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Spatial pattern of catalase (Cat2) gene activation in scutella during postgerminative development in maize.

The scutellum of maize is a fully differentiated, nondividing, diploid embryonic tissue. Two distinct structural genes (Cat1 and Cat2) encoding the enzyme catalase (CAT) are differentially expressed in this tissue during postgerminative development. As development proceeds, the expression of Cat1 diminishes, while that of Cat2 is enhanced, leading to the disappearance of the CAT-1 protein and the gradual accumulation of the CAT-2 protein. The present investigation was undertaken to determine whether all scutellar cells may be genetically programmed to activate expression of Cat2 synchronously or whether there is an asynchronous spatial gradient of Cat2 activation. By using immunofluorescence microscopy and anti-CAT-2 IgG, we have found that a gradient of Cat2 activation occurs within the scutellar cell mass during postgerminative development. The gradient of Cat2 activation occurs from the outer perimeter of the tissue inward toward the embryonic axis. To determine a potential site of origin for any putative "triggering signal" for Cat2 activation, we demonstrated that Cat2 is expressed in the single layer of aleurone cells prior to its expression in any other tissue during kernel development. To our knowledge, this is the first observation of a gradient-type spatial pattern of a eukaryote gene activation occurring in a stable, virtually nondividing tissue such as the maize scutellum. The significance of these results with respect to developmental gene regulation is discussed.

Catalase

Establishment and maintenance of stable spatial patterns in lacZ fusion transformants of Polysphondylium pallidum.

Polysphondylium pallidum cells were transformed with a construct containing the Dictyostelium discoideum ecmA promoter fused to a lacZ reporter gene. Two stably transformed lines, one in which beta-galactosidase (beta-gal) is expressed in apical cells of the fruiting body (p63/2.1), and one in which it is expressed in basal cells (p63/D), have enabled us to infer how cells move during aggregation and culmination. Several types of cell movement proposed to occur during slime mold culmination, such as random cell mixing and global cell circulation, can be ruled out on the basis of our observations. Cells of the two transformant lines express beta-gal very early in development. In both cases, stained cells are randomly scattered in a starving population. By mid to late aggregation, characteristic spatial patterns emerge. Marked cells of p63/2.1 are found predominantly at tips of tight aggregates; those of p63/D accumulate at the periphery. These patterns are conserved throughout culmination, showing that marked cells maintain their relative positions within the multicellular mass following aggregation. Neither the apical nor the basal pattern appears to be regulated within the primary sorogen by de novo gene expression or by cell sorting as whorls are formed. However, marked cells within a whorl re-establish the original pattern in secondary sorogens. This must be achieved by cell migration, since beta-gal is not re-expressed.

Cell Movement

Neuronal competition determines the spatial pattern of neuropeptide expression by identified neurons of the leech.

Staining adult and embryonic leech ventral nerve cords with antibodies raised against the molluscan neuropeptides small cardioactive peptide B (SCP) and FMRFamide results in segment-specific and bilaterally asymmetric patterns of cell staining. One immunoreactive neuron, the RAS interneuron, is present in only four rostral segmental ganglia, while another, the CAS interneuron, is restricted to the four most caudal abdominal ganglia and tail. In addition to their segment-specific distributions, only one RAS or CAS cell is found in each segmental ganglion, and they alternate sides between adjacent ganglia (either L-R-L-R or R-L-R-L) with a fidelity of about 95%. This paper utilizes cell deletion techniques to investigate the determination of the asymmetric and alternating pattern of RAS and CAS neurons. We show that developmentally equivalent RAS and CAS homologs are present on both sides of the appropriate ganglia, and that within each ganglion one of the initially paired homologs loses the ability to assume the immunoreactive RAS or CAS fate 2-3 days after axonogenesis has begun. These experiments suggest that there is a competitive interaction between bilateral homologs which ensures that only one mature RAS/CAS neuron is formed per ganglion, and that contralateral RAS/CAS neurons are not required in the same or adjacent ganglia for the determination of the RAS or CAS developmental pathways. Nerve cord transections between ganglia in the CAS domain can alter the spatial pattern of CAS neuron determination, confirming that both bilateral homologs retain the ability to express neuropeptide until late embryonic stages, and suggesting that the alternating pattern of RAS/CAS cells requires communication between adjacent ganglia through the longitudinal connectives.

Animals

Temporal and spatial patterns of proto-oncogene expression at early stages of toxic liver injury in the rat.

