Faculty Bibliography

Motion during magnetic resonance imaging (MRI) scans routinely results in undesirable image artifact or blurring. Since high-resolution, three-dimensional (3D) imaging of the mouse requires long scan times for satisfactory signal-to-noise ratio (SNR) and image quality, motion-related artifacts are likely over much of the body and limit applications of mouse MRI. In this investigation, we explored the use of self-gated imaging methods and image coregistration for improving image quality in the presence of motion. Self-gated signal results from a modified 3D gradient-echo sequence showed detection of periodic respiratory and cardiac motion in the adult mouse-with excellent comparison to traditional measurements, sensitivity to respiration-induced tissue changes in the brain, and even detection of embryonic cardiac motion in utero. Serial image coregistration with rapidly-acquired, low-SNR volumes further enabled detection and correction of bulk changes in embryo location during in utero imaging sessions and subsequent reconstruction of high-quality images. These methods, in combination, are shown to expand the range of applications for 3D mouse MRI, enabling late-stage embryonic heart imaging and introducing the possibility of longitudinal developmental studies from embryonic stages through adulthood. Magn Reson Med, 2009. (c) 2009 Wiley-Liss, Inc

The availability of an electrocardiogram (ECG) waveform in the adult mouse has permitted the measurement of fast, dynamic cardiac events where data acquisition is synchronized to the R-wave of the ECG waveform. These methods can easily attain one thousand frames/s at ultrasound frequencies greater than 20 MHz. With the heart being the first organ to develop, normal cardiovascular function is crucial to the viability of the developing embryo. Thus, translating such methodologies to analyze embryonic cardiovascular development would add crucial information in mouse models of congenital heart disease which are embryonic lethal. Obtaining an ECG signal from mouse embryos is impractical. Therefore, in this study, preliminary results are presented which derive a cardiac-trigger signal from Doppler blood-flow waveforms. A continuous wave 40 MHz Doppler ultrasound system was used to acquire the Doppler waveforms and a real-time algorithm was developed to process the Doppler waveforms and generate a trigger. Validation studies revealed that a heart rate can be reliably measured and that the Doppler trigger algorithm was robust enough to follow changes in the blood flow. Preliminary data showed that Doppler-derived triggers can be used for highframe-rate prospective imaging of the early embryonic mouse heart

Amyloid plaques are a characteristic feature in Alzheimer's disease (AD). A novel non-toxic contrast agent is presented, Gd-DTPA-K6Abeta1-30, which is homologous to Abeta, and allows plaque detection in vivo. muMRI was performed on AD model mice and controls prior to and following intracarotid injection with Gd-DTPA-K6Abeta1-30 in mannitol solution, to transiently open the blood-brain barrier. A gradient echo T2(*)-weighted sequence was used to provide 100mum isotropic resolution with imaging times of 115min. The scans were examined with voxel-based analysis (VBA) using statistical parametric mapping, for un-biased quantitative comparison of ligand-injected mice and controls. The results indicate that: (1) Gd-DTPA-K6Abeta1-30 is an effective, non-toxic, ligand for plaque detection when combined with VBA (p</=0.01-0.001), comparing pre and post-ligand injection scans. (2) Large plaques can be detected without the use of a contrast agent and this detection co-localizes with iron deposition. (3) Smaller, earlier plaques require contrast ligand for MRI visualization. Our ligand when combined with VBA may be useful for following therapeutic approaches targeting amyloid in transgenic mouse models

The mouse is the preferred model organism for genetic studies of mammalian brain development. MRI has potential for in utero studies of mouse brain development, but has been limited previously by challenges of maximizing image resolution and contrast while minimizing artifacts due to physiological motion. Manganese (Mn)-enhanced MRI (MEMRI) studies have demonstrated central nervous system (CNS) contrast enhancement in mice from the earliest postnatal stages. The purpose of this study was to expand MEMRI to in utero studies of the embryonic CNS in combination with respiratory gating to decrease motion artifacts. We investigated MEMRI-facilitated CNS segmentation and three-dimensional (3D) analysis in wild-type mouse embryos from midgestation, and explored effects of Mn on embryonic survival and image contrast. Motivated by observations that MEMRI provided an effective method for visualization and volumetric analysis of embryonic CNS structures, especially in ventral regions, we used MEMRI to examine Nkx2.1 mutant mice that were previously reported to have ventral forebrain defects. Quantitative MEMRI analysis of Nkx2.1 knockout mice demonstrated volumetric changes in septum (SE) and basal ganglia (BG), as well as alterations in hypothalamic structures. This method may provide an effective means for in utero analysis of CNS phenotypes in a variety of mouse mutants

Manganese-enhanced MRI (MEMRI) has been developed to image brain activity in small animals, including normal and genetically modified mice. Here, we report the use of a MEMRI-based statistical parametric mapping method to analyze sound-evoked activity in the mouse auditory midbrain, the inferior colliculus (IC). Acoustic stimuli with defined frequency and amplitude components were shown to activate and enhance neuronal ensembles in the IC. These IC activity patterns were analyzed quantitatively using voxel-based statistical comparisons between groups of mice with or without sound stimulation. Repetitive 40-kHz pure tone stimulation significantly enhanced ventral IC regions, which was confirmed in the statistical maps showing active regions whose volumes increased in direct proportion to the amplitude of the sound stimuli (65 dB, 77 dB, and 89 dB peak sound pressure level). The peak values of the activity-dependent MEMRI signal enhancement also increased from 7% to 20% for the sound amplitudes employed. These results demonstrate that MEMRI statistical mapping can be used to analyze both the 3D spatial patterns and the magnitude of activity evoked by sound stimuli carrying different energy. This represents a significant advance in the development of MEMRI for quantitative and unbiased analysis of brain function in the deep brain nuclei of mice

