47 research outputs found
Altered Neurocircuitry in the Dopamine Transporter Knockout Mouse Brain
The plasma membrane transporters for the monoamine neurotransmitters dopamine, serotonin, and norepinephrine modulate the dynamics of these monoamine neurotransmitters. Thus, activity of these transporters has significant consequences for monoamine activity throughout the brain and for a number of neurological and psychiatric disorders. Gene knockout (KO) mice that reduce or eliminate expression of each of these monoamine transporters have provided a wealth of new information about the function of these proteins at molecular, physiological and behavioral levels. In the present work we use the unique properties of magnetic resonance imaging (MRI) to probe the effects of altered dopaminergic dynamics on meso-scale neuronal circuitry and overall brain morphology, since changes at these levels of organization might help to account for some of the extensive pharmacological and behavioral differences observed in dopamine transporter (DAT) KO mice. Despite the smaller size of these animals, voxel-wise statistical comparison of high resolution structural MR images indicated little morphological change as a consequence of DAT KO. Likewise, proton magnetic resonance spectra recorded in the striatum indicated no significant changes in detectable metabolite concentrations between DAT KO and wild-type (WT) mice. In contrast, alterations in the circuitry from the prefrontal cortex to the mesocortical limbic system, an important brain component intimately tied to function of mesolimbic/mesocortical dopamine reward pathways, were revealed by manganese-enhanced MRI (MEMRI). Analysis of co-registered MEMRI images taken over the 26 hours after introduction of Mn^(2+) into the prefrontal cortex indicated that DAT KO mice have a truncated Mn^(2+) distribution within this circuitry with little accumulation beyond the thalamus or contralateral to the injection site. By contrast, WT littermates exhibit Mn^(2+) transport into more posterior midbrain nuclei and contralateral mesolimbic structures at 26 hr post-injection. Thus, DAT KO mice appear, at this level of anatomic resolution, to have preserved cortico-striatal-thalamic connectivity but diminished robustness of reward-modulating circuitry distal to the thalamus. This is in contradistinction to the state of this circuitry in serotonin transporter KO mice where we observed more robust connectivity in more posterior brain regions using methods identical to those employed here
Reward circuitry is perturbed in the absence of the serotonin transporter
The serotonin transporter (SERT) modulates the entire serotonergic system in the brain and influences both the dopaminergic and norepinephrinergic systems. These three systems are intimately involved in normal physiological functioning of the brain and implicated in numerous pathological conditions. Here we use high-resolution magnetic resonance imaging (MRI) and spectroscopy to elucidate the effects of disruption of the serotonin transporter in an animal model system: the SERT knock-out mouse. Employing manganese-enhanced MRI, we injected Mn^(2+) into the prefrontal cortex and obtained 3D MR images at specific time points in cohorts of SERT and normal mice. Statistical analysis of co-registered datasets demonstrated that active circuitry originating in the prefrontal cortex in the SERT knock-out is dramatically altered, with a bias towards more posterior areas (substantia nigra, ventral tegmental area, and Raphé nuclei) directly involved in the reward circuit. Injection site and tracing were confirmed with traditional track tracers by optical microscopy. In contrast, metabolite levels were essentially normal in the SERT knock-out by in vivo magnetic resonance spectroscopy and little or no anatomical differences between SERT knock-out and normal mice were detected by MRI. These findings point to modulation of the limbic cortical–ventral striatopallidal by disruption of SERT function. Thus, molecular disruptions of SERT that produce behavioral changes also alter the functional anatomy of the reward circuitry in which all the monoamine systems are involved
The Translational Role of Diffusion Tensor Image Analysis in Animal Models of Developmental Pathologies