Sequential and transient expression of c-fos, c-jun, c-myc, c-Ha-ras and c-Ki-ras proto-oncogene RNA transcripts with zonal heterogeneity was demonstrated in virtually all hepatocytes of adult rat liver by in situ hybridization with single-stranded, [35S]-labeled cRNA probes at various time points after intraperitoneal administration of a single dose of carbon tetrachloride (CCl4). After a brief interval, elevated RNA levels of these genes were also observed in nonparenchymal cells. A second phase of proto-oncogene expression was characterized by high RNA levels in only a fraction of parenchymal cells with preference of mediolobular areas. Distribution and number of these cells were comparable tl hepatocytes expressing the proliferation-associated nuclear antigen Ki-67 72 hours after toxic injury. Oncogene expression in the nonparenchymal compartment correlated with distinct morphologic changes preceding type I procollagen gene expression by desmin-positive perisinusoidal cells, accumulating together with numerous c-fms expressing cells in the areas of hepatocellular necrosis. We conclude that zonal hepatic destruction by carbon tetrachloride induces proto-oncogene expression with distinct temporal and spatial patterns initiated by the most severely damaged hepatocytes. Proto-oncogene products thus represent valuable markers of cellular activation preceding and accompanying various aspects of tissue repair reactions.

Animals

Abnormal spatial patterns of elbow muscle activation in hemiparetic human subjects.

The patterns of electromyographic (EMG) activity in spastic-paretic and contralateral elbow muscles of 10 hemiparetic human subjects were compared during a sequence of graded voluntary isometric contractions against 4 different-sized loads. These loads were orientated successively at 8 different angles over a 360 degree range, referenced to a plane at the wrist orthogonal to the long axis of the forearm. Comparisons of EMG activity recorded from normal and paretic limbs revealed that there were marked differences in the torque angles which evoked significant EMG activity, in the angular range of EMG, in the angle of peak EMG, and in the scaling of EMG magnitude with increasing isometric loads. In severely impaired limbs, there was a marked shift in both the peak EMG angle and the angular domain of EMG activity for both biceps and triceps muscle groups, away from the normal elbow flexion-extension axis towards external humeral rotation and shoulder girdle elevation. The extent of the disturbance in the spatial patterns of EMG activity was closely correlated with the clinical severity of the spastic-paretic disability, which was quantified using a functional scale patterned after that described by Fugl Meyer et al. (1975). The observed patterns of EMG activity in paretic flexor muscles do not conform with established synergistic patterns, such as might be released by excitation of the flexor reflex in a normal limb. Possible origins for the anomalous EMG patterns are discussed.

Adult

Temporal and spatial pattern of differences in microtubule behaviour during Drosophila embryogenesis revealed by distribution of a tubulin isoform.

Immunofluorescence staining of Drosophila embryos with a monoclonal antibody specific for acetylated alpha-tubulin has revealed that acetylated and nonacetylated alpha-tubulin isoforms have different patterns of distribution during early development. Acetylated alpha-tubulin was not detected in either interphase or mitotic spindle microtubules during the rapid early cleavage or syncytial blastoderm divisions. Acetylated alpha-tubulin was first observed as interphase lengthened at the end of syncytial blastoderm, and at cycle 14 was localized to a ring of structures clustered around the interphase nuclei. These structures probably represent a set of stable microtubules involved in nuclear elongation. Absence of detectable acetylated alpha-tubulin prior to cellular blastoderm seems to be due to rapid turnover of microtubule arrays rather than to lack of the enzyme required for modification, since acetylated alpha-tubulin appeared in early embryos when micro-tubules were stabilized by taxol treatment or anoxia. Because acetylated alpha-tubulin seems to be characteristic of stable microtubule arrays, the appearance of the antigen at cycle 14 represents a fundamental change in microtubule behaviour in the somatic cells of the embryo. Acetylated alpha-tubulin was not detected in pole cells during the blastoderm or early gastrula stages, indicating that acetylation of alpha-tubulin is not merely a consequence of cellularization. After the onset of gastrulation, interphase microtubule arrays in most cell types contain acetylated alpha-tubulin. However, cells in mitosis lack antibody staining. The resulting unstained patches reveal the stereotyped spatial pattern of cell division during gastrulation. Although the cells that give rise to the amnioserosa have acetylated alpha-tubulin in their interphase arrays at early gastrulation, by germ band elongation these large, plastic cells completely lack staining with anti-acetylated alpha-tubulin. In contrast, differentiated cell types such as neurones, which have arrays of stable axonal microtubules, stain brightly with the specific antibody. Although acetylated and nonacetylated alpha-tubulin are present in roughly equal amounts by the late stages of embryogenesis, acetylated alpha-tubulin is partitioned into the pellet during centrifugation of extracts of embryos homogenized at 4 degrees C.

