Jan G. Bjaalie

Organization of the pontine nuclei

The pontine nuclei provide the cerebellar hemispheres with the majority of their mossy fiber afferents, and receive their main input from the cerebral cortex. Even though the vast majority of pontine neurons send their axons to the cerebellar cortex, and are contacted monosynaptically by (glutamatergic) corticopontine fibers, the information-processing taking place is not well understood. In addition to typical projection neurons, the pontine nuclei contain putative GABA-ergic interneurons and complex synaptic arrangements. The corticopontine projection is characterized by a precise but highly divergent terminal pattern. Large and functionally diverse parts of the cerebral cortex contribute; in the monkey the most notable exception is the almost total lack of projections from large parts of the prefrontal and temporal cortices. Within corticopontine projections from visual and somatosensory areas there is a de-emphasis of central vision and distal parts of the extremities as compared with other connections of these sensory areas. Subcorticopontine projections provide only a few percent of the total input to the pontine nuclei. Certain cell groups, such as the reticular formation, project in a diffuse manner whereas other nuclei, such as the mammillary nucleus, project to restricted pontine regions only, partially converging with functionally related corticopontine connections. The pontocerebellar projection is characterized by a highly convergent pattern, even though there is also marked divergence. Neurons projecting to a single cerebellar folium appear to be confined to a lamella-shaped volume in the pontine nuclei. The organization of the pontine nuclei suggests that they ensure that information from various, functionally diverse, parts of the cerebral cortex and subcortical nuclei are brought together and integrated in the cerebellar cortex.

Quantitative Histological Validation of Diffusion MRI Fiber Orientation Distributions in the Rat Brain

Diffusion MRI (dMRI) is widely used to measure microstructural features of brain white matter, but commonly used dMRI measures have limited capacity to resolve the orientation structure of complex fiber architectures. While several promising new approaches have been proposed, direct quantitative validation of these methods against relevant histological architectures remains missing. In this study, we quantitatively compare neuronal fiber orientation distributions (FODs) derived from ex vivo dMRI data against histological measurements of rat brain myeloarchitecture using manual recordings of individual myelin stained fiber orientations. We show that accurate FOD estimates can be obtained from dMRI data, even in regions with complex architectures of crossing fibers with an intrinsic orientation error of approximately 5–6 degrees in these regions. The reported findings have implications for both clinical and research studies based on dMRI FOD measures, and provide an important biological benchmark for improved FOD reconstruction and fiber tracking methods.

Is lectin-coupled horseradish peroxidase taken up and transported by undamaged as well as by damaged fibers in the central nervous system?

Uptake and transport of horseradish peroxidase-wheat germ agglutinin conjugate (HRP-WGA) in intact and damaged passing fibers were studied by injections of the medulla and pons in 11 cats. Injections with evidence of damage to olivocerebellar fibers and cranial nerve fibers invariably lead to retrograde labeling of neurons in the inferior olive and cranial motor nuclei. With staining around--but apparently no damage of--cranial nerve root fibers, no labeling was found in their motor nuclei. Injections limited to the medullary pyramid with slight fiber damage and limited staining lead to faint retrograde labeling of a small number of cells in the ipsilateral sensorimotor cortex. More extensive staining and fiber damage of the pyramid gave a higher number of labeled cells in the ipsilateral sensorimotor cortex. From these experiments we conclude that HRP-WGA is taken up and transported retrogradely with subsequent significant cell labeling in damaged but not in intact fibers. Anterograde transport of HRP-WGA in fibers passing through the injected area was found to take place only for a very short distance, as judged from cases with injections of either the pons or the medullary pyramid interrupting many corticospinal fibers.

The macroglomerular complex of the antennal lobe in the tobacco budworm moth Heliothis virescens : specified subdivision in four compartments according to information about biologically significant compounds

Abstract The functional organisation of the male specific macroglomerular complex in Heliothis virescens has been studied by tip recordings of sensilla trichodea type 1 combined with cobalt-lysine stainings and by intracellular recordings of antennal lobe projection neurons combined with neurobiotin stainings. The antennal lobe, the macroglomerular complex and the stained axons/dendrites were reconstructed by camera-lucida. Some were further computer reconstructed in three dimensions. The results showed that: 1) The macroglomerular complex consisted of four anatomically separated compartments; 2) A large compartment (the cumulus) at the entrance of the antennal nerve received input from receptor neurons responding to the major pheromone component; 3) Another large compartment, located dorso-medially of the cumulus (the dorso-medial compartment) received input from receptor neurons tuned to the second pheromone component; 4) Two ventrally located compartments received input from two receptor neuron types, co-localized in the same sensillum. Each neuron type responded strongest to one of two interspecifically acting signals, shown to interrupt the pheromone attraction. 5) The function of the dorso-medial compartment was further verified by selective arborizations in this compartment by a projection neuron showing strong response to antennal stimulation with the second pheromone component. At low concentration, the neuron responded synergistically to stimulation with the binary pheromone mixture.