Diffusion Tensor Magnetic Resonance Imaging (DTI) has proven itself a powerful technique for clinical investigation of the neurobiological targets and mechanisms underlying developmental pathologies. The success of DTI in clinical studies has demonstrated its great potential for understanding translational animal models of clinical disorders, and preclinical animal researchers are beginning to embrace this new technology to study developmental pathologies. In animal models, genetics can be effectively controlled, drugs consistently administered, subject compliance ensured, and image acquisition times dramatically increased to reduce between-subject variability and improve image quality. When pairing these strengths with the many positive attributes of DTI, such as the ability to investigate microstructural brain organization and connectivity, it becomes possible to delve deeper into the study of both normal and abnormal development. The purpose of this review is to provide new preclinical investigators with an introductory source of information about the analysis of data resulting from small animal DTI studies to facilitate the translation of these studies to clinical data. In addition to an in depth review of translational analysis techniques, we present a number of relevant clinical and animal studies using DTI to investigate developmental insults in order to further illustrate techniques and to highlight where small animal DTI could potentially provide a wealth of translational data to inform clinical researchers
Kuvantamistutkimus Unverricht-Lundborgin taudin (EPM1) hiirimallissa
Unverricht-Lundborg type progressive myoclonus epilepsy (EPM1, OMIM 254800) is an autosomal recessive disorder characterized by onset at the age of 6 to 16 years, incapacitating stimulus-sensitive myoclonus and tonic-clonic epileptic seizures. It is caused by mutations in the gene encoding cystatin B (CSTB). However, the disease processes leading to the observed symptoms are currently unclear. Clinical magnetic resonance imaging (MRI) of the brain has shown neurodegenerative changes and computed tomography data have suggested a bone phenotype. This thesis examined the disease processes and the background of the pathological changes in the brain and the bone, utilizing modern imaging methods and image analysis methodology complemented with experimental data in the mouse model (the Cstb -/- mouse) of the disease.
In order to gain a comprehensive picture of the disease progression in the brain, we performed a longitudinal imaging study in the Cstb -/- mouse. Animals were studied from the pre-symptomatic to fully symptomatic disease stages (1-6 mo). For studying atrophic changes, in vivo MRI volumetry was preformed once a month from 1 to 6 months of age. For investigating white matter (WM) changes, ex vivo diffusion tensor imaging (DTI) was performed at 2, 4 and 6 months. The fractional anisotropy (FA) maps derived from DTI data were analysed using track based spatial statistics (TBSS) that provided us with a hypothesis-free analysis of white matter changes. In vivo volumetry showed progressive volume loss in Cstb-/- mice over time, the rate of which was neither spatially nor temporally uniform over the brain. TBSS revealed progressing FA decrease, suggesting severe and widespread WM damage, with most drastic changes in the cerebellum and the thalamus.
Subsequently the congruence of the observed WM changes between the mouse model and EPM1 patients were evaluated. In vivo DTI data from fully symptomatic adult patients and ex vivo data from fully symptomatic (6 mo) Cstb-/- mice were analysed using TBSS with matching protocols. The results revealed extensive changes with a pattern of chronic WM degeneration in EPM1 patients, with similar alterations detected in Cstb-/- mice. Furthermore, previously unknown brain regions were shown to be affected both in patients and in mice. The imaging data were then used to guide tissue level analyses in mice. The microstructural counterpart of the areas with decreased FA in mice was characterized by immunohistochemistry and transmission electron microscopy. Based on the tissue level findings, the extensive changes identified by DTI in both EPM1 patients and in Cstb-/- mice are probably a consequence of widespread WM loss upon axonal degeneration, and likely contribute to the motor disturbances present in the disease.
Finally, we characterized the bone changes underlying the observed skeletal phenotype in EPM1 patients by performing microtomography (µCT), histology, and in vitro cell culture experiments with the Cstb-/- mouse. Analysis of bone microstructure in Cstb-/- mice using μCT revealed structural changes. Moreover, histology confirmed both structural and functional alterations. The basis of these findings was investigated by studying the functionality of bone resorbing osteoclasts differentiated in vitro from bone marrow. In resorption pit formation assays, less and smaller resorption pits were formed by Cstb-/- osteoclasts, indicating decreased resorptive capacity, likely due to a decrease in osteoclast numbers. These data imply that the skeletal changes in Cstb-/- mice and EPM1 patients are a result of CSTB deficiency leading to altered osteoclast function, and the results would indicate that CSTB has a more substantial role as a modulator of bone metabolism than previously thought.