Animals

Serial processing of visual spatial patterns in a search paradigm.

Previous experiments in this laboratory employing a search paradigm have found highly significant differences in the detectability of a briefly exposed target pattern as a function of the spatial location of the target when it is presented simultaneously with a number of discriminably different nontarget patterns. These detectability differences, at loci equidistant from the fovea, could not be accounted for by any known variation in retinal spatial resolution or by differential lateral masking effects of the target by nearby nontarget patterns. These observations led to the hypothesis that the target in these experiments was detected by a serial mechanism which "scanned" a persisting but rapidly degrading neural representation of the visual scene with increasing detection failures the later in time the scan processed the location occupied by the target. If this hypothesis is correct, then target detectability should vary inversely with the number of stimuli which must be examined. The present experiment confirmed this expectation. A mathematical model of such a serial scanning process also predicts other, less obvious, effects on target detectability which were observed when the number of nontarget patterns was changed.

Adult

Specificity of spatial patterns of glomerular activation in the mouse olfactory bulb: computer-assisted image analysis of 2-deoxyglucose autoradiograms.

We have developed a computer-assisted method for analyzing the 2-deoxyglucose (2-DG) autoradiograms of mice olfactory bulbs. The purpose of the study was to numerize the maps of glomerular activation in order to achieve a statistical comparison of the glomerular patterns evoked by different stimuli. The spatial distribution of glomerular activation was displayed on unfolded representations of the glomerular layer which were built up using glomerular optical densities (OD) measured systematically within 13 sections per bulb. Each bulbar sample was converted into an 'OD profile'. A matrix composed of 18 OD profiles was submitted to a principal component analysis. The first factor which accounted for 28% of the variance separated unambiguously two clusters corresponding to the bulbs issued from animals stimulated with amylacetate and isovaleric acid, respectively. The second and third factors which accounted for 14% and 12% of the variance segregated the control group (animals exposed to pure air) from the odor-stimulated ones. It was demonstrated that the cluster separation was actually due to the specific spatial distribution of the most-labelled glomeruli. A particular attention was paid to the well-delineated glomerular activation evoked by isovaleric acid. The results demonstrate the specificity and reliability of the glomerular 2-DG patterns. The method should be useful for further comparisons of patterns elicited by larger sets of odorant compounds.

Animals

Do focal neocortical lesions hamper short-term recognition of visual spatial patterns?

A hundred and forty-three patients with focal - mainly neocortical - hemiphere-damage (subdivided in 4 groups according to the side of the damaged hemisphere and the presence/absence of visual field defect) and 70 controls, were given a 20-sec delay recognition test of complex spatial visual patterns. There was one free delay condition and two conditions with interference from visual material of either similar or quite dissimilar patterns to that of memoranda. The results provided by two co-variance analyses are: (i) the comparison of the hemisphere-damaged groups with controls show a significant memory decay of both right groups only when there was perceptual interference from similar patterns; (ii) the comparisons between hemisphere-damaged groups fail to elicit any significant difference in the three memory conditions. There appears to be no specific neocortical area involved in the first steps of memory processing of spatial informations and the interference engendered by interpolated perceptual activity is material-specific.

Adult

Spatial patterns of olfactory bulb single-unit responses to learned olfactory cues in young rats.

1. Neonatal rat pups were classically conditioned to an odor stimulus from postnatal day 1 (PN1) to PN18. Tactile stimulation (stroking) was used as the unconditioned stimulus. On PN19, mitral/tufted cell single-unit responses to the conditioned odor were examined in both conditioned and control pups. Recordings were made from mitral/tufted cells in two regions of the olfactory bulb: 1) an area typically associated with focal [14C]2-deoxyglucose (2-DG) uptake in response to the conditioned odor and 2) an area distant from focal 2-DG uptake to the conditioned odor. Animals were anesthetized with urethane and were naturally respiring during the single-unit recording procedure. 2. Changes in mitral/tufted cell firing rate in response to odors in both bulbar regions and all training groups were classified as either excitatory, suppressive, or no response. This response classification was used to compare response patterns to the conditioned odor between bulbar regions and training groups. 3. Classical conditioning selectively modified the response patterns of mitral/tufted cells to the conditioned odor when those cells were associated with regions of focal 2-DG uptake for that odor. Mitral/tufted cells demonstrated significantly more suppressive and fewer excitatory responses to the conditioned odor than cells in control pups. Response patterns to a novel odor were not similarly modified. 4. Response patterns of mitral/tufted cells distant from the focal region of 2-DG uptake to the conditioned odor were not modified by conditioning compared with control pups. 5. The difference in response pattern between cells in the 2-DG focus and cells distant to the 2-DG focus was apparent within 500 ms of the stimulus onset. Given the respiratory rate of these pups (2 Hz), these data suggest that the modified response pattern occurred on the first inhalation of the learned odor. 6. These data demonstrate that both spatial and temporal patterns of olfactory bulb output neuron activity are used in the coding of olfactory information in the bulb. Furthermore, these spatial/temporal response patterns can be modified by early learning.

