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QTL analysis of citrus tristeza virus-citradia interaction.

Citrus tristeza virus (CTV) has caused the death of millions of trees grafted on sour orange ( Citrus aurantium). However, this rootstock is very well adapted to the Mediterranean, semi-arid conditions. The aim of the present research is to genetically analyze the accumulation of CTV in a progeny derived from the cross between C. aurantium and Poncirus trifoliata, both resistant to CTV isolate T-346. Graft propagation of 104 hybrids was done on healthy sweet orange as a rootstock. Three months later, each rootstock was graft inoculated with two patches of infected tissue (isolate T-346). One, 2, and sometimes, 3 and 4 years after inoculation, hybrids and infected patches were tested for CTV by tissue-blot immuno-assay. Additionally, CTV multiplication was evaluated every year as the optical density of double-antibody sandwich enzyme-linked immuno-sorbent assay reactions. Linkage maps for P. trifoliata based on 63 markers, and for C. aurantium based on 157 markers, were used. Most molecular markers were microsatellites and IRAP (inter-retrotransposon amplified polymorphisms). Some analogues of resistance and expressed sequences were also included for candidate gene analysis. Resistance against CTV was analyzed as a quantitative trait (CTV accumulation) by QTL (quantitative trait loci) analysis to avoid the assumption of monogenic control. Three major resistance QTLs were detected where the P. trifoliata resistance gene, Ctv-R, had been previously located in other progenies. Up to five minor QTLs were detected ( Ctv-A(1) to Ctv-A(5)). A significant epistatic interaction involving Ctv-R(1) and Ctv-A(1) was also found. An analogue of a resistance gene is a candidate for Ctv-A(3), and two expressed sequences are candidates for Ctv-A(1) and Ctv-A(5). Single-strand conformational polymorphism analysis of CTV genes QTL P20 and P25 (coat protein) in susceptible hybrids, was carried out to test whether or not any QTL accumulation was a defeated resistance gene. Since the same haplotype of the virus was visualized independently on the CTV titer, differences in the amount of virions are not explained through the selection of CTV genotypes by the host, but through differences among citradias in CTV replication and/or movement.

Chromosome Mapping↗

Cytochrome b mutations in Leber hereditary optic neuropathy.

New mutations were discovered in the apocytochrome b gene in Leber hereditary optic neuropathy probands who did not harbor either of the two known Complex I mutations (positions 3,460 and 11,778). A mutation at position 15,257 was found in eight independent probands which changed a highly conserved aspartate to asparagine, was not found in controls, and appears to be pathogenetically significant. The 15,257 mutation occurred in association with a known synergistic mutation at position 13,708 in 7/8 probands and in association with a new apocytochrome b mutation at position 15,812 in 4/8 probands. Mutations in Complex III genes may be involved in Leber hereditary optic neuropathy and multiple, simultaneous mutations occur frequently.

Amino Acid Sequence↗

Lack of binocular activation of cells in area 19 of the Siamese cat.

Single cells were recorded in area 19 of 8 Siamese cats. Receptive fields (RFs) were typical for this area in terms of size, directional specificity and type. However, 69 out of the 70 units found were monocularly driven through the contralateral eye. Moreover, the amount of excursion of RFs into the ipsilateral visual field was more limited than that generally demonstrated for areas 17 and 18, extending to a maximum of 5 degrees with very few cells having RFs situated completely within the ipsilateral hemifield.

Animals↗

Functional anatomy of the tectum mesencephali of the goldfish. An explorative analysis of the functional implications of the laminar structural organization of the tectum.

