if 1D or 2D/3D) most likely coming from the extracellular compartment. This work provides novel insights to help interpret water diffusion-time dependent measurements in terms of the underlying microstructure of gray matter and suggests that diffusion-time dependent measurements of intracellular metabolites may offer a new way to quantify microstructural restrictions in gray matter.