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Spatially localized neuronal cell lineages in the developing mammalian forebrain.

The role of cell lineage in the organization of the cerebral cortex and striatum of the developing rat forebrain was analysed using retroviral-mediated gene transfer to mark the progeny of individual progenitors. Injections around the onset of neurogenesis (embryonic day 14) produced neuronal- and glial-specific clones in the striatum and cortex. The majority of the neuronal clones were restricted to either the deep or superficial layers of the cortex and to either the striatal patch or matrix compartments of the striatum. Moreover, modeling the distributions of the neuronal clones in various ways revealed that grouping the clones into deep vs superficial cortical compartments and patch vs matrix striatal compartments best accounted for the clone distributions. These results suggest that at the onset of neurogenesis there is a heterogeneity of neuronal progenitors within the proliferative ventricular zone.

Animals

Distinct spatial localization of specific mRNAs in cultured sympathetic neurons.

We examined the subcellular distribution of specific mRNAs in cultured sympathetic neurons. Under appropriate conditions, sympathetic neurons extend both axons and dendrites that are distinguishable by light microscopic and immunocytochemical criteria. In situ hybridization revealed a differential localization of mRNA within dendrites. mRNA encoding MAP2 was abundant in cell bodies and distributed nonhomogeneously throughout the dendritic compartment, but was not detected in axons. In contrast, mRNAs encoding GAP-43 and alpha-tubulin were restricted to the cell body and largely excluded from dendrites as well as axons. Detergent extraction revealed that most dendrite-associated mRNA encoding MAP2 was associated with the Triton X-100 insoluble fraction of the cell. The subset of mRNAs present in the dendritic compartment may encode proteins involved in the morphogenesis and remodeling of dendrites.

Animals

Spatial localization of pre-mRNA transcription and processing within the nucleus.

The organization of transcription, processing, and transport of pre-mRNA within the nucleus is a major unsolved problem in cell biology. Several recent studies have helped to define the localization of specific DNAs, RNAs, and proteins within the nucleus and have led to various models for higher level organization of pre-mRNA metabolism.

Animals

Lateral neck imaging for spatial localization of parathyroid tissue.

Two patients with an ectopic parathyroid adenoma are described. In both cases the lesions were clearly demonstrated in an anterior view on T1-201/Tc-99m scintigraphy, but were not identified during the first surgical exploration. Failure to identify the ectopic adenoma at surgery in the second patient led to oblique and lateral views being obtained. These views permitted correct depth localization. We, therefore, believe that lateral and oblique views can be of assistance in improving preoperative localization of an ectopic parathyroid adenoma.

Adenoma

Spatial localization of the K+ channel selectivity filter by mutant cycle-based structure analysis.

The structurally well-characterized scorpion toxin Agitoxin2 inhibits ion permeation through Shaker K+ channels by binding to the external pore entryway. Scanning mutagenesis identified a set of inhibitor residues critical for making energetic contacts with the channel. Using thermodynamic mutant cycle analysis, we have mapped channel residues relative to the known inhibitor structure. This study constrains the position of multiple channel residues within the pore-forming loops; in one stretch, we have been able to map five out of seven contiguous residues to the inhibitor interaction surface, including those involved in ion selectivity. One interaction in particular, that of K27M on the inhibitor with Y445F on the channel, is unique in that it depends on the K+ ion concentration. These results reveal a shallow vestibule formed by the pore loops at the K+ channel entryway. The selectivity filter is located at the center of the vestibule close to (approximately 5 A) the extracellular solution.

Amino Acid Sequence

Ethanol and spatial localization.