The present paper is aimed at an exploration of the possible functional significance of the laminar organization of the goldfish tectum at both the cellular and the synaptic level. For this purpose (1) the data concerning the structure of the teleostean tectum are surveyed, (2) a conceptual framework of the intratectal connectivity in the goldfish is proposed, (3) the electrophysiological data concerning the teleosteam tectum are surveyed and (4) the degree of correlation between the structural and physiological data available is discussed. Apart from the retina, tectal afferents originate from at least 10 other brain centers. At least 5 of these projections appear to be topographically organized. Tectal afferents, neurons as well as synapses reveal a characteristic intratectal lamination pattern. Tectal efferents project to at least 10 brain centers, and have until now been shown to arise from 6 cell types. The structural data surveyed allow the construction of a conceptual framework of tectal circuitry on the basis of 3 starting points. (1) The existence of at least 8 presynaptic zones or laminae, each containing a characteristic set of presynaptic structures (afferents and axons of interneurons). (2) The fact that the tectal postsynaptic structures (somata and dendrites of tectal neurons) each have a characteristic location, extension and synaptic density, which determines the relative importance of the different presynaptic zones for each cell type. (3) The laminar specificity hypothesis, which implies that presynaptic structures that coexist in a particular presynaptic zone terminate without preference on all types of postsynaptic structures within that zone. The conceptual framework of tectal circuitry is quantified in terms of connectivity index and connective importance. Analysis of the framework constructed leads to a detailed description of the intratectal pathways involved in the processing of the 4 main streams of tectal input (i.e. visual, toral, telencephalic and 'deep' input). It was concluded that the laminar organization of the tectum is primarily relevant for multimodal integration and that the tectal cell types each receive a characteristic sample out of the multimodal information available in the different tectal layers.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

On the course and origin of cranial nerves in the teleost fish Gnathonemus determined by ortho- and retrograde horseradish peroxidase axonal transport.

By horseradish peroxidase (HRP) labelling the course and origin of the II-X cranial nerves are identified in the teleost fish Gnathonemus petersii. Roots as well as motor and sensory nuclei in the viscero- and somatomotor and somatosensory areas are localised. Earlier data of comparative and experimental anatomical observations are completed, partly confirmed or corrected.

Animals↗

Promotion of MAP/MAP interaction by taxol.

The effects of taxol on microtubule-associated proteins of high molecular weight (MAPs) were studied in vitro. After negative staining, microtubules reconstituted in the presence of taxol from preparations of partially purified tubulin and MAPs, besides being bundled, displayed prominent elongated or globular extensions without apparent regularity. These extensions, but not the tubulin polymer, were heavily decorated after immuno-gold-labeling using antibodies to MAP-1 and MAP-2. Microtubules reconsituted in the absence of taxol showed a much more regular, and apparently helical, arrangement of MAPs along their surfaces. The formation of polymeric structures was also observed when preparation of MAPs free of tubulin were incubated with taxol. In this case in addition to large network-type aggregates with little apparent substructure, more regular structures seemingly consisting of approximately 5-nm-thick filaments arrayed in parallel were observed. Taxol-induced MAP aggregation occurred rapidly and was directly proportional to the concentration of protein, as revealed by optical density measurements. It is concluded that taxol, aside from promoting the assembly of tubulin and stabilizing microtubules, promotes MAP/MAP interaction.

Alkaloids↗

Variable genotype of Leber's hereditary optic neuropathy patients.

Leber's hereditary optic neuropathy is caused by a single nucleotide change in the mitochondrial deoxyribonucleic acid (mtDNA). Each cell contains thousands of mitochondrial DNA molecules. We demonstrated that in certain isolated instances, the proband and close maternal lineage relatives can have mixtures of mutant and normal mitochondrial DNA molecules (heteroplasmy). The proportion of mutant mitochondrial DNA molecules was found to shift markedly across generations and within the tissues of an individual. One unaffected mother had 65% mutant mitochondrial DNA molecules whereas her affected son had essentially 100% mutant mitochondrial DNA molecules. Two affected individuals had predominantly mutant mitochondrial DNA in their blood, but significant normal mitochondrial DNA in their hair. The demonstration of heteroplasmy within maternal lineages and affected individuals means that the successful determination of the mitochondrial DNA genotype of a family or patient with Leber's hereditary optic neuropathy requires testing of more than one family member and more than one tissue from each individual.

