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Binding of phospholipase A2 to zwitterionic bilayers is promoted by lateral segregation of anionic amphiphiles.

Catalytic action of phospholipase A2 is appreciably influenced by the organization and dynamics of bilayers of glycerophosphocholines (Apitz-Castro et al. (1988) Biochim. Biophys. Acta 688, 341-348). However, such effects of the quality of the interface are not observed with bilayers of glycerophosphoryl methanol and other anionic phospholipids (Jain et al. (1986) Biochim. Biophys. Acta 860, 435-447). Such differences between the catalytic susceptibility of zwitterionic versus anionic bilayers are due to a large difference in the affinity of the enzyme for these interfaces. Binding to phospholipase A2 to zwitterionic interfaces can be promoted in the presence of certain anionic additives. For example in the pre-steady-state phase of hydrolysis, segregation of the nacently produced products of hydrolysis could promote binding of phospholipase A2 to regions of higher anionic charge density in the zwitterionic interface. In this paper we show that the dynamics of segregation of the nacently produced products of hydrolysis in zwitterionic bilayers can be readily followed by monitoring the fluorescence intensity of the cationic dye NK-529 (Yu and Jain (1989) Biochim. Biophys. Acta 980, 15-22). The fluorescence emission characteristics of NK-529 change appreciably due to self-quenching of the bound dye molecules as the fatty acid molecules segregate in the bilayer. The kinetics of segregation of fatty acids during the course of hydrolysis of bilayers of zwitterionic phospholipids by phospholipase A2 exhibits an unequivocal correlation with a variety of phenomena that are observed during the transition from the pre-steady-state phase to the steady-state phase of hydrolysis in the reaction progress curves as a function of temperature and in the presence of lipophilic additives.

Anions↗

Ultraviolet irradiation during ooplasmic segregation prevents gastrulation, sensory cell induction, and axis formation in the ascidian embryo.

The effect of ultraviolet (uv) light on embryonic development was examined in the ascidian Styela clava. uv irradiation (3.0 x 10(-3) J mm-2) of the entire surface of fertilized eggs during ooplasmic segregation prevented gastrulation, sensory cell induction, and embryonic axis formation. The uv-irradiated embryos completed ooplasmic segregation and cleaved normally, but vegetal blastomeres did not invaginate at the beginning of gastrulation, sensory cells in the larval brain did not develop tyrosinase or melanin pigment, and the larval tail did not develop. Endoderm, epidermis, and muscle cells differentiated in the uv-irradiated embryos, however, as evidenced by expression of endodermal alkaline phosphatase (AP), an epidermal-specific antigen, and alpha-actin, myosin heavy chain, and acetylcholinesterase (AChE) in muscle cells. Higher doses of uv light (6.0-9.0 x 10(-3) J mm-2) suppressed expression of the epidermal antigen and muscle cell markers, whereas the development of endodermal AP was insensitive. Irradiation at various times between fertilization and the 16-cell stage revealed that gastrulation, sensory cell differentiation, and axis formation are sensitive to uv light only during ooplasmic segregation. Irradiation of restricted regions of the zygote during ooplasmic segregation showed that the uv-sensitive components are localized in the vegetal hemisphere. The absorption characteristics of the uv-sensitive components suggest that they are nucleic acids. The results show that uv-sensitive components that specify gastrulation, sensory cell induction, and embryonic axis formation are localized in the vegetal hemisphere of Styela eggs.

Acetylcholinesterase↗

The effect of bromodeoxyuridine on the segregation of the chicken-specific HPRT gene from Chinese hamster-chick red blood cell somatic hybrids.

The segregation of the chick-specific HPRT gene was studied in three Chinese hamster-chick red blood cell hybrid lines. The three lines showed individual segregation kinetics, the segregation taking place in an exponential-like fashion. Bromodeoxyuridine becomes incorporated into the nuclear DNA and increases the spontaneous segregation rate.

Animals↗

Evolution of the segregation ratio: modification of gene conversion and meiotic drive.