Water (Exp. 1) and radial maze (Exp. 2) tasks permitted an evaluation of the relative degree of impairment imposed by ethanol (0, 0.75, 1.5, and 2.0 g/kg) on cognitive mapping vs. cued place learning. The tasks did not require working memory. A strong tendency emerged for ethanol-treated rats to persist in cognitive mapping strategies after the strategies were no longer useful, but there was no indication of a mapping impairment per se. When performance deficits appeared, they were equivalent across mapping and cued place tasks and may have reflected motivational effects of ethanol. In most instances, neither mapping nor cued place tasks were difficult for ethanol-treated animals unless the tasks required abandoning one strategy for another. The tenacity of ethanol-treated rats to use cognitive mapping strategies, particularly rats receiving the highest dose, proved consistent and theoretically decisive. The behavioral invariance of ethanol-treated rats is not caused by a cognitive mapping deficit. Rather, mapping is another domain in which ethanol reduces flexibility.

Animals

Effects of unilateral parietal lesions on spatial localization in the rat.

In the first of two experiments, rats with left or right parietal lesions and controls were tested in place and landmark navigation in the water maize. Right parietal lesions resulted in deficits in both tasks, but especially landmark navigation. Lateralized effects appeared mainly in latency to find the platform. Experiment 2 investigated the role of the corpus callosum. Split-brain rats with unilateral parietal lesions were tested on the same two tasks. Place and landmark deficits were particularly severe, but lateralization was weaker. Callosum section had its own effect, impairing the learning of both tasks. There appear to be additive effects of unilateral cortical lesions and bisection of the hemispheres. The impairment from left lesions equaled the right-lesion deficit because of the interruption of compensatory information from the intact right hemisphere and the effect of callosum section itself.

Animals

Spatial localization under conflict conditions: is there a single explanation?

Visual--auditory (VA) and visual--proprioceptive (VP) localization conflict paradigms were varied to explore the comparability of the conflict situations. In experiment 1 various attempts were made to decrease the dominance of visual information over proprioceptive and auditory target information. Pairing auditory with proprioceptive information against conflicting visual information did not lessen the visual dominance, nor did dimming the visual field. A 'cognitive' manipulation, in which the subject was led to doubt the reliability of the visual information, reduced visual dominance over audition but not visual dominance over proprioception. This difference between the two conflict situations was further explored and corroborated in experiment 2. In experiment 3 no attempt was made to lead the subject to believe that paired discrepant targets were related, and the visual dominance of audition was strong while the visual dominance of proprioception did not occur. The apparent differences between the VA and VP conflict situations are discussed with regard to the feability of generating a unitary explanation of localization conflict results. Several further factors are discussed that must be explored before undertaking such a unitary formulation.

Cognition

Spatially localized 1H NMR spectra of metabolites in the human brain.

Using a surface coil, we have obtained 1H NMR spectra from metabolites in the human brain. Localization was achieved by combining depth pulses with image-selected in vivo spectroscopy magnetic field gradient methods. 1H spectra in which total creatine (3.03 ppm) has a signal/noise ratio of 95:1 were obtained in 4 min from 14 ml of brain. A resonance at 2.02 ppm consisting predominantly of N-acetylaspartate was measured relative to the creatine peak in gray and white matter, and the ratio was lower in the white matter. The spin-spin relaxation times of N-acetylaspartate and creatine were measured in white and gray matter and while creatine relaxation times were the same in both, the N-acetylaspartate relaxation time was longer in white matter. Lactate was detected in the normoxic brain and the average of three measurements was approximately equal to 0.5 mM from comparison with the creatine plus phosphocreatine peak, which was assumed to be 10.5 mM.

Algorithms

Extraocular proprioception is required for spatial localization in man.

We studied the effect of extraocular muscle deafferentation on the performance of manual pointing to foveated visual targets at different craniotopic locations. Five patients who underwent unilateral thermocoagulation of the trigeminal nerve (V) for trigeminal neuralgia were tested before and after surgery. Only the group of patients with postsurgical corneal anaesthesia, attesting to involvement of the ophthalmic branch (which carries the ocular proprioceptive fibres), presented deficits in accuracy of pointing. The position of initial and final pointing was shifted toward the lesioned side predominantly in the ipsilateral hemispace. These data support a model in which balanced ocular proprioceptive inputs are required for accuracy of visually guided movements in egocentric space.

Afferent Pathways

Differential responses of single reticulospinal cells to spatially localized stimulation of the optic tectum in a teleost fish, Salmo trutta.