Blotting, Southern↗

Heme structure and orientation in single monolayers of cytochrome c on polar and nonpolar soft surfaces.

Polarized x-ray absorption fine structure (XAFS) spectroscopy has been performed in fluorescence mode under total external reflection conditions on frozen hydrated single monolayers of yeast cytochrome c (YCC). The protein molecules were vectorially oriented within the monolayer by tethering their naturally occurring and unique surface cysteine residues to the sulfhydryl-endgroups at the surface of a mixed organic self-assembled monolayer, itself covalently attached to an ultrapure silicon wafer. The sulfhydryl-endgroups were isolated by dilution with either methyl- or hydroxyl-endgroups, producing macroscopically nonpolar or uncharged-polar soft surfaces, respectively. Independent information on the heme-plane orientation relative to the monolayer plane was obtained experimentally via optical linear dichroism. The polarized XAFS data have been analyzed both qualitatively and by a global mapping approach limited to systematically altering the various iron-ligand distances within a model for the local atomic environment of the heme prosthetic group, and comparing the theoretically generated XAFS spectra with those obtained experimentally. A similar analysis of unpolarized XAFS data from a frozen solution of YCC was performed using either the heme environment from the NMR solution or the x-ray crystallographic data for YCC as the model structure. All resulting iron-ligand distances were then used in molecular dynamics (MD) computer simulations of YCC in these three systems to investigate the possible effects of anisotropic ligand motions on the fits of the calculated to the experimental XAFS spectra.

Absorptiometry, Photon↗

Activation of color-selective areas of the visual cortex in a blind synesthete.

Many areas of the visual cortex are activated when blind people are stimulated naturally through other sensory modalities (e.g., haptically; Sadato et al., 1996). While this extraneous activation of visual areas via other senses in normal blind people might have functional value (Kauffman et al., 2002; Lessard et al., 1998), it does not lead to conscious visual experiences. On the other hand, electrical stimulation of the primary visual cortex in the blind does produce illusory visual phosphenes (Brindley and Lewin, 1968). Here we provide the first evidence that high-level visual areas not only retain their specificity for particular visual characteristics in people who have been blind for long periods, but that activation of these areas can lead to visual sensations. We used fMRI to demonstrate activity in visual cortical areas specifically related to illusory colored and spatially located visual percepts in a synesthetic man who has been completely blind for 10 years. No such differential activations were seen in late-blind or sighted non-synesthetic controls; neither were these areas activated during color-imagery in the late-blind synesthete, implying that this subject's synesthesia is truly a perceptual experience.

Association↗

Axon-glia interactions and the domain organization of myelinated axons requires neurexin IV/Caspr/Paranodin.

Myelinated fibers are organized into distinct domains that are necessary for saltatory conduction. These domains include the nodes of Ranvier and the flanking paranodal regions where glial cells closely appose and form specialized septate-like junctions with axons. These junctions contain a Drosophila Neurexin IV-related protein, Caspr/Paranodin (NCP1). Mice that lack NCP1 exhibit tremor, ataxia, and significant motor paresis. In the absence of NCP1, normal paranodal junctions fail to form, and the organization of the paranodal loops is disrupted. Contactin is undetectable in the paranodes, and K(+) channels are displaced from the juxtaparanodal into the paranodal domains. Loss of NCP1 also results in a severe decrease in peripheral nerve conduction velocity. These results show a critical role for NCP1 in the delineation of specific axonal domains and the axon-glia interactions required for normal saltatory conduction.

Aging↗

Cortical activation to illusory shapes as measured with magnetoencephalography.

