Effects of prenatal cocaine exposure on early postnatal rodent brain structure and diffusion properties

Prenatal cocaine exposure has been associated with numerous behavioral phenotypes in clinical populations, including impulsivity, reduced attention, alterations in social behaviors, and delayed language and sensory-motor development. Detecting associated changes in brain structure in these populations has proven difficult, and results have been inconclusive and inconsistent. Due to their more controlled designs, animal models may shed light on the neuroanatomical changes caused by prenatal cocaine; however, to maximize clinical relevance data must be carefully collected using translational methods. The goal of this study was two-fold: 1) determine if prenatal cocaine alters developmental neuroanatomy using methods that are available to human researchers, specifically structural MRI and diffusion tensor imaging; and 2) to determine the feasibility of rodent in vivo neuroimaging for usage in longitudinal studies of developmental disorders. Cocaine-exposed (prenatal days 1–20, 30mg/kg/day) rat pups were sedated and imaged live using diffusion tensor imaging and postmortem (fixed) using magnetic resonance histology on postnatal day 14. Volume and diffusion properties in whole brain as well as specific regions of interest were then assessed from the resulting images. Whole brain analyses revealed that cocaine-exposed animals showed no change in whole brain volume. Additionally, we found alterations in fractional anisotropy across regions associated with reward processing and emotional regulation, especially in the thalamus and globus palladus, as well as sex-dependent effects of cocaine in the right cortex. Reductions in fractional anisotropy were paired with reductions only in axial diffusivity, which preliminarily suggests that the changes observed here may be due to axonal damage, as opposed to reductions in myelination of the affected regions/pathways. Our data indicate that prenatal cocaine may target a number of developing brain structures, but does not result in overt changes to brain volumes. These results highlight not only the brain alterations that result from prenatal cocaine, but also the advancements in live imaging that allow longitudinal study designs in other models

Neuro-imagerie multimodale et multirésolution de cerveaux de souris combinant l’histologie sérielle par tomographie en cohérence optique et l’IRM de diffusion