Our results showed high correlations of the atrophy, WM and bone phenotypes between EPM1 patients and Cstb-/- mice and provided information about brain and tissue level changes present in these pathologies. High correlation between the mouse model and the findings in patients provided further affirmation for the use of the mouse model in EPM1 research. Furthermore, the results provided new insight both into the progression of brain pathology and the processes underlying the bone changes present in the disease. Finally, our research introduced new methodologies for research in mouse models of neurodegenerative diseases, and raised the prospects of future research.Etenevä myoklonusepilepsia (Unverricht-Lundborgin tauti, EPM1) on suomalaiseen tautiperimään kuuluva, autosomaalisesti peittyvästi periytyvä sairaus, jonka ensimmäiset oireet havaitaan 6-15 vuoden iässä. Ensimmäiset oireet ovat useimmiten ulkoisten ärsykkeiden laukaisemia lihasnykäyksiä tai epileptisiä kohtauksia. EPM1 aiheutuu virheistä kystatiini B (CSTB) proteiinia koodaavassa geenissä ja vaikka taudin aiheuttava geenivirhe on selvitetty, oireisiin johtavat tautimekanismit ovat edelleen tuntemattomat. Kliininen magneettikuvantaminen on osoittanut potilaiden aivoissa harmaan aineen vauriota ja tilavuudenalenemaa ja tietokonekerroskuvauksessa potilailla on havaittu kallon luiden paksuuntumista. Tässä väitöskirjatyössä pyrittiin kartoittamaan EPM1:n etenemistä ja havaittujen muutosten aikaansaavia kudos- ja solutason muutoksia käyttäen moderneja kuvantamis- ja kuvankäsittelymenetelmiä sekä histologiaa ja soluviljelykokeita taudin kystatiini B puutteisessa (Cstb -/- hiiri) hiirimallissa. Taudin etenemistä aivoissa selvitettiin suorittamalla pitkittäisseurantaa taudin hiirimallissa. Kuvantaminen aloitettiin oireettomissa hiirissä, ja seurantaa jatkettiin kaikkien taudinkuvaan kuuluvien oireiden ilmenemiseen asti (1-6 kk). EPM1:ssä esiintyvän tilavuudenaleneman etenemistä tutkittiin in vivo magneettikuvantamisella, ja valkean aineen muutoksia ex vivo diffuusiotensorikuvamisella. Käyttäen hypoteesivapaata vokselipohjaista analyysimenetelmää (Track based spatial statistic, TBSS) valkean aineen muutoksia seurattiin analysoimalla fraktionaalisen anisotrofian (FA) muutoksia. Havaitsimme Cstb -/- hiirissä etenevän tilavuudenaleneman, jonka määrä vaihteli eri aivoalueiden välillä. Diffuusiotensorikuvantaminen osoitti etenevään aivokudoksen vaurioon viittaavan FA aleneman, joka ilmeni voimakkaimmin talamuksissa ja pikkuaivossa. Seuraavaksi kartoittamme havaittujen valkean aineen vaurioiden yhtenevyyttä hiirimallissa ja potilaissa. Aikuisten EPM1 potilaiden in vivo ja aikuisten Cstb -/-(6kk) hiirten ex vivo diffuusiotensorikuvantamisdata analysoitiin käyttäen identtisiä TBSS- protokollia. Tulokset osoittivat laaja-alaisia, krooniseen rappeumaan viittaavia valkean aineen muutoksia EPM1-potilaissa, ja hiirimallissa havaittiin vastaavat muutokset. Analyysi paljasti sekä potilaissa, että Cstb -/- hiirissä muutoksia aivoalueilla, joiden ei ole aikaisemmin tiedetty EPM1:ssä vaurioituneen. Havaittujen muutosten alkuperää tutkittiin Cstb -/- hiirissä kudostason tarkastelulla soluleikkeistä immunohistokemiallisin värjäyksin, ja elektronimikroskooppisella analyysillä. Tulokset osoittivat että sekä EPM1 potilaissa että hiirimallissa diffuusiokuvantamisella havaitut valkean aineen muutokset ovat seurausta aksonikadosta, ja ovat todennäköisesti yhteydessä taudissa esiintyviin liikehäiriöihin. EPM1-potilailla havaittujen luumuutosten taustoja selvitettiin Cstb -/- hiirissä tietokonekerrokuvauksella (microtomography; uCT), kudosleikevärjäyksillä ja soluviljelykokein. Tietokonekerroskuvaus osoitti luun hienorakenteen muuttuneen Cstb -/- hiirissä, minkä lisäksi kudosleikevärjäyksissä todettiin luussa sekä rakenteellisia että toiminnallisia muutoksia. Näiden muutosten alkuperää kartoitettiin luuta kierrättävien osteoklastien soluviljelykokein. Kun solujen kykyä hajottaa luuta tutkittiin, havaittiin Cstb -/- hiirten osteoklastien muodostavan vähemmän ja pienempiä resorptiivisia kuoppia. Tämä kertoo osteoklastien heikommasta hajotuskyvystä, ja muutos Cstb -/- hiirissä on todennäköisesti seurausta matalammasta solumäärästä viljelmissä. Vaikuttaa siltä, että sekä EPM1 potilaissa, että taudin hiirimallissa havaitut luumuutokset ovat seurasta kystatiini B puutteen aiheuttamista muutoksista osteoklastien toiminnassa. Nämä tulokset viittaavat siihen, että kystatiini B:llä on aikaisempaa suurempi rooli luun metabolian säätelyssä. Tutkimustuloksemme hiirissä vastasivat potilaissa havaittua aivojen tilavuudenalenemaa, valkean aineen vaurioita ja luulöydöksiä, ja pystyimme osoittamaan niihin johtavat kudos- ja solutason muutokset. Havaintomme valottavat EPM1:n huonosti tunnettua etenevää taudinkuvaa, sekä ennestään tunnettujen muutosten taustaa, ja löydösten yhteneväisyys hiirimallissa ja potilaissa rohkaisee EPM1-tutkimuksen jatkamista Cstb -/- hiirimallissa. Väitöskirjatutkimus esitteli myös uusia menetelmiä aivorappeumatautien hiirimallitutkimuksessa sekä avasi uusia tutkimusalueita EPM1:n taudinkuvan selvittämisessä