To determine whether the topographically organized retinal input to the optic tectum is subsequently mapped onto the reticular formation, the responses of antidromically identified reticulospinal cells to tectal surface stimulation were investigated in 45 decerebrated, paralysed trout. The tectum was stimulated through a silver ball surface electrode at 24 different locations, and extracellular recordings were made from the rhombencephalic brainstem with glass microelectrodes filled with a 10% solution of horseradish peroxidase (HRP) in Tris buffer (pH 7.4). After recording, the HRP was, in some cases, iontophoretically expelled from the pipette to identify its location and visualized in histological sections by a modified Hanker-Yates method. Individual reticulospinal neurons discharged 1-4 spikes at short latency in response to stimulation of each of the 24 tectal locations. From one tectal location per cell, this initial response was followed by a late, sustained burst. With a short stimulus train (6 pulses, 55 Hz) the burst could last for over a second with discharge rates of up to 500 Hz. Sometimes this burst could be evoked from neighbouring tectal locations, but only by greatly increasing the stimulus strength. We conclude that the reticular formation receives a highly divergent monosynaptic connection from all locations of the tectum and that the longer latency, sustained burst response is due to a mapped connection between the tectum and the reticular formation. Since the late burst could be preceded by a silent period lasting for approximately 32 ms, we cannot rule out a dependence on interneurons situated between the tectum and the reticulospinal cells.

Animals

Spatial localization after strabismus surgery: evidence for inflow.

Strabismics pointed to targets (without sight of the hand) before and again after surgery that altered the position of the deviating eye in its orbit. Patients having this surgery for the first time were able to use proprioceptively derived information about the surgically altered eye position. In contrast, patients who had similar operations, but on muscles that had been operated on one or more times in the past, were apparently deprived of this information. The important afference may be supplied by the tendon organs.

Afferent Pathways

Identification, characterization, and spatial localization of two flagellin species in Helicobacter pylori flagella.

Flagellar filaments were isolated from Helicobacter pylori by shearing, and flagellar proteins were further purified by a variety of techniques, including CsCl density gradient ultracentrifugation, pH 2.0 acid disassociation-neutral pH reassociation, and differential ultracentrifugation followed by molecular sieving with a Sephacryl S-500 column or Mono Q anion-exchange column, and purified to homogeneity by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transfer to an Immobilon membrane. Two flagellin species of pI 5.2 and with apparent subunit molecular weights (Mrs) of 57,000 and 56,000 were obtained. N-terminal amino acid analysis showed that the two H. pylori flagellin species were related to each other and shared sequence similarity with the N-terminal amino acid sequence of Campylobacter coli, Bacillus, Salmonella, and Caulobacter flagellins. Analysis of the amino acid composition of the predominant 56,000-Mr flagellin species isolated from two strains showed that it was comparable to the flagellins of other species. The minor 57,000-Mr flagellin species contained a higher content of proline. Immunoelectron microscopic studies with polyclonal monospecific H. pylori antiflagellin antiserum and monoclonal antibody (MAb) 72c showed that the two different-Mr flagellin species were located in different regions of the assembled flagellar filament. The minor 57,000-Mr species was located proximal to the hook, and the major 56,000-Mr flagellin composed the remainder of the filament. Western immunoblot analysis with polyclonal rabbit antisera raised against H. pylori or Campylobacter jejuni flagellins and MAb 72c showed that the 56,000-Mr flagellin carried sequences antigenetically cross-reactive with the 57,000-Mr H. pylori flagellin and the flagellins of Campylobacter species. This antigenic cross-reactivity did not extend to the flagellins of other gram-negative bacteria. The 56,000-Mr flagellin also carried H. pylori-specific sequences recognized by two additional MAbs. The epitopes for these MAbs were not surface exposed on the assembled inner flagellar filament of H. pylori but were readily detected by immunodot blot assay of sodium dodecyl sulfate-lysed cells of H. pylori, suggesting that this serological test could be a useful addition to those currently employed in the rapid identification of this important pathogen.

Amino Acid Sequence