We compare the evolutionary pressures that direct the modification of gene conversion and meiotic drive at loci subject to purifying and overdominant viability selection. Gene conversion differs from meiotic drive in that modifers do not affect their own segregation ratios, even when linked to the viability locus. Segregation distortion generates gametic level disequilibria between alleles at the viability locus and modifiers of gene conversion and meiotic drive: enhancers of segregation distortion become positively associated with driven alleles. Suppression of gene conversion evolves if the driven allele is marginally disadvantageous (overdominant viability selection), and higher rates evolve if the driven alleles are relatively advantageous (purifying viability selection). Gametic disequilibria permit enhancers of meiotic drive that are linked to the driven locus to promote their own segregation. We attribute the failure of genetic modifiers of gene conversion and meiotic drive to maximinize mean fitness to the generation of such associations.

Biological Evolution↗

Electrophysiological correlates of texture segregation in the human visual evoked potential.

We investigated whether the visual evoked potential (VEP) reflects cortical processing associated with preattentive texture segregation. On a visual display unit we presented stimuli with various arrangements of oriented line segments that either led to the appearance of a "preattentive" checkerboard or did not. Two presentation modes were used (pattern onset at 1 Hz and rapid pattern change at 4.3 Hz), while luminance (57 cd/m2) and contrast (92%) of the line segments remained constant. VEPs were recorded in 7 human subjects. The VEP was analyzed as a linear combination of putative components, which are evoked by either local pattern, quasi-local orientation contrast or global preattentive structure. In the transient VEP, we found a negativity over the posterior pole at a latency between 161 and 225 msec (FWHM) in the linear combination designed to extract segregation-specific components. Peak amplitude reached 3.1 +/- 0.8 microV (mean +/- SEM) at 199 msec. This negative peak appeared only for textures containing orientation contrast. Steady-state analysis of the rapid presentation also revealed a significant component (P = 0.002) associated with texture segregation. These potentials either represent processing of orientation contrast or global processing of texture segregation. The results suggest that specific surface potentials, differing from cognitive potentials, can be derived which are associated with preattentive processing.

Adult↗

Functional segregation of color and motion perception examined in motion nulling.

We examine two hypotheses about the functional segregation of color and motion perception, using a motion nulling task. The most common interpretation of functional segregation, that motion perception depends only on one of the three dimensions of color, is rejected. We propose and test an alternative formulation of functional segregation: that motion perception depends on a univariate motion signal driven by all three color dimensions, and that the motion signal is determined by the product of the stimulus contrast and a term that depends only on the relative cone excitations. Two predictions of this model are confirmed. First, motion nulling is transitive: when two stimuli null a third they also null another. Second, motion nulling is homogeneous: if two stimuli null one another, they continue to null one another when their contrasts are scaled equally. We describe how to apply our formulation of functional segregation to other behavioral and physiological measurements.

Color Perception↗

Spatial-frequency- and orientation-selectivity of simple and complex channels in region segregation.

Models incorporating spatial-frequency- and orientation-selective channels explain many texture-segregation results, particularly when known nonlinearities are included. One such nonlinearity is complex channels. A complex channel consists of two stages of linear filtering separated by a rectification-type nonlinearity. Here we investigate the spatial-frequency- and orientation-selectivity of simple (linear) channels and of the complex channels' first stage. Observers rated the degree of segregation between two "textures" both composed of elements which were Gabor patches. When the textures differed in type of element (e.g. one composed of vertical and the other of horizontal Gabor patches), the segregation results yield bandwidth estimates for simple channels of approx. 0.5-1.0 octave on the spatial-frequency dimension and 5-20 deg of rotation on the orientation dimension. When the textures differ in the arrangement of elements (e.g. striped vs checkerboard arrangements, both of horizontal and vertical patches), the segregation results yield bandwidth estimates for the first stage of complex channels. These estimates, while differing substantially from one observer to another, were always substantially wider than those for simple channels (by at least a factor of two) but narrower than bandwidths of LGN cells (particularly on the orientation dimension where LGN cells show little selectivity at all).

Contrast Sensitivity↗

Evidence for two speed signals: a coarse local signal for segregation and a precise global signal for discrimination.