Diffusion Tensor Imaging Biomarkers of Brain Development and Disease
<p>Understanding the structure of the brain has been a major goal of neuroscience research over the past century, driven in part by the understanding that brain structure closely follows function. Normative brain maps, or atlases, can be used to understand normal brain structure, and to identify structural differences resulting from disease. Recently, diffusion tensor magnetic resonance imaging has emerged as a powerful tool for brain atlasing; however, its utility is hindered by image resolution and signal limitations. These limitations can be overcome by imaging fixed ex-vivo specimens stained with MRI contrast agents, a technique known as diffusion tensor magnetic resonance histology (DT-MRH). DT-MRH represents a unique, quantitative tool for mapping the brain with unprecedented structural detail. This technique has engendered a new generation of 3D, digital brain atlases, capable of representing complex dynamic processes such as neurodevelopment. This dissertation explores the use of DT-MRH for quantitative brain atlasing in an animal model and initial work in the human brain. </p><p>Chapter 1 describes the advantages of the DT-MRH technique, and the motivations for generating a quantitative atlas of rat postnatal neurodevelopment. The second chapter covers optimization of the DT-MRH hardware and pulse sequence design for imaging the developing rat brain. Chapter 3 details the acquisition and curation of rat neurodevelopmental atlas data. Chapter 4 describes the creation and implementation of an ontology-based segmentation scheme for tracking changes in the developing brain. Chapters 5 and 6 pertain to analyses of volumetric changes and diffusion tensor parameter changes throughout rat postnatal neurodevelopment, respectively. Together, the first six chapters demonstrate many of the unique and scientifically valuable features of DT-MRH brain atlases in a popular animal model.</p><p>The final two chapters are concerned with translating the DT-MRH technique for use in human and non-human primate brain atlasing. Chapter 7 explores the validity of assumptions imposed by DT-MRH in the primate brain. Specifically, it analyzes computer models and experimental data to determine the extent to which intravoxel diffusion complexity exists in the rhesus macaque brain, a close model for the human brain. Finally, Chapter 8 presents conclusions and future directions for DT-MRH brain atlasing, and includes initial work in creating DT-MRH atlases of the human brain. In conclusion, this work demonstrates the utility of a DT-MRH brain atlasing with an atlas of rat postnatal neurodevelopment, and lays the foundation for creating a DT-MRH atlas of the human brain.</p>Dissertatio
Examining the development of brain structure in utero with fetal MRI, acquired as part of the Developing Human Connectome Project
The human brain is an incredibly complex organ, and the study of it traverses many scales across space and time. The development of the brain is a protracted process that begins embryonically but continues into adulthood. Although neural circuits have the capacity to adapt and are modulated throughout life, the major structural foundations are laid in utero during the fetal period, through a series of rapid but precisely timed, dynamic processes. These include neuronal proliferation, migration, differentiation, axonal pathfinding, and myelination, to name a few. The fetal origins of disease hypothesis proposed that a variety of non-communicable diseases emerging in childhood and adulthood could be traced back to a series of risk factors effecting neurodevelopment in utero (Barker 1995). Since this publication, many studies have shown that the structural scaffolding of the brain is vulnerable to external environmental influences and the perinatal developmental window is a crucial determinant of neurological health later in life. However, there remain many fundamental gaps in our understanding of it. The study of human brain development is riddled with biophysical, ethical, and technical challenges. The Developing Human Connectome Project (dHCP) was designed to tackle these specific challenges and produce high quality open-access perinatal MRI data, to enable researchers to investigate normal and abnormal neurodevelopment (Edwards et al., 2022). This thesis will focus on investigating the diffusion-weighted and anatomical (T2) imaging data acquired in