Validity and specificity of BOLD effects and their correction in 1H-fMRS

Purpose This study aimed to characterize blood oxygen level-dependent (BOLD) effects in proton magnetic resonance ( 1 H-MR) spectra obtained during optogenetic activation of the rat forelimb cortex to correct and estimate the accurate changes in metabolite concentration. Methods For a more comprehensive understanding of BOLD effects detected with functional magnetic resonance spectroscopy (fMRS) and to optimize the correction method, a 1 Hz line-narrowing effect was simulated. Then, proton functional magnetic resonance spectroscopy ( 1 H-fMRS) data acquired using stimulated echo acquisition mode (STEAM) at 9.4T in rats ( n = 8) upon optogenetic stimulation of the primary somatosensory cortex were utilized. The data were analyzed using MATLAB routines and LCModel. Uncorrected and corrected 1 H-MR spectra from the simulated and in vivo data were quantified and compared. BOLD-corrected difference spectra were also calculated and analyzed. Additionally, the effects of stimulated and non-stimulated water on the quantification of metabolite concentration swere investigated. Results Significant mean increases in water and N-acetylaspartate (NAA) peak heights (+1.1% and +4.5%, respectively) were found to be accompanied by decreased linewidths (−0.5 Hz and −2.8%) upon optogenetic stimulation. These estimates were used for further defining an accurate line-broadening (lb) factor. The usage of a non-data-driven lb introduced false-positive errors in the metabolite concentration change estimates, thereby altering the specificity of the findings. The water and metabolite BOLD contributions were separated using different water scalings within LCModel. Conclusion The linewidth-matching procedure using a precise lb factor remains the most effective approach for accurately quantifying small (±0.3 μmol/g) metabolic changes in 1 H-fMRS studies. A simple and preliminary compartmentation of BOLD effects was proposed, but it will require validation.