Chapter 13 Salient anatomic features of the cortico-ponto-cerebellar pathway

Recent studies of the primate corticopontine projection show that the neocerebellum--in addition to connections from motor and sensory areas--receives connections from various association areas of the cerebral cortex, some of which are thought to be primarily engaged in cognitive tasks. The quantities of such connections in relation to those from more clearly motor-related parts of the cortex need to be more precisely determined, however. Furthermore, the anatomic data on origin of corticopontine fibers needs to be supplemented with physiological experiments to clarify their functional properties at the single-cell level. For example, nothing is known of the functional role of the large input from the cingulate gyrus, nor is the input from the posterior parietal cortex physiologically characterized. Finally, the scarcity of corticopontine connections from the prefrontal cortex in the monkey (and probably also in man) may not seem readily compatible with a prominent role of the neocerebellum in certain cognitive tasks. We discuss data--in particular from three-dimensional reconstructions--indicating that both corticopontine projects and pontocerebellar neurons are arranged in a lamellar pattern. Corticopontine and pontocerebellar lamellae have similar shapes and orientations but appear to differ in other respects. Corticopontine terminal fields are sharply delimited, apparently without gradual overlap between projections from different sites in the cortex, whereas pontocerebellar lamellae are more fuzzy and exhibit gradual overlap of neuronal populations projecting to different targets. In spite of the sharpness of the corticopontine projection, there may be many opportunities for convergence of inputs from different parts of the cortex. Thus, the wide divergence of corticopontine projections produces many sites of overlap, and extensive interfaces between different terminal fields enabling convergence of inputs onto each neuron. We suggest that the lamellar arrangement of corticopontine terminal fields and of pontocerebellar neurons serve to create diversity of pontocerebellar neuronal properties. Thus, each small part of the cerebellar cortex would receive a specific combination of messages from many different sites in the cerebral cortex. The spatial arrangement of cerebrocerebellar connections have to be understood both in terms of fairly simple large-scale, gradual topographic relationships and an apparently highly complex pattern of divergence and convergence. Developmental studies of corticopontine and of pontocerebellar projections together with three-dimensional reconstructions in adults suggest that the highly complex adult connectional pattern may be created by simple rules operating during development.

In vivo tracing of major rat brain pathways using manganese-enhanced magnetic resonance imaging and three-dimensional digital atlasing.

The magnetic resonance imaging (MRI)-detectable T1 contrast agent manganese (Mn2+) has recently been introduced as a neural tracer in rodents, birds, and monkeys. We have tested to what extent this in vivo method is useful for three-dimensional (3-D) survey of connectivity patterns in the rat somatosensory system. A commonly available 3 T human clinical MRI scanner was used to trace neural pathways following focal injection of manganese chloride (MnCl2) in the somatosensory cortex. Six to 10 h after MnCl2 injection, we found significant signal enhancement in major projection systems, including corticocortical, corticostriatal, corticothalamic, corticotectal, corticopontine, and corticospinal pathways. To facilitate the assignment of anatomic localization to the observed Mn2+ signal enhancement, we registered the MRI data with a 3-D digital reconstruction of a stereotaxic rat brain atlas. Across-animal comparison using the digital model allowed demonstration of a corticothalamic 3-D topographic organization in agreement with previously published two-dimensional topographic schemes based on classical neural tracing data. We conclude that anterograde MnCl2/MRI tracing allows rapid analysis of topographic organization across multiple brain regions. The method allows a higher data throughput for 3-D studies of large-scale brain connectivity than conventional methods based on tissue sectioning.

The concentrations and distributions of three C-terminal variants of the GLT1 (EAAT2; slc1A2) glutamate transporter protein in rat brain tissue suggest differential regulation

The neurotransmitter glutamate is inactivated by cellular uptake; mostly catalyzed by the glutamate transporter GLT1 (slc1a2, excitatory amino acid transporter [EAAT2]) subtype which is expressed at high levels in brain astrocytes and at lower levels in neurons. Three coulombs-terminal variants of GLT1 exist (GLT1a, GLT1b and GLT1c). Their cellular distributions are currently being debated (that of GLT1b in particular). Here we have made antibodies to the variants and produced pure preparations of the individual variant proteins. The immunoreactivities of each variant per amount of protein were compared to that of total GLT1 immunoisolated from Wistar rat brains. At eight weeks of age GLT1a, GLT1b and GLT1c represented, respectively 90%+/-1%, 6+/-1% and 1%+/-0.5% (mean+/-SEM) of total hippocampal GLT1. The levels of all three variants were low at birth and increased towards adulthood, but GLT1a increased relatively more than the other two. At postnatal day 14 the levels of GLT1b and GLT1c relative to total GLT1 were, respectively, 1.7+/-0.1 and 2.5+/-0.1 times higher than at eight weeks. In tissue sections, antibodies to GLT1a gave stronger labeling than antibodies to GLT1b, but the distributions of GLT1a and GLT1b were similar in that both were predominantly expressed in astroglia, cell bodies as well as their finest ramifications. GLT1b was not detected in nerve terminals in normal brain tissue. The findings illustrate the need for quantitative measurements and support the notion that the importance of the variants may not be due to the transporter molecules themselves, but rather that their expression represents the activities of different regulatory pathways.

