Investigating neural–hemodynamic coupling and the hemodynamic response function in the awake rat

An understanding of the relationship between changes in neural activity and the accompanying hemodynamic response is crucial for accurate interpretation of functional brain imaging data and in particular the blood oxygen level-dependent (BOLD) fMRI signal. Much physiological research investigating this topic uses anesthetized animal preparations, and yet, the effects of anesthesia upon the neural and hemodynamic responses measured in such studies are not well understood. In this study, we electrically stimulated the whisker pad of both awake and urethane anesthetized rats at frequencies of 1–40 Hz. Evoked field potential responses were recorded using electrodes implanted into the contralateral barrel cortex. Changes in hemoglobin oxygenation and concentration were measured using optical imaging spectroscopy, and cerebral blood flow changes were measured using laser Doppler flowmetry. A linear neural–hemodynamic coupling relationship was found in the awake but not the anesthetized animal preparation. Over the range of stimulation conditions studied, hemodynamic response magnitude increased monotonically with summed neural activity in awake, but not in anesthetized, animals. Additionally, the temporal structure of the hemodynamic response function was different in awake compared to anesthetized animals. The responses in each case were well approximated by gamma variates, but these were different in terms of mean latency (approximately 2 s awake; 4 s anesthetized) and width (approximately 0.6 s awake; 2.5 s anesthetized). These findings have important implications for research into the intrinsic signals that underpin BOLD fMRI and for biophysical models of cortical hemodynamics and neural–hemodynamic coupling.