Animals

"Chess-board pattern" spatial modulation of magnetization. Assessment of myocardial function.

Heart motion is a complex combination of translation, rotation, and concentric contraction. Evaluation of these complex motions has been difficult using conventional slice-selective methods. Noninvasive tagging of the heart has been obtained by the use of slice-selective radiofrequency pulses. Through spatial modulation of the magnetization the entire image can be labeled in different patterns. Two new pulse sequences are presented, giving a chess-board like spatial modulation. These pulse sequences have several advantages compared with the previously published methods, as the modulation time is half that required to obtain a 2-dimensional grid, the area in the image with high signal intensity was significantly larger, and the radiofrequency power deposition was substantially decreased. By labeling the heart at diastole the chess-board pattern tagging of the heart wall could be followed through systole. Using this method the complex motions of the heart can be mapped.

Computer Simulation

Expression of two members of the Wnt family during mouse development--restricted temporal and spatial patterns in the developing neural tube.

The Wnt gene family encodes a group of cysteine-rich proteins implicated in intercellular signaling during several stages of vertebrate development. This family includes Wnt-1 and Wnt-3, both discovered as activated oncogenes in mouse mammary tumors. Here we describe the molecular cloning of an additional member of the Wnt family, called Wnt-3A, and the spatial and temporal expression pattern of this gene as well as that of its close relative Wnt-3. The putative amino acid sequences of both proteins are almost 90% identical, but in situ hybridization to mouse embryo sections showed highly restricted patterns of expression of Wnt-3 and Wnt-3A, largely in separate areas in the developing nervous system. In the spinal cord Wnt-3 was expressed at low levels in the alar laminae and in the ventral horns, whereas Wnt-3A expression was confined to the roof plate. In the developing brain Wnt-3 was expressed broadly across the dorsal portion of the neural tube with a rostral boundary of expression at the diencephalon. In contrast, Wnt-3A was expressed in a narrow region very close to the midline; expression extended into the bifurcating telencephalon, in a highly localized fashion. Both Wnt-3 and Wnt-3A were expressed in the ectoderm, and Wnt-3A was also expressed in the periumbilical mesenchyme. Characteristic expression patterns of these two closely related genes suggest that Wnt-3 and Wnt-3A play distinct roles in cell-cell signaling during morphogenesis of the developing neural tube.

Amino Acid Sequence

Temporal and spatial patterns of transin/stromelysin RNA expression following toxic injury in rat liver.

We have examined the expression of the extracellular matrix-degrading metalloprotease transin/stromelysin during the early phases of rat liver regeneration following toxic injury by a single dose of carbon tetrachloride (CCl4). In situ hybridization displayed cell type-specific spatial and temporal RNA expression patterns with high transcript levels in small proportions of hepatocytes and non-parenchymal cells, peaking at 24 and 48 h after intoxication, respectively. In agreement with the presence of c-fos and c-jun recognition sites on the transin gene, expression of these oncogenes preceded transin expression. Transin-expressing hepatocytes were largely localized in areas subsequently eliminated by necrosis due to CCl4 intoxication. As a consequence of these expression patterns and the key function of transin as an activator of interstitial collagenase, it seems that the hepatic fibrosis observed after CCl4 administration may be related to fibrogenesis unbalanced by fibrolysis due to altered transin expression.

Animals

Spatial patterns of hospital utilization: the impact of distance and time.

