Differential expression of the GABA transporters GAT-1 and GAT-3 in brains of rats, cats, monkeys and humans

The homeostasis of GABA is critical to normal brain function. Extracellular levels of GABA are regulated mainly by plasmalemmal gamma-aminobutyric acid (GABA) transporters. Whereas the expression of GABA transporters has been extensively studied in rodents, validation of this data in other species, including humans, has been limited. As this information is crucial for our understanding of therapeutic options in human diseases such as epilepsy, we have compared, by immunocytochemistry, the distributions of the GABA transporters GAT-1 and GAT-3 in rats, cats, monkeys and humans. We demonstrate subtle differences between the results reported in the literature and our results, such as the predominance of GAT-1 labelling in neurons rather than astrocytes in the rat cortex. We note that the optimal localisation of GAT-1 in cats, monkeys and humans requires the use of an antibody against the human sequence carboxyl terminal region of GAT-1 rather than against the slightly different rat sequence. We demonstrate that GAT-3 is localised mainly to astrocytes in hindbrain and midbrain regions of rat brains. However, in species such as cats, monkeys and humans, additional strong immunolabelling of oligodendrocytes has also been observed. We suggest that differences in GAT distribution, especially the expression of GAT-3 by oligodendrocytes in humans, must be accommodated in extrapolating rodent models of GABA homeostasis to humans.

Increased glial-derived neurotrophic factor in the small intestine of rats infected with the tapeworm, Hymenolepis diminuta

The neurotrophin, glial-derived neurotrophic factor (GDNF), is essential for the development of the enteric nervous system (ENS) in both the embryo and neonate and may be important for maintenance and plasticity of ENS. The tapeworm, Hymenolepis diminuta, altered the number of cells containing GNDF in the host's jejunum and ileum. Numbers and locations of GDNF-containing cells were determined by applying monoclonal anti-GDNF antibody to intestinal segments collected from infected and uninfected age-matched rats during the initial 34 days post-infection (dpi). Most cells staining positive for GDNF were present in the lamina propria of the jejunum and ileum from both infected and uninfected rats. The co-localization of staining by the antibodies, anti-GDNF and anti-ED2 (a nuclear specific antibody for resident macrophages) indicated that at least 74% of the cells staining for GDNF were macrophages. Mast cells did not stain with the anti-GDNF antibody. The increased number of GDNF+ cells in the infected rat intestine suggests that this neurotrophin may play a role in the neural and mucosal responses to lumenal tapeworm infection.

The type 2 iodothyronine deiodinase is expressed primarily in glial cells in the neonatal rat brain

Thyroid hormone plays an essential role in mammalian brain maturation and function, in large part by regulating the expression of specific neuronal genes. In this tissue, the type 2 deiodinase (D2) appears to be essential for providing adequate levels of the active thyroid hormone 3,5,3′-triiodothyronine (T3) during the developmental period. We have studied the regional and cellular localization of D2 mRNA in the brain of 15-day-old neonatal rats. D2 is expressed in the cerebral cortex, olfactory bulb, hippocampus, caudate, thalamus, hypothalamus, and cerebellum and was absent from the white matter. At the cellular level, D2 is expressed predominantly, if not exclusively, in astrocytes and in the tanycytes lining the third ventricle and present in the median eminence. These results suggest a close metabolic coupling between subsets of glial cells and neurons, whereby thyroxine is taken up from the blood and/or cerebrospinal fluid by astrocytes and tanycytes, is deiodinated to T3, and then is released for utilization by neurons.

Localization of 17beta-hydroxysteroid dehydrogenase and characterization of testosterone in the brain of the male frog.

Several enzymes involved in the formation of steroids of the pregnene and pregnane series have been identified in the brain, but the biosynthesis of testosterone has never been reported in the central nervous system. In the present study, we have investigated the distribution and bioactivity of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) (EC 1.1.1.62; a key enzyme that is required for the formation of testosterone and estradiol) in the brain of the male frog Rana ridibunda. By using an antiserum against human type I placental 17beta-HSD, immunoreactivity was localized in a discrete group of ependymal glial cells bordering the telencephalic ventricles. HPLC analysis of telencephalon and hypothalamus extracts combined with testosterone radioimmunoassay revealed the existence of two peaks coeluting with testosterone and 5alpha-dihydrotestosterone. After HPLC purification, testosterone was identified by gas chromatography/mass spectrometry. Incubation of telencephalon slices with [3H]pregnenolone resulted in the formation of metabolites which coeluted with progesterone, 17alpha-hydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone, and 5alpha-dihydrotestosterone. The newly synthesized steroid comigrating with testosterone was selectively immunodetected by using testosterone antibodies. These data indicate that 17beta-HSD is expressed in a subpopulation of gliocytes in the frog telencephalon and that telencephalic cells are capable of synthesizing various androgens, including dehydroepiandrosterone, androstenedione, testosterone, and 5alpha-dihydrotestosterone.

