Functional MRI in conscious rats using a chronically implanted surface coil.

PURPOSE: To establish procedures for functional MRI (fMRI) in rats without the need for anesthetic agents. MATERIALS AND METHODS: Rats were trained to habituate to restraint in a harness and scanner noise. Under anesthesia, rats were then prepared with a cranial implant that permitted stabilization of the head during subsequent imaging experiments. The cranial implant included an radiofrequency (RF) coil that was used to transmit and receive radiofrequency signals during imaging. Further training was then conducted to habituate the animals to head fixation whilst in the MR scanner. RESULTS: Using this method, we were able to successfully and repeatedly record BOLD fMRI responses to hypercapnia and whisker stimulation in awake rats. Electrical stimulation of the whisker pad produced a ∼7% increase in BOLD signal in the corresponding barrel cortex as well as adjacent negative BOLD responses, whilst hypercapnia produced larger increases in BOLD signal amplitude. CONCLUSION: This methodology leaves the face and limbs free from obstruction, making possible a range of behavioral or sensory stimulation protocols. Further development of this animal model could enable traditional behavioral neuroscience techniques to be combined with modern functional neuroimaging.

Complex spatiotemporal haemodynamic response following sensory stimulation in the awake rat.

Detailed understanding of the haemodynamic changes that underlie non-invasive neuroimaging techniques such as blood oxygen level dependent functional magnetic resonance imaging is essential if we are to continue to extend the use of these methods for understanding brain function and dysfunction. The use of animal and in particular rodent research models has been central to these endeavours as they allow in-vivo experimental techniques that provide measurements of the haemodynamic response function at high temporal and spatial resolution. A limitation of most of this research is the use of anaesthetic agents which may disrupt or mask important features of neurovascular coupling or the haemodynamic response function. In this study we therefore measured spatiotemporal cortical haemodynamic responses to somatosensory stimulation in awake rats using optical imaging spectroscopy. Trained, restrained animals received non-noxious stimulation of the whisker pad via chronically implanted stimulating microwires whilst optical recordings were made from the contralateral somatosensory cortex through a thin cranial window. The responses we measure from un-anaesthetised animals are substantially different from those reported in previous studies which have used anaesthetised animals. These differences include biphasic response regions (initial increases in blood volume and oxygenation followed by subsequent decreases) as well as oscillations in the response time series of awake animals. These haemodynamic response features do not reflect concomitant changes in the underlying neuronal activity and therefore reflect neurovascular or cerebrovascular processes. These hitherto unreported hyperemic response dynamics may have important implications for the use of anaesthetised animal models for research into the haemodynamic response function.

Neurovascular coupling is brain region-dependent.

Despite recent advances in alternative brain imaging technologies, functional magnetic resonance imaging (fMRI) remains the workhorse for both medical diagnosis and primary research. Indeed, the number of research articles that utilise fMRI have continued to rise unabated since its conception in 1991, despite the limitation that recorded signals originate from the cerebral vasculature rather than neural tissue. Consequently, understanding the relationship between brain activity and the resultant changes in metabolism and blood flow (neurovascular coupling) remains a vital area of research. In the past, technical constraints have restricted investigations of neurovascular coupling to cortical sites and have led to the assumption that coupling in non-cortical structures is the same as in the cortex, despite the lack of any evidence. The current study investigated neurovascular coupling in the rat using whole-brain blood oxygenation level-dependent (BOLD) fMRI and multi-channel electrophysiological recordings and measured the response to a sensory stimulus as it proceeded through brainstem, thalamic and cortical processing sites - the so-called whisker-to-barrel pathway. We found marked regional differences in the amplitude of BOLD activation in the pathway and non-linear neurovascular coupling relationships in non-cortical sites. The findings have important implications for studies that use functional brain imaging to investigate sub-cortical function and caution against the use of simple, linear mapping of imaging signals onto neural activity.

Investigating the coupling between stimulation and neural activity: a dynamic modeling approach.