Although the impact of the physical proximity of health care facilities on utilization in rural areas is well established, its effect in metropolitan areas is still subject to question. This paper develops a spatial demand model of hospital choice to empirically estimate the impacts of distance and time on hospital utilization patterns. With a cross-product ratio estimation approach, the effects of physical access are estimated after controlling for spatial irregularities owing to the distribution of hospitals and population in metropolitan areas. The empirical results suggest that distance and time factors strongly influence hospital choice, even in metropolitan areas where alternatives are widely available, and that their effects vary across service classifications and hospitals.

Catchment Area, Health

Visual mechanisms for the analysis of spatial pattern.

After a brief outline of the structure and electrophysiology of the normal visual pathways, the responses, as revealed by psychophysical studies, of the visual system to spatially and temporally varying stimuli are reviewed. An appropriate network model, involving two sequentially organized classes of visual channel, is presented. Examples are given in which the psychophysical methods developed to analyse the normal visual pathways are applied to cases in which these pathways are defective (amblyopia, albinism, hemianopia, parietal cortical lesion, inhibitory central colour vision defect). These cases not only illustrate the value of psychophysical techniques in analysing disturbances of visual function but are also suggestive of the mechanisms involved in higher processing of the retinal image.

Adolescent

Minimum domains for spatial patterns in a class of reaction diffusion equations.

We study a general class of scalar reaction/interacting population diffusion equations in two space dimensions: convective terms, due to wind, are included. We consider boundary conditions which include a measure of the hostility to the species in the exterior of the domain. The main point of the paper is to obtain estimates for the minimum domain size which can sustain spatially heterogeneous structures and indicate the type of patterns which could appear.

Animals

Regulatory elements from the related spec genes of Strongylocentrotus purpuratus yield different spatial patterns with a lacZ reporter gene.

The Spec1 and Spec2 genes of Strongylocentrotus purpuratus are closely associated with the differentiation of aboral ectoderm. To examine cis-regulatory elements involved in the spatial expression of the Spec genes, we fused the Escherichia coli lacZ gene containing a nuclear targeting signal to 5'flanking DNA plus 5' untranslated leader sequences from Spec1, Spec2a, and Spec2c. All three genes contain 700 bp of highly conserved DNA in their upstream regions, but in Spec1 and Spec2c large insertions interrupt the conserved regions. The Spec-lacZ reporter gene plasmids were microinjected into eggs of S. purpuratus, Lytechinus variegatus, and L. pictus, and beta-galactosidase activity was determined in situ by X-gal staining. The Spec2a-lacZ fusion gene, which contained 1516 bp of 5' flanking DNA and 18 bp of 5' untranslated leader sequence, was preferentially expressed in aboral ectoderm cells in all three species. The Spec1-lacZ fusion gene was expressed in a strikingly different fashion--preferentially in primary and secondary mesenchyme cells, occasionally in aboral ectoderm cells, and less often in oral ectoderm and endoderm cells. The staining pattern was the same in either homologous or heterologous embryos. The Spec2c-lacZ fusion gene, like Spec2a-lacZ, was preferentially expressed in aboral ectoderm, but staining of other cell types was frequently observed. To further delineate sequences required for correct spatial expression, we deleted 800 bp of 5' flanking DNA from the Spec2a-lacZ fusion gene, resulting in a delta Spec2a-lacZ fusion gene that contained only the conserved DNA region. This gene fusion showed preferential expression in aboral ectoderm cells. However, the cell type specificity was not as great as with the parental Spec2a-lacZ plasmid. These experiments implied that the conserved DNA region, associated with all Spec genes examined, was insufficient for complete aboral ectoderm specificity, and suggested that a spatial repressor element existed between -1516 and -697 bp in the 5' flanking DNA of Spec2a.

Animals

Spatial patterns of gene expression in Brassica napus seedlings: identification of a cortex-specific gene and localization of mRNAs encoding isocitrate lyase and a polypeptide homologous to proteinases.

We investigated the spatial expression of three genes that are expressed during seed germination and postgerminative development in Brassica napus L. using in situ hybridization procedures. Two of the mRNAs encode isocitrate lyase and a predicted polypeptide that is homologous to cysteine proteinases. We reported previously that the mRNAs are prevalent primarily in cotyledons of seedlings and accumulate with similar kinetics during postgerminative growth. Here, we show that the two mRNAs are detected in several seedling tissues, but they display different distribution patterns in both cotyledons and root-shoot axes. The third mRNA is abundant in seedling axes and accumulates specifically in the ground meristem and mature cortex of hypocotyls and roots. Distribution of the mRNA in root meristems suggests that the gene product participates in an early event in cortical cell differentiation. Our results provide insight into the physiological processes that characterize seedlings.

Amino Acid Sequence