the fetal period, between the second to third trimester (22 – 37 gestational weeks). The limitations of fetal MR data are ill-defined due to a lack of literature and therefore this thesis aims to explore the data through a series of critical and strategic analysis approaches that are mindful of the biophysical challenges associated with fetal imaging. A variety of analysis approaches are optimised to quantify structural brain development in utero, exploring avenues to relate the changes in MR signal to possible neurobiological correlates. In doing so, the work in this thesis aims to improve mechanistic understanding about how the human brain develops in utero, providing the clinical and medical imaging community with a normative reference point. To this aim, this thesis investigates fetal neurodevelopment with advanced in utero MRI methods, with a particular emphasis on diffusion MRI. Initially, the first chapter outlines a descriptive, average trajectory of diffusion metrics in different white matter fiber bundles across the second to third trimester. This work identified unique polynomial trajectories in diffusion metrics that characterise white matter development (Wilson et al., 2021). Guided by previous literature on the sensitivity of DWI to cellular processes, I formulated a hypothesis about the biophysical correlates of diffusion signal components that might underpin this trend in transitioning microstructure. This hypothesis accounted for the high sensitivity of the diffusion signal to a multitude of simultaneously occurring processes, such as the dissipating radial glial scaffold, commencement of pre-myelination and arborization of dendritic trees. In the next chapter, the methods were adapted to address this hypothesis by introducing another dimension, and charting changes in diffusion properties along developing fiber pathways. With this approach it was possible to identify compartment-specific microstructural maturation, refining the spatial and temporal specificity (Wilson et al., 2023). The results reveal that the dynamic fluctuations in the components of the diffusion signal correlate with observations from previous histological work. Overall, this work allowed me to consolidate my interpretation of the changing diffusion signal from the first chapter. It also serves to improve understanding about how diffusion signal properties are affected by processes in transient compartments of the fetal brain. The third chapter of this thesis addresses the hypothesis that cortical gyrification is influenced by both underlying fiber connectivity and cytoarchitecture. Using the same fetal imaging dataset, I analyse the tissue microstructural change underlying the formation of cortical folds. I investigate correlations between macrostructural surface features (curvature, sulcal depth) and tissue microstructural measures (diffusion tensor metrics, and multi-shell multi-tissue decomposition) in the subplate and cortical plate across gestational age, exploring this relationship both at the population level and within subjects. This study provides empirical evidence to support the hypotheses that microstructural properties in the subplate and cortical plate are altered with the development of sulci. The final chapter explores the data without anatomical priors, using a data-driven method to extract components that represent coordinated structural maturation. This analysis aims to examine if brain regions with coherent patterns of growth over the fetal period converge on neonatal functional networks. I extract spatially independent features from the anatomical imaging data and quantify the spatial overlap with pre-defined neonatal resting state networks. I hypothesised that coherent spatial patterns of anatomical development over the fetal period would map onto the functional networks observed in the neonatal period. Overall, this thesis provides new insight about the developmental contrast over the second to third trimester of human development, and the biophysical correlates affecting T2 and diffusion MR signal. The results highlight the utility of fetal MRI to research critical mechanisms of structural brain maturation in utero, including white matter development and cortical gyrification, bridging scales from neurobiological processes to whole brain macrostructure. their gendered constructions relating to women
Imaging mouse models of neurodegeneration using multi-parametric MRI
