Reproducibility of BOLD signal change induced by breath holding

Blood oxygen level dependent (BOLD) contrast is influenced by some physiological factors such as blood flow and blood volume that can be a source of variability in fMRI analysis. Previous studies proposed to use the cerebrovascular response data to normalize or calibrate BOLD maps in order to reduce variability of fMRI data both among brain areas in single subject analysis and across subjects. Breath holding is one of the most widely used methods to investigate the vascular reactivity. However, little is known about the robustness and reproducibility of this procedure. In this study we investigated three different breath holding periods. Subjects were asked to hold their breath for 9, 15 or 21 s in three separate runs and the fMRI protocol was repeated after 15 to 20 days. Our data show that the BOLD response to breath holding after inspiration results in a complex shape due to physiological factors that influence the signal variation with a timing that is highly reproducible. Nevertheless, the reproducibility of the magnitude of the cerebrovascular response to CO2, expressed as amplitude of BOLD signal and number of responding voxels, strongly depends on duration of breath holding periods. Breath holding period of 9 s results in high variability of the magnitude of the response while longer breath holding durations produce more robust and reproducible BOLD responses.