In calculating the precise speed of an object, the visual system must integrate motion measurements across time and space while keeping motion measurements from different objects separate. We examined whether an initial coarse estimate of local speed may be used to segregate the motions of different objects prior to a precise calculation of object speed. Our stimuli consisted of 256 dots that moved upward at two speeds. In Expt 1, each dot alternated between the two speeds every 133 msec. When the speed alternations were asynchronous across dots, subject saw two transparent surfaces moving at different speeds and their ability to discriminate changes in the slow speed were unaffected by the presence of the fast speed. This experiment suggests that before integration, motion measurements may be segregated according to speed. We sought more conclusive evidence for this claim in Expts 2 and 3. In Expt 2, dots with 33 msec lifetimes were used to generate the two speeds. Although individual dots permitted only crude speed discrimination, subjects perceived this stimulus as two surfaces moving at different speeds and they precisely judged the slower speed. Apparently, the coarse local signals generated by the slow dots were segregated from those of the fast dots and then separately integrated to produce a precise speed signal. In Expt 3, the dots again moved at two speeds, but each speed was generated by a range of spatial and temporal displacements. Once more, subjects saw two surfaces and precisely judged the speed of the slower surface, demonstrating that segregation may be based solely on differences in local speed.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential Threshold↗

Investigating simple and complex mechanisms in texture segregation using the speed-accuracy tradeoff method.

Several recent models of texture segregation have proposed two mechanisms: simple, linear channels (first-order, Fourier mechanisms) and complex channels (second-order, non-Fourier mechanisms). We used the speed-accuracy tradeoff (SAT) method to examine the time course of texture segregation processing in simple and complex channels. The stimuli were texture patterns designed to segregate primarily as a result of activity in one set of channels or the other. We presented subjects with textures that were checked or striped arrangements of either Gaussian-blob or Gabor-patch elements. Subjects were required to identify the orientation of a rectangular texture region embedded in a background field of a different texture. A range of contrasts and a control task were used to equate visibility of the Gabor and Gaussian textures. SAT functions were obtained by requiring subjects to respond within 200 msec after an auditory cue. We found that when segregation depended primarily on simple channels, performance was faster than when it depended primarily on complex channels: the 75% correct level was reached 100-200 msec sooner and this extra speed was reflected both in smaller delay and higher rate parameters.

Contrast Sensitivity↗

The asymmetric segregation of parental nucleosomes during chrosome replication.

SV40 DNA replicated in the presence of cycloheximide was more sensitive to staphylococcal nuclease digestion and had a lower superhelical density than viral DNA replicated in the absence of this drug. These data indicate that fewer nucleosomes are associated with progeny SV40 DNA molecules after DNA replication in the absence of protein synthesis and that these nucleosomes are derived from the parental histones. We designed an experiment to determine whether these parental SV40 nucleosomes segregate to the leading side of the replication form where DNA synthesis is continuous, the lagging side of the fork where DNA synthesis is discontinuous or randomly to both sides of the fork. The results indicate that the parental histones distributed themselves asymmetrically, preferentially (80-90%) segregating with the leading side of both SV40 DNA replication forks during bidirectional replication in the absence of protein synthesis. In the case of SV40, the same parental DNA strands are the templates for the leading side of DNA replication at both forks as well as the templates for the informational or coding strand of early and late viral mRNA synthesis. Based on this correspondence, we designed an experiment to test whether chicken cells growing in culture and replicating their DNA in the absence of protein synthesis segregated their parental histones asymmetrically to the progeny DNA strand that also coded for stable nuclear RNA transcripts. The results of these experiments indicate that, like SV40, parental cellular histones segregate asymmetrically and are preferentially associated with those DNA template strands that code for stable nuclear RNA species detected by hybridization to single-copy DNA.

Cycloheximide↗

Chromosome length controls mitotic chromosome segregation in yeast.

We have examined the effect of physical length on the mitotic segregation of artificial chromosomes and fragments of natural yeast chromosomes. Increasing the length of artificial chromosomes decreases the rate at which they are lost during mitosis. We have made fragments of chromosome III by integrating new telomeres at different positions along the length of the chromosome. Chromosome fragments of 42 and 72 kb behave like artificial chromosomes: they are lost in mitosis much more frequently than natural chromosomes. In contrast, a chromosome fragment of 150 kb is as mitotically stable as the full-length chromosome from which it is derived. The structural instability of a short dicentric artificial chromosome demonstrates that, although short artificial chromosomes segregate poorly in mitosis, they do attach to the mitotic spindle. We discuss these results in the context of a model in which chromosome segregation is directed by the intercatenation of the segregating DNA molecules.

Chromosomes↗

Characterization of a region of the Enterococcus faecalis plasmid pAM beta 1 which enhances the segregational stability of pAM beta 1-derived cloning vectors in Bacillus subtilis.