Localization of vasopressin mRNA and immunoreactivity in pituicytes of pituitary stalk-transected rats after osmotic stimulation.

The presence of [arginine] vasopressin (AVP) mRNA and AVP immunoreactivity in pituicytes of the neural lobe (NL) of intact and pituitary stalk-transected rats, with and without osmotic stimulation, was examined. AVP mRNA was analyzed by Northern blotting, as well as by in situ hybridization in combination with immunocytochemistry using anti-glial fibrillary acidic protein (GFAP) as a marker for pituicytes. In intact rats, a poly(A) tail-truncated 0.62-kb AVP mRNA was detected in the NL and was found to increase 10-fold with 7 days of continuous salt loading. Morphological analysis of the NL of 7-day salt-loaded rats revealed the presence of AVP mRNA in a significant number of GFAP-positive pituicytes in the NL and in areas most probably containing nerve fibers. Eight days after pituitary stalk transection the NL AVP mRNA diminished in animals given water to drink, whereas in those given 2% saline for 18 h followed by 6 h of water, a treatment repeated on 6 successive days beginning 2 days after surgery, the 0.62-kb AVP mRNA was present. The AVP mRNA in the pituitary stalk-transected, salt-loaded rats showed an exclusive cellular distribution in the NL, indicative of localization in pituicytes. Immunoelectron microscopy showed the presence of AVP immunoreactivity in a subpopulation of pituicytes 7 and 10 days after pituitary stalk transection in salt-loaded animals, when almost all AVP fibers had disappeared from the NL. These data show that a subset of pituicytes in the NL is activated to synthesize AVP mRNA and AVP in response to osmotic stimulation.

D-serine, an endogenous synaptic modulator: localization to astrocytes and glutamate-stimulated release.

Using an antibody highly specific for D-serine conjugated to glutaraldehyde, we have localized endogenous D-serine in rat brain. Highest levels of D-serine immunoreactivity occur in the gray matter of the cerebral cortex, hippocampus, anterior olfactory nucleus, olfactory tubercle, and amygdala. Localizations of D-serine immunoreactivity correlate closely with those of D-serine binding to the glycine modulatory site of the N-methyl-D-aspartate (NMDA) receptor as visualized by autoradiography and are inversely correlated to the presence of D-amino acid oxidase. D-Serine is enriched in process-bearing glial cells in neuropil with the morphology of protoplasmic astrocytes. In glial cultures of rat cerebral cortex, D-serine is enriched in type 2 astrocytes. The release of D-serine from these cultures is stimulated by agonists of non-NMDA glutamate receptors, suggesting a mechanism by which astrocyte-derived D-serine could modulate neurotransmission. D-Serine appears to be the endogenous ligand for the glycine site of NMDA receptors.

Computational approach to localization using global energy minimization

Damage localization induced by strain softening can be predicted by the direct minimization of a global energy function. This article concerns the computational strategy for implementing this principle for softening materials such as concrete. Instead of using heuristic global optimization techniques, our strategies are a hybrid of local optimization methods with a path-finding approach to ensure a global optimum. With admissible nodal displacements being independent variables, it is easy to deal with the geometric (mesh) constraint conditions. The direct search optimization methods recover the localized solutions for a range of softening lattice models which are representative of quasi-brittle structures

Automation of an underground mining vehicle using reactive navigation and opportunistic localization

This paper describes the implementation of an autonomous navigation system onto a 30 tonne Load-Haul-Dump truck. The control architecture is based on a robust reactive wall-following behaviour. To make it purposeful we provide driving hints derived from an approximate nodal-map. For most of the time, the vehicle is driven with weak localization (odometry). This need only be improved at intersections where decisions must be made - a technique we refer to as opportunistic localization. The truck has achieved full-speed autonomous operation at an artificial test mine, and subsequently, at a operational underground mine.