Alzheimer’s disease (AD) is a devastating condition characterised by significant cognitive impairment and memory loss. Transgenic mouse models are increasingly being used to further our knowledge of the cause and progression of AD, and identify new targets for therapeutic intervention. These mice permit the study of specific pathological hallmarks of the disease, including intracellular deposits of hyperphosphorylated tau protein and extracellular amyloid plaques. In order to characterise these transgenic mice, robust biomarkers are required to evaluate neurodegenerative changes and facilitate preclinical evaluation of emerging therapeutics. In this work, a platform for in vivo structural imaging of the rTg4510 mouse model of tauopathy was developed and optimised. This was combined with a range of other clinically relevant magnetic resonance imaging (MRI) biomarkers including: arterial spin labelling, diffusion tensor imaging and chemical exchange saturation transfer. These techniques were applied in a single time-point study of aged rTg4510 mice, as well as a longitudinal study to serially assess neurodegeneration in the same cohort of animals. Doxycycline was administered to a subset of rTg4510 mice to suppress the tau transgene; this novel intervention strategy permitted the evaluation of the sensitivity of MRI biomarkers to the accumulation and suppression of tau. Follow-up ex vivo scans were acquired in order to assess the sensitivity of in vivo structural MRI to the current preclinical gold standard. High resolution structural MRI, when used in conjunction with advanced computational analysis, yielded high sensitivity to pathological changes occurring in the rTg4510 mouse. Atrophy was reduced in animals treated with doxycycline. All other MRI biomarkers were able to discriminate between doxycycline-treated and untreated rTg4510 mice as well as wildtype controls, and provided insight into complimentary pathological mechanisms occurring within the disease process. In addition, this imaging protocol was applied to the J20 mouse model of familial AD. This mouse exhibits widespread plaque formation, enabling the study of amyloid-specific pathological changes. Atrophy and deficits in cerebral blood flow were observed; however, the changes occurring in this model were markedly less than those observed in the rTg4510 mouse. This study was expanded to investigate the early-onset AD observed in individuals with Down’s syndrome (DS) by breeding the J20 mouse with the Tc1 mouse model of DS, permitting the relationship between genetics and neurodegeneration to be dissected. This thesis demonstrates the application of in vivo multi-parametric MRI to mouse models of neurodegeneration. All techniques were sensitive to pathological changes occurring in the models, and may serve as important biomarkers in clinical studies of AD. In addition, in vivo multi-parametric MRI permits longitudinal studies of the same animal cohort. This experimental design produces more powerful results, whilst contributing to worldwide efforts to reduce animal usage with respect to the 3Rs principles
Hierarchical and symmetric infant image registration by robust longitudinal-example-guided correspondence detection: Hierarchical and symmetric infant image registration
To investigate anatomical differences across individual subjects, or longitudinal changes in early brain development, it is important to perform accurate image registration. However, due to fast brain development and dynamic tissue appearance changes, it is very difficult to align infant brain images acquired from birth to 1-yr-old
View-aligned hypergraph learning for Alzheimer’s disease diagnosis with incomplete multi-modality data
AbstractEffectively utilizing incomplete multi-modality data for the diagnosis of Alzheimer's disease (AD) and its prodrome (i.e., mild cognitive impairment, MCI) remains an active area of research. Several multi-view learning methods have been recently developed for AD/MCI diagnosis by using incomplete multi-modality data, with each view corresponding to a specific modality or a combination of several modalities. However, existing methods usually ignore the underlying coherence among views, which may lead to sub-optimal learning performance. In this paper, we propose a view-aligned hypergraph learning (VAHL) method to explicitly model the coherence among views. Specifically, we first divide the original data into several views based on the availability of different modalities and then construct a hypergraph in each view space based on sparse representation. A view-aligned hypergraph classification (VAHC) model is then proposed, by using a view-aligned regularizer to capture coherence among views. We further assemble the class probability scores generated from VAHC, via a multi-view label fusion method for making a final classification decision. We evaluate our method on the baseline ADNI-1 database with 807 subjects and three modalities (i.e., MRI, PET, and CSF). Experimental results demonstrate that our method outperforms state-of-the-art methods that use incomplete multi-modality data for AD/MCI diagnosis