Anesthesia differentially modulates neuronal and vascular contributions to the BOLD signal

Most studies involving BOLD fMRI in basic neuroscience research are conducted with anesthetized animals. This study investigates neural and hemodynamic activity through a combination of experiments comprising BOLD fMRI, optical calcium recordings and ASL in vivo. Patch clamp experiments of neurons were conducted to evaluate electrophysiological correlates of neural activity in vitro. Various anesthetic conditions embracing numerous anesthetic depths evoked by different concentrations of isoflurane (ISO) and different degrees of hypercapnia under a constant stimulus were investigated. We observed that different anesthetic conditions had major impact on the results obtained, particularly that anesthesia could cause a massive divergence of different experimental modalities. In ventilated animals, robust BOLD responses were detectable even with relatively deep anesthesia, while in non-ventilated animals, BOLD responses were not detectable under these conditions. This was most likely due to hypercapnia caused by respiratory depression, as in ventilated animals administered CO2 had the same effect. This observation agreed with measurements of perfusion, which showed that inhaled CO2 increased perfusion significantly, while ISO did not. In optical calcium measurements, higher concentrations of ISO decreased spontaneous neural activity, but not stimulus-evoked responses. This observation was explained by a generally lower excitability of neurons under ISO, which suppressed spontaneous activity, and consequently left more neurons available to fire synchronously in response to a stimulus. Interpreting this phenomenon as an integrated signal of independent single neurons was supported by patch clamp experiments as the number of action potentials (APs) per stimulus was decreased by addition of CO2. Addition of ISO on the other hand had no significant effect. Our results provide an explanation on the cellular level for anesthesia-dependent observations in previous studies of task-induced BOLD and resting state connectivity. They further inform selection of the adequate anesthetic regimen for a given combination of modalities.