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The development of effective strategies for gene therapy has been hampered by difficulties verifying transgene delivery in vivo and quantifying gene expression non-invasively. Magnetic resonance imaging (MRI) offers high spatial resolution and three-dimensional views, without tissue depth limitations. The iron-storage protein ferritin is a prototype MRI gene reporter. Ferritin forms a paramagnetic ferrihydrite core that can be detected by MRI via its effect on the local magnetic field experienced by water protons. In an effort to better characterize the ferritin reporter for central nervous system applications, we expressed ferritin in the mouse brain in vivo using a neurotropic herpes simplex virus type 1 (HSV-1). We computed three-dimensional maps of MRI transverse relaxation rates in the mouse brain with ascending doses of ferritin-expressing HSV-1. We established that the transverse relaxation rates correlate significantly to the number of inoculated infectious particles. Our results are potentially useful for quantitatively assessing limitations of ferritin reporters for gene therapy applications.

Transplantation of human neural stem cells (hNSCs) is emerging as a viable treatment for stroke related brain injury. However, intraparenchymal grafts do not regenerate lost tissue, but rather integrate into the host parenchyma without significantly affecting the lesion cavity. Providing a structural support for the delivered cells appears important for cell based therapeutic approaches. The non-invasive monitoring of therapeutic methods would provide valuable information regarding therapeutic strategies but remains a challenge. Labeling transplanted cells with metal-based (1)H-magnetic resonance imaging (MRI) contrast agents affects the visualization of the lesion cavity. Herein, we demonstrate that a (19)F-MRI contrast agent can adequately monitor the distribution of transplanted cells, whilst allowing an evaluation of the lesion cavity and the formation of new tissue on (1)H-MRI scans. Twenty percent of cells labeled with the (19)F agent were of host origin, potentially reflecting the re-uptake of label from dead transplanted cells. Both T(2)- and diffusion-weighted MRI scans indicated that transplantation of hNSCs suspended in a gel form of a xenogeneic extracellular matrix (ECM) bioscaffold resulted in uniformly distributed cells throughout the lesion cavity. However, diffusion MRI indicated that the injected materials did not yet establish diffusion barriers (i.e. cellular network, fiber tracts) normally found within striatal tissue. The ECM bioscaffold therefore provides an important support to hNSCs for the creation of de novo tissue and multi-nuclei MRI represents an adept method for the visualization of some aspects of this process. However, significant developments of both the transplantation paradigm, as well as regenerative imaging, are required to successfully create new tissue in the lesion cavity and to monitor this process non-invasively., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Iron oxide-based MRI contrast agents are increasingly being used to noninvasively track cells, target molecular epitopes, and monitor gene expression in vivo. Detecting regions of contrast agent accumulation can be challenging if resulting contrast is subtle relative to endogenous tissue hypointensities. A postprocessing method is presented that yields enhanced positive-contrast images from the phase map associated with T(2)*-weighted MRI data. As examples, the method was applied to an agarose gel phantom doped with superparamagnetic iron-oxide nanoparticles and in vivo and ex vivo mouse brains inoculated with recombinant viruses delivering transgenes that induce overexpression of paramagnetic ferritin. Overall, this approach generates images that exhibit a 1- to 8-fold improvement in contrast-to-noise ratio in regions where paramagnetic agents are present compared to conventional magnitude images. This approach can be used in conjunction with conventional T(2)* pulse sequences, requires no prescans or increased scan time, and can be applied retrospectively to previously acquired data., ((c) 2009 Wiley-Liss, Inc.)

This report presents initial results of a multimodal analysis of tissue volume and microstructure in the brain of an aye-aye (Daubentonia madagascariensis). The left hemisphere of an aye-aye brain was scanned using T2-weighted structural magnetic resonance imaging (MRI) and diffusion-tensor imaging (DTI) prior to histological processing and staining for Nissl substance and myelinated fibers. The objectives of the experiment were to estimate the volume of gross brain regions for comparison with published data on other prosimians and to validate DTI data on fiber anisotropy with histological measurements of fiber spread. Measurements of brain structure volumes in the specimen are consistent with those reported in the literature: the aye-aye has a very large brain for its body size, a reduced volume of visual structures (V1 and LGN), and an increased volume of the olfactory lobe. This trade-off between visual and olfactory reliance is likely a reflection of the nocturnal extractive foraging behavior practiced by Daubentonia. Additionally, frontal cortex volume is large in the aye-aye, a feature that may also be related to its complex foraging behavior and sensorimotor demands. Analysis of DTI data in the anterior cingulum bundle demonstrates a strong correlation between fiber spread as measured from histological sections and fiber spread as measured from DTI. These results represent the first quantitative comparison of DTI data and fiber-stained histology in the brain., ((c) 2005 Wiley-Liss, Inc.)

Sexual dimorphism of brain structures has been reported in some species. We report that sex-dependent developmental structure changes exist in the C57Bl/6(J) mouse, a common model for the genetic analysis of brain function. High resolution, three-dimensional (3D) magnetic resonance microscopy (MRM) images were obtained in intact brains of male and female adult and peripubertal mice. The lateral and third ventricles, hippocampus, amygdala, striatum, and total brain were reconstructed in 3D. As observed in humans, there was overall cerebral growth from peripuberty to adulthood in both sexes. After correcting for the increased brain size, the hippocampus and amygdala were disproportionately larger in adult compared to peripubertal mice. Several sexual dimorphisms were also observed. The lateral ventricles were larger, while the amygdala (the left side in particular) was smaller in females compared to males. Lateral and third ventricles were reduced over time in males only, exhibiting a sex-specific developmental profile. The striatal size was uniform among the groups studied. The surface area of the segmented structures was assayed. Possible shape distortions were detected for the lateral ventricles, hippocampus, and overall brain structure based on a lack of covariance between the surface area and volumetric measurements. Although many sexually dimorphic changes are reported perinatally, our results suggest that there are additional sex-specific transformations that occur around puberty and persist in adulthood.

The feasibility of performing extremely-high resolution somatosensory fMRI in anesthetized mice using BOLD contrast at 11.7 T was investigated. A somatosensory stimulus was applied to the hindlimb of an alpha-chlorolose anesthetized mouse resulting in robust (p < 4 x 10(-3)) BOLD changes in somatosensory cortex and large veins. Percentage modulation of the MR signal in cortex exceeded 7%. Experiments that artificially modulated the inspired oxygen tension were also conducted; the results revealed large, heterogeneous, BOLD contrast changes in the mouse brain. In addition, T(1), T(2), and T(2)* values in gray matter at 11.7 T were evaluated. Discussion of the sensitivity limitations of BOLD fMRI in the tiny mouse central nervous system is presented. These methods show promise for the assessment of neurological function in mouse models of CNS injury and disease., (Copyright 2001 John Wiley & Sons, Ltd.)