The nucleotide sequence of a 2.13-kb EcoRI-HindIII, pAM beta 1-derived fragment, isolated from the gram-positive cloning vector pHV1431, has been determined and shown to encode two ORFs. ORF H encodes for a protein of 23,930 Da which exhibits substantial homology to bacterial site-specific recombinases, particularly the resolvases of the gram-positive transposons Tn917 (30.3% identity) and Tn552 (31.6% identity) and the clostridial plasmid pIP404 (27.1% identity). The second ORF (I) is incomplete and encodes a polypeptide which has significant homology with Escherichia coli topoisomerase I (26.0% identity). Insertion of either the entire 2.13-kb EcoRI-HindIII fragment or a 0.73-kb EcoRI-DraI subfragment encoding only the resolvase into the pAM beta 1-based cloning vector pMTL500E causes a significant enhancement of segregational stability (from 6.5 X 10(-2) to 3.0-4.0 X 10(-3) plasmid loss per cell per generation). Improved segregational stability is mirrored by a reduction in plasmid polymerization. The introduction of a stop codon into the resolvase coding region negates its ability to promote segregational stability. It is proposed that the identified determinant stabilizes pAM beta 1-based vectors in Bacillus subtilis by maintaining the plasmid population in the monomeric state, thereby reducing the chances of producing plasmid-free segregants.

Amino Acid Sequence↗

Development of abnormal lamination and binocular segregation in the retinotectal pathways of the rat.

The uncrossed retinotectal pathway of pigmented rats originates from a small fraction of the retinal ganglion cell population. This projection terminates deeply in discrete patches within the upper grey layers where crossed and uncrossed inputs overlap. However, after the experimental enlargement of the uncrossed pathway, the ipsilateral fibers are also found in a superficial tier of the upper grey layers where binocular inputs segregate [36]. We studied the development of the retinotectal projections in rats after the enlargement of the uncrossed pathway as a result of a contralateral (left) optic tract lesion (OTL) made at birth. Horseradish peroxidase (HRP) was used as an anterograde tracer. An abnormal uncrossed projection from the right eye to the collicular surface appeared at postnatal day 3 (P3). Between P5 and P10, this projection developed the bilaminar pattern seen in similar operated adults. The laminar arrangement of the aberrant terminal fields did not change significantly after an ipsilateral visual cortex ablation on the day of birth. Despite the early development of the aberrant uncrossed pathway, binocular segregation was incipient at P10. At P14, 46% of the operated rats presented gaps in the terminal labeling at the tectal surface. This figure increased to 55.5% at 6 weeks, a proportion still smaller than in adult animals of the same group (69%). Eyelid suture had no effect on segregation. This projection remains plastic for at least 3 weeks, since the removal of the ipsilateral input at either P14 or P21 resulted in the absence of gaps in the contralateral projection. We conclude that the laminar selection of retinotectal projections depends on binocular interactions and that the abnormal segregation of retinal inputs to the superior colliculus has an unusually protracted development which can be reversed long after the previously defined critical period in this system.

Aging↗

Target-dependent morphological segregation of Aplysia sensory outgrowth in vitro.

The adult nervous system is characterized by partial or complete morphological segregation of terminals from different afferent neurons innervating the same postsynaptic target. This segregation is thought to result, in part, from competition between the afferent terminals. To explore the role of the target cell in the spatial distribution of presynaptic inputs, the sensory neurons of Aplysia were cultured either with or without a common target motor neuron. In the presence of a common target, the outgrowth from two different sensory neurons tends to occupy separate postsynaptic regions. When cultured without a target motor neuron, processes from different sensory neurons do not segregate, but rather grow freely along one another. Thus, morphological segregation of sensory outgrowth requires interaction with a target neuron and may reflect competition between presynaptic terminals for a limited number of synaptic sites on the motor neuron, or for a postsynaptic trophic factor.

Afferent Pathways↗

Analyzing of segregation in mixtures of 3-methylpyridine and heavy water by dynamic neutron radiography.