Waxholm Space atlas of the Sprague Dawley rat brain

Three-dimensional digital brain atlases represent an important new generation of neuroinformatics tools for understanding complex brain anatomy, assigning location to experimental data, and planning of experiments. We have acquired a microscopic resolution isotropic MRI and DTI atlasing template for the Sprague Dawley rat brain with 39 μm isotropic voxels for the MRI volume and 78 μm isotropic voxels for the DTI. Building on this template, we have delineated 76 major anatomical structures in the brain. Delineation criteria are provided for each structure. We have applied a spatial reference system based on internal brain landmarks according to the Waxholm Space standard, previously developed for the mouse brain, and furthermore connected this spatial reference system to the widely used stereotaxic coordinate system by identifying cranial sutures and related stereotaxic landmarks in the template using contrast given by the active staining technique applied to the tissue. With the release of the present atlasing template and anatomical delineations, we provide a new tool for spatial orientation analysis of neuroanatomical location, and planning and guidance of experimental procedures in the rat brain. The use of Waxholm Space and related infrastructures will connect the atlas to interoperable resources and services for multi-level data integration and analysis across reference spaces.

Probing tissue microstructure with restriction spectrum imaging: Histological and theoretical validation.

Water diffusion magnetic resonance imaging (dMRI) is a powerful tool for studying biological tissue microarchitectures in vivo. Recently, there has been increased effort to develop quantitative dMRI methods to probe both length scale and orientation information in diffusion media. Diffusion spectrum imaging (DSI) is one such approach that aims to resolve such information based on the three‐dimensional diffusion propagator at each voxel. However, in practice, only the orientation component of the propagator function is preserved when deriving the orientation distribution function. Here, we demonstrate how a straightforward extension of the linear spherical deconvolution (SD) model can be used to probe tissue orientation structures over a range (or “spectrum”) of length scales with minimal assumptions on the underlying microarchitecture. Using high b‐value Cartesian q‐space data on a rat brain tissue sample, we demonstrate how this “restriction spectrum imaging” (RSI) model allows for separating the volume fraction and orientation distribution of hindered and restricted diffusion, which we argue stems primarily from diffusion in the extraneurite and intraneurite water compartment, respectively. Moreover, we demonstrate how empirical RSI estimates of the neurite orientation distribution and volume fraction capture important additional structure not afforded by traditional DSI or fixed‐scale SD‐like reconstructions, particularly in gray matter. We conclude that incorporating length scale information in geometric models of diffusion offers promise for advancing state‐of‐the‐art dMRI methods beyond white matter into gray matter structures while allowing more detailed quantitative characterization of water compartmentalization and histoarchitecture of healthy and diseased tissue. Hum Brain Mapp, 2013.

Three-dimensional atlas system for mouse and rat brain imaging data

Tomographic neuroimaging techniques allow visualization of functionally and structurally specific signals in the mouse and rat brain. The interpretation of the image data relies on accurate determination of anatomical location, which is frequently obstructed by the lack of structural information in the data sets. Positron emission tomography (PET) generally yields images with low spatial resolution and little structural contrast, and many experimental magnetic resonance imaging (MRI) paradigms give specific signal enhancements but often limited anatomical information. Side-by-side comparison of image data with conventional atlas diagram is hampered by the 2-D format of the atlases, and by the lack of an analytical environment for accumulation of data and integrative analyses. We here present a method for reconstructing 3-D atlases from digital 2-D atlas diagrams, and exemplify 3-D atlas-based analysis of PET and MRI data. The reconstruction procedure is based on two seminal mouse and brain atlases, but is applicable to any stereotaxic atlas. Currently, 30 mouse brain structures and 60 rat brain structures have been reconstructed. To exploit the 3-D atlas models, we have developed a multi-platform atlas tool (available via The Rodent Workbench, http://rbwb.org) which allows combined visualization of experimental image data within the 3-D atlas space together with 3-D viewing and user-defined slicing of selected atlas structures. The tool presented facilitates assignment of location and comparative analysis of signal location in tomographic images with low structural contrast.