The objective of the present study was to build a dynamic model relating changes in neural responses in rat barrel cortex to an electrical whisker stimulation pulse train of varying frequencies. This work is part of a formal mathematical system currently being developed, which links stimulation to the Blood Oxygen Level Dependent (BOLD) functional Magnetic Resonance Imaging (fMRI) signal. Neural responses were measured in terms of local field potentials, which were then converted into current source density (CSD) data. Responses were found to be strongly suppressed immediately following the first stimulus pulse, before recovering to a steady state, which was maintained throughout the rest of the stimulation. The amplitude of this steady state decreases as the stimulation frequency increases. The model structure is based on the physiological pathway from the rat sensory organ to the cortex. Dynamic linear second order systems are used to model the excitatory as well as the suppressive components of the neural response. The interactions between components contain nonlinear modulations. The model was evaluated against CSD data from experiments with varying stimulation frequency (1-40 Hz), and shows a plausible fit. The model parameters obtained by optimization for different physiological conditions (anaesthetized or awake) were significantly different. Although this is a descriptive model, it may well have some physiological implications.

Simultaneous laser Doppler flowmetry and arterial spin labeling MRI for measurement of functional perfusion changes in the cortex.

This study compares laser Doppler flowmetry (LDF) and arterial spin labeling (ASL) for the measurement of functional changes in cerebral blood flow (CBF). The two methods were applied concurrently in a paradigm of electrical whisker stimulation in the anaesthetised rat. Multi-channel LDF was used, with each channel corresponding to different fiber separation (and thus measurement depth). Continuous ASL was applied using separate imaging and labeling coils at 3 T. Careful experimental set up ensured that both techniques recorded from spatially concordant regions of the barrel cortex, where functional responses were maximal. Strong correlations were demonstrated between CBF changes measured by each LDF channel and ASL in terms of maximum response magnitude and response time-course within a 6-s-long temporal resolution imposed by ASL. Quantitatively, the measurements of the most superficial LDF channels agreed strongly with those of ASL, whereas the deeper LDF channels underestimated consistently the ASL measurement. It was thus confirmed that LDF quantifies CBF changes consistently at a superficial level, and for this case the two methods provided concordant measures of functional CBF changes, despite their essentially different physical principles and spatiotemporal characteristics.

k-space correction of eddy-current-induced distortions in diffusion-weighted echo-planar imaging.

This paper describes a method for correcting eddy-current (EC)-induced distortions in diffusion-weighted echo-planar imaging (DW-EPI). First, reference measurements of EC fields within the EPI acquisition window are performed for DW gradient pulses applied separately along each physical axis of the gradient set and for a range of gradient amplitudes. EC fields caused by the DW gradients of the DW-MRI protocol are then calculated using the reference EC measurements. Finally, these calculated fields are used to correct the respective DW-EPI raw (k-space) data during image reconstruction. The technique was implemented in a small-bore MRI scanner with no digital preemphasis. It corrected EC-induced image distortions in both phantom and in vivo brain diffusion tensor imaging (DTI) data more effectively than commonly used image-based techniques. The method did not increase imaging time, since the same reference EC measurements were used to correct data acquired from different phantoms, subjects, and DTI protocols. Because of the simplicity of the reference EC measurements, the method can easily be implemented in clinical scanners.

How visual stimuli activate dopaminergic neurons at short latency.

Unexpected, biologically salient stimuli elicit a short-latency, phasic response in midbrain dopaminergic (DA) neurons. Although this signal is important for reinforcement learning, the information it conveys to forebrain target structures remains uncertain. One way to decode the phasic DA signal would be to determine the perceptual properties of sensory inputs to DA neurons. After local disinhibition of the superior colliculus in anesthetized rats, DA neurons became visually responsive, whereas disinhibition of the visual cortex was ineffective. As the primary source of visual afferents, the limited processing capacities of the colliculus may constrain the visual information content of phasic DA responses.