The brain versus vein dilemma in BOLD fMRI has spurred research towards more direct correlates of neuronal activation. Diffusion-weighted fMRI (dfMRI) emerged as a potential alternative 17 years ago. However, its signal origins and utility have been greatly debated. In this work, we combine ultra-high-gradients and spiral readout to characterize dfMRI contrast in humans in parameter spaces (TE, b-value, SNR and resolution) that have never been accessible before. Varying TE over a wide range while keeping the b-value fixed allowed us to detect significant hemodynamic contributions to dfMRI contrast at a b-value of 1200 s/mm2.
Five healthy volunteers were scanned on the Siemens Connectom scanner (3 T, 300 mT/m gradients) with visual stimulation (6 runs of alternating checkerboard at 8 Hz, 20 s OFF/20 s ON) at different b-values and TEs using a 32-channel head coil, FOV: 18.2 cm2, Resolution: 1.75 mm2, 13 coronal slices (2 mm thickness) covering the visual cortex, diffusion weighting direction left-right. The following scans were performed. Subject 1: TE = 22.5, 40, 60 and 80 ms, b = 50, 400, 800 and 1200 s/mm2, Subjects 2-4: TE = 22.5, 60 ms, b = 50, 800 and 1200 s/mm2, Subject 5: TE = 22.5 ms, 2 x b = 50 s/mm2, 4 x b = 1200 s/mm2.
fMRI analysis (FSL FEAT23) was performed on all the datasets with default settings. Percentage signal change (%SC) in the activated voxels was estimated and ANOVA was performed to ascertain the influence of b-value and TE. Further, we relaxed the hemodynamic response function (HRF) assumption24 by allowing voxel-wise lag to the start of the HRF25 (from -3 to +3 seconds, in 0.2 seconds steps) and also performed an analysis without a pre-set response function26 (TENT basis function in AFNI). These analyses help to identify potential spatio-temporal differences in the observed activation patterns.
Significant dfMRI responses were detected in the visual cortex across all TE and b-values used in this study. For each b-value, the activation regions became more spatially localised at shorter TE (Fig. 1). For an activation threshold of Z > 2.3, the signal change resulting from visual stimulation increased with TE and b-value (Fig. 2). The %SC increased significantly as a function of b-value (ANOVA, p = 0.0059, Fig. 3A). However, when the %SC were studied as a function of the number of voxels with greatest activation, this trend did not manifest (Fig. 3B).
The optimum lag HRF analysis showed no major differences in responses compared to the fixed delay HRF analysis (Fig. 4, red and blue plots). However, the TENT analysis, showed visibly different, and possibly earlier responses (Fig. 4, green plots). Both these analyses confirmed that the functional responses are weaker at shorter TEs, but increase with b-value. The analysis of the spatial overlap of activation areas is reported as percentage overlap between a fixed number of voxels ranked by their Z-scores (Fig. 5).
The combination of ultra-high gradients and spiral readout enabled dfMRI acquisitions with very short TEs, thereby significantly suppressing SE-BOLD contributions. Nearly three-fold reduction in SE-BOLD contrast was observed at TE = 22.5 ms compared to TE = 60 ms. This reduction was still two fold at b = 1200 s/mm2, which perhaps indicates significant vascular contributions to dfMRI at b = 1200 s/mm2. Since diffusion-based contributions to dfMRI should be TE-independent, the next step is to identify the lower b-value threshold at which this manifests.
We reproduced the key observation that underpins the hypothesis of cellular origins of dfMRI contrast, namely, an increase in %SC with increasing b-value1,2. However, this analysis does not account for the variations in activation area detected at different b-values. When %SC was analysed in equal number of voxels ranked by Z-scores, this trend did not persist.
The TENT analysis revealed slightly earlier responses compared to HRF-based methods, and may help verify the reported temporal specificity improvements with dfMRI2,27. The high spatial overlap of activation regions at shorter TE and higher b-values could indicate improved specificity of dfMRI when compared to the longer TE and lower b-value SE regime.
In conclusion, the combination of ultra-strong gradients and spiral readout helped to open an expanded parameter space (TE and b-value) for dfMRI. This could be invaluable in future investigations of the dfMRI contrast.
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