Spatiotemporal patterns of cortical activation during the perceptual grouping of elements to form illusory shapes were estimated using anatomically constrained magnetoencephalography. Subjects were shown an array of Kanizsa-style figures which were either aligned to form illusory squares or misaligned so that no illusory contour or shape was perceived. Differential activity is more pronounced in the right hemisphere. After a weakly significant modulation at approximately 110 ms in the occipital pole, a prominent peak appears at approximately 155 ms in the lateral occipital cortex. Modulation then appears to spread back from this location toward the occipital pole, as well as ventrally to involve ventral occipital and temporal cortices for the next 180 ms, eventually involving ventral orbitofrontal cortex at 325 ms. The prominent lateral occipital response is consistent with fMRI studies with similar stimuli which found activation in that region as well as in V3A, V4v, V7, and V8. Furthermore, the timing of this activation, after the occipital pole but before ventral temporal, is consistent with a putative role for this region in midlevel vision. The late ventral temporal response (235 ms) is centered in the lingual and fusiform areas implicated in object identification. The V1/V2 modulation at this time may reflect top-down modulation by lateral occipitotemporal and ventral temporal areas.

Adult↗

Visual motion aftereffect in human cortical area MT revealed by functional magnetic resonance imaging.

Functional magnetic resonance imaging (fMRI) was used to measure local haemodynamic changes (reflecting electrical activity) in human visual cortex during production of the visual motion aftereffect, also known as the waterfall illusion. As in previous studies, human cortical area MT (V5) responded much better to moving than to stationary visual stimuli. Here we demonstrate a clear increase in activity in MT when subjects viewed a stationary stimulus undergoing illusory motion, following adaptation to stimuli moving in a single local direction. Control stimuli moving in reversing, opposed directions produced neither a perceptual motion aftereffect nor elevated fMRI levels postadaptation. The time course of the motion aftereffect (measured in parallel psychophysical tests) was essentially identical to the time course of the fMRI motion aftereffect. Because the motion aftereffect is direction specific, this indicates that cells in human area MT are also direction specific. In five other retinotopically defined cortical areas, similar motion-specific aftereffects were smaller than those in MT or absent.

Afterimage↗

A mitochondrial DNA mutation at nucleotide pair 14459 of the NADH dehydrogenase subunit 6 gene associated with maternally inherited Leber hereditary optic neuropathy and dystonia.

A five-generation Hispanic family expressing maternally transmitted Leber hereditary optic neuropathy and/or early-onset dystonia associated with bilateral basal ganglia lesions was studied. Buffy coat mitochondrial DNA (mtDNA) from a severely affected child was amplified by the polymerase chain reaction and greater than 90% sequenced. The mtDNA proved to be a Native American haplogroup D genotype and differed from the standard "Cambridge" sequence at 40 nucleotide positions. One of these variants, a G-to-A transition at nucleotide pair (np) 14459, changed a moderately conserved alanine to a valine at NADH dehydrogenase subunit 6 (ND6) residue 72. The np 14459 variant was not found in any of 38 Native American haplogroup D mtDNAs, nor was it detected in 108 Asian, 103 Caucasian, or 99 African mtDNAs. Six maternal relatives in three generations were tested and were found to harbor the mutation, with one female affected with Leber hereditary optic neuropathy being heteroplasmic. Thus, the np 14459 G-to-A missense mutation is specific to this family, alters a moderately conserved amino acid in a complex I gene, is a unique mtDNA variant in Native American haplogroup D, and is heteroplasmic, suggesting that it is the disease-causing mutation.

Adenine↗

Magnetically induced phosphenes in sighted, blind and blindsighted observers.

Direct stimulation of visual cortex can produce illusory flashes of light, called phosphenes. Here we describe the spatial and motion properties of phosphenes produced by transcranial magnetic stimulation in normal subjects and in two subjects with peripheral or cortical blindness. The totally retinally blind subject experienced normal phosphenes, apart from their concentration in the centre of the visual field, whereas the hemianopic subject, lacking area V1, did not experience phosphenes when his surviving extrastriate visual areas were stimulated. In the absence of V1, magnetically induced activity was unable to generate a conscious visual percept in the field defect.

Adult↗