Prospective imaging with an annular-array scan was implemented to image the mouse heart at high frame (> 200 frames/s, fps). M-mode data were acquired at a of spatial locations using ECG gating. The pulse-repetition of the M-mode data determined the frame rate of the , reassembled B-mode images. The hearts of adult mice were imaged with a 40-MHz annular array. Annulararray were acquired at an effective frame rate of 500 /s spanning 0.5 s. Data from the annular array were acquired all transmit-to-receive pairs and synthetically focused post-processing. The resulting images showed that multiple cycles could be clearly resolved using prospective imaging that synthetic focusing improved the image resolution and of the right ventricle, interventricular septum, posterior of the left ventricle (LV), and papillary muscles of the LV compared to fixed-focused imaging.

The cortex is thought to be the primary site of sensory plasticity, particularly during development. Here, we report that large-scale reorganization of the mouse auditory midbrain tonotopic map is induced by a specific sound-rearing environment consisting of paired low- (16 kHz) and high-frequency (40 kHz) tones. To determine the potential for plasticity in the mouse auditory midbrain, we used manganese-enhanced MRI to analyze the midbrain tonotopic maps of control mice during normal development and mice reared in the two-tone (16 + 40 kHz) environment. We found that the tonotopic map emerged during the third postnatal week in normal mice. Before 3 weeks, a larger percentage of auditory midbrain responded to each of the suprathreshold test frequencies, despite the fact that the primary afferent projections are in place even before hearing onset. By 3 weeks, the midbrain tonotopic map of control mice was established, and manganese-enhanced MRI showed a clear separation between the 16- and 40-kHz responses. Two-tone rearing dramatically altered the appearance of these discrete frequency-specific responses. A significant volume of the auditory midbrain became responsive to both rearing frequencies, resulting in a large-scale reorganization of the tonotopic map. These results indicate that developmental plasticity occurs on a much greater scale than previously appreciated in the mammalian auditory midbrain

Imaging of the mammalian cardiac right ventricle (RV) is particularly challenging, especially when a two-dimensional method such as conventional histology is used to evaluate the morphology of this asymmetric, crescent-shaped chamber. MRI may improve the characterization of mutants with RV phenotypes by allowing analysis of the samples in any plane and by facilitating three-dimensional image reconstruction. MRI was used to examine the conditional knockout Cx43-PCKO mouse line known to have RV malformations. To help delineate the cardiovascular system and facilitate identification of the right ventricular outflow tract (RVOT), embryonic day (E) 17.5 embryos were perfusion fixed through the umbilical vein followed by a gadolinium-based contrast agent mixed in 7% gelatin. Micro-MRI experiments were performed at 7 T and followed by paraffin embedding of specimens, histological sectioning and hematoxylin and eosin (H&E) staining. Imaging of up to four embryos simultaneously allowed for higher throughput than traditional individual imaging techniques, while intravascular contrast afforded excellent signal-to-noise characteristics. All control embryos (n = 4) and heterozygous Cx43 knockout embryos (n = 4) had normal-appearing right ventricular outflow tract contours by MRI. Obvious abnormalities in the RVOT, including abnormal bulging and infiltration of contrast into the wall of the RV, were seen in three out of four Cx43-PCKO mutants with MRI. Furthermore, three-dimensional reconstruction of MR images with orthogonal projections as well as maximum-intensity projection allowed for visualization of the relationship of infundibular bulging segments to the pulmonary trunk in Cx43-PCKO mutant hearts. The addition of MRI to standard histology in the characterization of RV malformations in mutant mouse embryos aids in the assessment and understanding of morphologic abnormalities. Flexibility in the viewing of MR images, which can be retrospectively sectioned in any desired orientation, is particularly useful in the investigation of the RV, an asymmetric chamber that is difficult to analyze with two-dimensional techniques.

Mice are used in many studies to determine the role of genetic and molecular factors in mammalian development and human congenital diseases. MRI has emerged as a major method for analyzing mutant and transgenic phenotypes in developing mice, at both embryonic and neonatal stages. Progress in this area is reviewed, with emphasis on the use of MRI to analyze cardiovascular and neural development in mice. Comparisons are made with other imaging technologies, including optical and ultrasound imaging, discussing the potential strengths and weaknesses of MRI and identifying the future challenges for MRI in mouse developmental biology.

Few experimental and complementary theoretical studies have investigated high-frequency (>20 MHz) nonlinear responses from polymer-shelled ultrasound contrast agents. Three polymer agents with different shell properties were examined for their single-bubble backscatter when excited with a 40 MHz tone burst. Higher-order harmonic responses were observed for the three agents; however, their occurrence was at least partly due to nonlinear propagation. Only one of the agents (1.1 microm mean diameter) showed a subharmonic response for longer excitations (approximately 10-15 cycles) and midlevel pressure excitations ( 2.5 MPa). Theoretical calculations of the backscattered spectrum revealed behavior similar to the experimental results in specific parameter regimes