L’histologie sérielle est une technique d’imagerie permettant d’observer des échantillons entiers à haute résolution. Cette technique consiste à trancher de fines couches de tissu, puis à déplacer l’échantillon sous un objectif de microscope afin d’acquérir autant d’images que nécessaire pour couvrir toute la surface révélée par la coupe. Ce processus est automatisé et est répété jusqu’à ce que tout l’échantillon soit imagé, c’est-à-dire un cerveau de souris dans cette thèse. Couplée à un microscope par tomographie en cohérence optique (OCT), cette modalité est capable de cartographier la distribution spatiale de la matière blanche dans des cerveaux entiers de souris. L’objectif principal de cette thèse était de développer les méthodes de reconstruction nécessaires à l’assemblage en un seul volume des milliers d’images acquises par un système d’histologie massive. De plus, dans cette première phase du projet, des méthodes permettant d’aligner les données sur des images IRM acquises pour les mêmes animaux ont été développées. Cela a permis de mieux comprendre l’origine du contraste optique dans le cerveau et cela offre maintenant la possibilité d’intégrer l’histologie massive dans les études de neuro-imagerie employant des groupes d’animaux. Dans une seconde phase du projet, un microscope à cohérence optique haute résolution a été ajouté au système d’histologie par OCT existant. Cette nouvelle plateforme d’imagerie utilise les images à basse résolution comme repère pour localiser au sein du cerveau les images à haute résolution du second microscope. L’utilité d’une telle plateforme réside dans le fait qu’il est maintenant possible de cibler des régions spécifiques à observer en détail sans avoir à imager un cerveau entier à cette grande résolution, ce qui représenterait plusieurs semaines de mesurage et des quantités immenses de données à assembler. Les données mesurées avec la nouvelle plateforme ont été intégrées à la procédure de reconstruction et d’alignement développé pour la première phase du projet. Ainsi, il a été possible de comparer les images à grande résolution avec les données d’IRM de diffusion acquises pour les mêmes cerveaux de souris. Ceci a permis de confirmer des hypothèses posées lors de l’analyse des données IRM de diffusion à partir de la microscopie. Les méthodes de reconstruction, d’alignement et d’analyse développées, ainsi que la nouvelle plateforme d’histologie sérielle bi-résolution par OCT, offrent enfin la possibilité d’utiliser cette modalité optique pour réaliser des études de groupes animales ou bien pour valider des mesures faites dans le cerveau avec d’autres modalités d’imagerie telle que l’IRM de diffusion.----------ABSTRACT Serial histology is an imaging technique able to observe whole samples at high resolution. This technique involves cutting thin tissue layers, followed by the positioning of the sample under a microscope objective and the acquisition of as many images as necessary to cover the entire area revealed by the cut. This process is automated and is repeated until the entire brain has been imaged. Coupled with an optical coherence tomography (OCT) microscope, this modality is able to map the spatial distribution of white matter in whole mouse brains. The main objective of this thesis was to develop the reconstruction methods necessary for the assembly into a single volume of the thousands of images acquired with a massive histology system. In addition, in this first project phase, methods for aligning data on MRI images acquired for the same animals have been developed. This has led to a better understanding of the optical contrast origin in the brain and it now offers the possibility of integrating massive histology into neuroimaging studies using animal groups. In a second phase of the project, a high resolution optical coherence microscope was added to the existing OCT histology system. This new imaging platform uses low-resolution images as a reference to locate the high-resolution images of the second microscope within the brain. The usefulness of such a platform lies in the fact that it is now possible to target specific regions to observe in detail without having to image an entire brain at this high resolution, which would represent several weeks for measurements and immense quantities of data to assemble. The data measured with the new platform have been incorporated into the reconstruction and alignment procedure developed for the first phase of the project. Thus, it was possible to compare the high resolution images with the diffusion MRI data acquired for the same mouse brains. This made it possible to confirm hypotheses posed during the analysis of diffusion MRI data. The methods of reconstruction, alignment and analysis developed during this thesis, as well as the new dual resolution serial OCT histology platform, finally offer the possibility of using this optical modality to carry out studies of animal groups or to validate measurements made in a brain with other imaging modalities such as diffusion MRI

Automated morphometric analysis and phenotyping of mouse brains from structural µMR images

In light of the utility and increasing ubiquity of mouse models of genetic and neurological disease, I describefully automated pipelines for the investigation of structural microscopic magnetic resonance images of mouse brains – for both high-throughput phenotyping, and monitoring disease. Mouse models offer unparalleled insight into genetic function and brain plasticity, in phenotyping studies; and neurodegenerative disease onset and progression, in therapeutic trials. I developed two cohesive, automatic software tools, for Voxel- and Tensor-Based Morphometry (V/TBM) and the Boundary Shift Integral (BSI), in the mouse brain. V/TBM are advantageous for their ability to highlight morphological differences between groups, without laboriously delineating regions of interest. The BSI is a powerful and sensitive imaging biomarker for the detection of atrophy. The resulting pipelines are described in detail. I show the translation and application of open-source software developed for clinical MRI analysis to mouse brain data: for tissue segmentation into high-quality, subject-specific maps, using contemporary multi-atlas techniques; and for symmetric, inverse-consistent registration. I describe atlases and parameters suitable for the preclinical paradigm, and illustrate and discuss image processing challenges encountered and overcome during development. As proof of principle and to illustrate robustness, I used both pipelines with in and ex vivo mouse brain datasets to identify differences between groups, representing the morphological influence of genes, and subtle, longitudinal changes over time, in particular relation to Down syndrome and Alzheimer’s disease. I also discuss the merits of transitioning preclinical analysis from predominately ex vivo MRI to in vivo, where morphometry is still viable and fewer mice are necessary. This thesis conveys the cross-disciplinary translation of up-to-date image analysis techniques to the preclinical paradigm; the development of novel methods and adaptations to robustly process large cohorts of data; and the sensitive detection of phenotypic differences and neurodegenerative changes in the mouse brai