Layer-specific connectivity revealed by diffusion-weighted functional MRI in the rat thalamocortical pathway

Investigating neural activity from a global brain perspective in-vivo has been in the domain of functional Magnetic Resonance Imaging (fMRI) over the past few decades. The intricate neurovascular couplings that govern fMRI's blood-oxygenation-level-dependent (BOLD) functional contrast are invaluable in mapping active brain regions, but they also entail significant limitations, such as non-specificity of the signal to active foci. Diffusion-weighted functional MRI (dfMRI) with relatively high diffusion-weighting strives to ameliorate this shortcoming as it offers functional contrasts more intimately linked with the underlying activity. Insofar, apart from somewhat smaller activation foci, dfMRI's contrasts have not been convincingly shown to offer significant advantages over BOLD-driven fMRI, and its activation maps relied on significant modelling. Here, we study whether dfMRI could offer a better representation of neural activity in the thalamocortical pathway compared to its (spin-echo (SE)) BOLD counterpart. Using high-end forepaw stimulation experiments in the rat at 9.4 T, and with significant sensitivity enhancements due to the use of cryocoils, we show for the first time that dfMRI signals exhibit layer specificity, and, additionally, display signals in areas devoid of SE-BOLD responses. We find that dfMRI signals in the thalamocortical pathway cohere with each other, namely, dfMRI signals in the ventral posterolateral (VPL) thalamic nucleus cohere specifically with layers IV and V in the somatosensory cortex. These activity patterns are much better correlated (compared with SE-BOLD signals) with literature-based electrophysiological recordings in the cortex as well as thalamus. All these findings suggest that dfMRI signals better represent the underlying neural activity in the pathway. In turn, these advanatages may have significant implications towards a much more specific and accurate mapping of neural activity in the global brain in-vivo.