Robust outdoor visual localization using a three-dimensional-edge map

Visual localization systems that are practical for autonomous vehicles in outdoor industrial applications must perform reliably in a wide range of conditions. Changing outdoor conditions cause difficulty by drastically altering the information available in the camera images. To confront the problem, we have developed a visual localization system that uses a surveyed three-dimensional (3D)-edge map of permanent structures in the environment. The map has the invariant properties necessary to achieve long-term robust operation. Previous 3D-edge map localization systems usually maintain a single pose hypothesis, making it difficult to initialize without an accurate prior pose estimate and also making them susceptible to misalignment with unmapped edges detected in the camera image. A multihypothesis particle filter is employed here to perform the initialization procedure with significant uncertainty in the vehicle's initial pose. A novel observation function for the particle filter is developed and evaluated against two existing functions. The new function is shown to further improve the abilities of the particle filter to converge given a very coarse estimate of the vehicle's initial pose. An intelligent exposure control algorithm is also developed that improves the quality of the pertinent information in the image. Results gathered over an entire sunny day and also during rainy weather illustrate that the localization system can operate in a wide range of outdoor conditions. The conclusion is that an invariant map, a robust multihypothesis localization algorithm, and an intelligent exposure control algorithm all combine to enable reliable visual localization through challenging outdoor conditions.

A modified particle filter for simultaneous robot localization and landmark tracking in an indoor environment

This paper presents the implementation of a modified particle filter for vision-based simultaneous localization and mapping of an autonomous robot in a structured indoor environment. Through this method, artificial landmarks such as multi-coloured cylinders can be tracked with a camera mounted on the robot, and the position of the robot can be estimated at the same time. Experimental results in simulation and in real environments show that this approach has advantages over the extended Kalman filter with ambiguous data association and various levels of odometric noise.

Biologically inspired visual landmark processing for simultaneous localization and mapping

This paper illustrates a method for finding useful visual landmarks for performing simultaneous localization and mapping (SLAM). The method is based loosely on biological principles, using layers of filtering and pooling to create learned templates that correspond to different views of the environment. Rather than using a set of landmarks and reporting range and bearing to the landmark, this system maps views to poses. The challenge is to produce a system that produces the same view for small changes in robot pose, but provides different views for larger changes in pose. The method has been developed to interface with the RatSLAM system, a biologically inspired method of SLAM. The paper describes the method of learning and recalling visual landmarks in detail, and shows the performance of the visual system in real robot tests.

Wide-angle visual feature matching for outdoor localization

Wide-angle images exhibit significant distortion for which existing scale-space detectors such as the scale-invariant feature transform (SIFT) are inappropriate. The required scale-space images for feature detection are correctly obtained through the convolution of the image, mapped to the sphere, with the spherical Gaussian. A new visual key-point detector, based on this principle, is developed and several computational approaches to the convolution are investigated in both the spatial and frequency domain. In particular, a close approximation is developed that has comparable computation time to conventional SIFT but with improved matching performance. Results are presented for monocular wide-angle outdoor image sequences obtained using fisheye and equiangular catadioptric cameras. We evaluate the overall matching performance (recall versus 1-precision) of these methods compared to conventional SIFT. We also demonstrate the use of the technique for variable frame-rate visual odometry and its application to place recognition.

Reactive navigation and opportunistic localization for autonomous underground mining vehicles

This paper describes an autonomous navigation system for a large underground mining vehicle. The control architecture is based on a robust reactive wall-following behaviour. To make it purposeful we provide driving hints derived from an approximate nodal-map. For most of the time, the vehicle is driven with weak localization (odometry). This need only be improved at intersections where decisions must be made – a technique we refer to as opportunistic localization. The paper briefly reviews absolute and relative navigation strategies, and describes an implementation of a reactive navigation system on a 30 tonne Load-Haul-Dump truck. This truck has achieved full-speed autonomous operation at an artificial test mine, and subsequently, at a operational underground mine.

Experiments with underwater robot localization and tracking

This paper describes a novel experiment in which two very different methods of underwater robot localization are compared. The first method is based on a geometric approach in which a mobile node moves within a field of static nodes, and all nodes are capable of estimating the range to their neighbours acoustically. The second method uses visual odometry, from stereo cameras, by integrating scaled optical flow. The fundamental algorithmic principles of each localization technique is described. We also present experimental results comparing acoustic localization with GPS for surface operation, and a comparison of acoustic and visual methods for underwater operation.