The closed-loop phase diagram of 3-methylpyridine-heavy water mixture was studied, to our best knowledge, for the first time with dynamic neutron radiography (DNR) at the 10MW VVR-SM Research Reactor in Budapest (Hungary). Visualized were the (i) lower temperature non-segregated states (below 38 degrees C), (ii) transition (40-43 degrees C), (iii) segregated states (46-128 degrees C), (iv) higher temperature transition (110-128 degrees C) and (v) higher temperature non-segregated states. The non-segregated state belonging to 141 degrees C, was found to be definitely dissimilar from the lower temperature state. Existence of a solid-like structure at the liquid-liquid interface was indicated.

Journal Article↗

Somatotopy in the basal ganglia: experimental and clinical evidence for segregated sensorimotor channels.

Growing experimental and clinical evidence supports the notion that the cortico-basal ganglia-thalamo-cortical loops proceed along parallel circuits linking cortical and subcortical regions subserving the processing of sensorimotor, associative and affective tasks. In particular, there is evidence that a strict topographic segregation is maintained during the processing of sensorimotor information flowing from cortical motor areas to the sensorimotor areas of the basal ganglia. The output from the basal ganglia to the motor thalamus, which projects back to neocortical motor areas, is also organized into topographically segregated channels. This high degree of topographic segregation is demonstrated by the presence of a well-defined somatotopic organization in the sensorimotor areas of the basal ganglia. The presence of body maps in the basal ganglia has become clinically relevant with the increasing use of surgical procedures, such as lesioning or deep brain stimulation, which are selectively aimed at restricted subcortical targets in the sensorimotor loop such as the subthalamic nucleus (STN) or the globus pallidus pars interna (GPi). The ability to ameliorate the motor control dysfunction without producing side effects related to interference with non-motor circuits subserving associative or affective processing requires the ability to target subcortical areas particularly involved in sensorimotor processing (currently achieved only by careful intraoperative microelectrode mapping). The goal of this article is to review current knowledge about the somatotopic segregation of basal ganglia sensorimotor areas and outline in detail what is known about their body maps.

Animals↗

Infants' formation and use of categories to segregate objects.

Four- and-a-half-month-old infants' (N = 100) category formation and use was studied in a series of five experiments. For each experiment, the test events featured a display composed of a cylinder and a box. Previous research showed that this display is not clearly parsed as a single unit or as two separate units by infants of this age. Immediately prior to testing, infants were shown a set of category exemplars. Knowledge about this category could help infants disambiguate the test display, which contained a novel exemplar of this category. Clear interpretation of the test display as composed of two separate units (as indicated by infants' longer looking at the move-together than at the move-apart test event) was taken as evidence of category formation and use. In Experiments 1 and 5, infants' prior experience with a set of three different boxes that were similar to the test box facilitated their segregation of the test display. Experiment 2 showed that three different exemplars are necessary: prior experience with any two of the three boxes used in Experiment 1 did not facilitate infants' segregation of the test display. Experiment 3 showed that variability in the exemplar set is necessary: prior experience with three identical boxes did not facilitate infants' segregation of the test display. Experiment 4 showed that under these conditions of very brief prior exposure, similarity between the exemplar set and test box is necessary: prior experience with three different boxes that were not very similar to the test box did not facilitate infants' segregation of the test display. Together, these findings suggest that: (a) number of exemplars, variability, and similarity in the exemplar set are important for infants' category formation, and (b) infants use their category knowledge to determine the boundaries of the objects in a display.

Cognition↗

Effects of differences in the pattern of amplitude envelopes across harmonics on auditory stream segregation.

When a mixture of sound from many sources arrives at the ear, the auditory system attempts to segregate it into different perceptual streams. For sequences of sounds, the effects of basic acoustic properties (e.g., frequency separation, rate) on streaming are well understood, but much less is known about the effects of more complex acoustic attributes. Dynamic variations in frequency spectrum are known to have an important effect on the timbre of sounds. We investigated whether dynamic variations also affect stream segregation. Periodic tones were used, comprising harmonics 1-6, and presented in long sequences (ABA-ABA- ...). Tones A and B always differed in fundamental frequency, a factor known to influence streaming, and had either the same or different patterns of spectral variation. The total amount of variation (spectral flux) was held constant. Listeners judged moment-by-moment the grouping of these sequences, and the measure used was the proportion of time a sequence was heard as segregated. We found that sequences of complex tones with different patterns of spectral variation are more likely to segregate when this results in different patterns of change in the frequency centroid over time (Experiments 1 and 2), but not when it does not (Experiment 3).

Acoustic Stimulation↗