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Cloning and characterization of Tag 1, a tobacco anther beta-1,3-glucanase expressed during tetrad dissolution.

A critical stage in pollen development is the dissolution of the four products of meiosis, the tetrads, into free microspores. The tetrads are surrounded by a thick callose wall composed of beta-1,3-glucan. At the completion of meiosis, the tetrads are released into the anther locule after hydrolysis of the callose by a beta-1,3-glucanase. Using the polymerase chain reaction, we have amplified and subsequently cloned a cDNA corresponding to a beta-1,3-glucanase, tobacco (Nicotiana tabacum cv. Samsun) anther glucanase (Tag 1), which is expressed exclusively in anthers from meiosis to the free microspore stage of pollen development. The identity of the clone was determined by DNA and deduced protein sequence similarity to other known beta-1,3-glucanases. Several regions strictly conserved among four classes of glucanases are also conserved in the Tag 1 protein. Tag 1 represents a novel class of beta-1,3-glucanase based on phylogenetic analysis and RNA expression pattern. Tag 1 RNA was detected in situ only in the tapetum, with maximal expression just prior to tetrad dissolution. Due to its expression pattern and sequence similarity to other beta-1,3-glucanases, we believe Tag 1 may be involved in tetrad dissolution.

Amino Acid Sequence↗

Coordination between posture and movement in a bimanual load lifting task: putative role of a medial frontal region including the supplementary motor area.

The aim of the present experimental series was to investigate the role of the medial frontal region including the supplementary motor area in the coordination between posture and movement in a bimanual load lifting task. The seated subject was instructed to maintain in a horizontal position one forearm (postural arm) which was loaded with a 1 kg weight. The unloading was performed either by the experimenter (imposed unloading) or by a voluntary movement of the other arm (voluntary unloading). In normal individuals, with the voluntary unloading, the movement control was accompanied by an anticipatory adjustment of the postural forearm flexor activity, which resulted in the maintenance of the forearm position despite the unloading. The anticipatory postural adjustments were impaired in 4 out of 5 patients with unilateral lesion of the SMA region; the defect was observed mainly when the postural forearm was contralateral to the lesion. No change in the anticipatory postural adjustment was observed in one patient with complete callosal section. This finding indicates that the coordination between the posture and movement in this task is not organized through callosal fibers linking the cortices on both sides but rather at a subcortical level. The anticipatory postural adjustments were abolished in two patients with spastic hemiparesis when the postural forearm was the spastic arm. It is suggested that the SMA region contralateral to the postural forearm, together with other premotor or motor areas, may select the circuits responsible for the phasic postural adjustments which are necessary to ensure postural maintenance, whereas the motor cortex contralateral to the voluntary movement controls both the movement and, via collaterals, the preselected circuits responsible for the associated postural adjustments.

Adult↗

Binocular interaction and disparity coding at the 17-18 border: contribution of the corpus callosum.

Binocular disparity, resulting from the projection of a three-dimensional object on the two spatially separated retinae, constitutes one of the fundamental cues for stereoscopic perception. The binocularity of cells in one hemisphere stems from two sources: i) from the ipsilateral ganglion cells in the temporal retina which converge with inputs coming from the contralateral nasal retina; the latter axons cross at the chiasma; ii) from inputs originating in the opposite hemisphere which cross in the corpus callosum. The objective of this study was to demonstrate that interactions from both types of inputs can result in the formation of disparity sensitive neurons and presumably that either type could mediate stereoperception based on disparity cues. Two types of disparity sensitive neurons were found in the normal cat: one type, showing maximal interactive effects around zero disparity responded with strong excitation or inhibition when the stimuli were in register. These neurons are presumed to signal stimuli situated about the fixation plane. The other type, also made up of two subtypes of opposed valencies, gave maximum responses at one set of disparities and inhibitory responses to the other set. These were presumed to signal stimuli situated in front of or behind the fixation plane. In the split-chiasm cat, whose cortical binocularity is presumably assured by converging ipsilateral and callosal inputs, three of the four subtypes of disparity sensitive neurons were found, the uncrossed disparity cells being absent in these animals. Moreover, stimulating each eye individually indicated that nearly 80% of the cells in normal and about 40% in split-chiasm cats were binocularly driven. However, both these figures underestimated the amount of binocular interaction in the callosal recipient zone, since stimulating both eyes simultaneously showed that a proportionately larger number of cells were binocularly driven. Disparity sensitive cells also varied as a function of ocular dominance, i.e., cells signaling the fixation plane tended to have balanced dominance whereas units preferring stimuli situated in front of or behind the fixation plane were dominated by the ipsilateral and contralateral eyes, respectively.

Animals↗

Auditory corticocortical interconnections in the cat: evidence for parallel and hierarchical arrangement of the auditory cortical areas.

The origin and laminar arrangement of the homolateral and callosal projections to the anterior (AAF), primary (AI), posterior (PAF) and secondary (AII) auditory cortical areas were studied in the cat by means of electrophysiological recording and WGA-HRP tracing techniques. The transcallosal projections to AAF, AI, PAF and AII were principally homotypic since the major source of input was their corresponding area in the contralateral cortex. Heterotypic transcallosal projections to AAF and AI were seen, originating from the contralateral AI and AAF, respectively. PAF received heterotypic commissural projections from the opposite ventroposterior auditory cortical field (VPAF). Heterotypic callosal inputs to AII were rare, originating from AAF and AI. The neurons of origin of the transcallosal connections were located mainly in layers II and III (70-92%), and less frequently in deep layers (V and VI, 8-30%). Single unit recordings provided evidence that both homotypic and heterotypic transcallosal projections connect corresponding frequency regions of the two hemispheres. The regional distribution of the anterogradely labeled terminals indicated that the homotypic and heterotypic auditory transcallosal projections are reciprocal. The present data suggest that the transcallosal auditory interconnections are segregated in 3 major parallel components (AAF-AI, PAF-VPAF and AII), maintaining a segregation between parallel functional channels already established for the thalamocortical auditory interconnections. For the intrahemispheric connections, the analysis of the retrograde tracing data revealed that AAF and AI receive projections from the homolateral cortical areas PAF, VPAF and AII, whose neurons of origin were located mainly in their deep (V and VI) cortical layers. The reciprocal interconnections between the homolateral AAF and AI did not show a preferential laminar arrangement since the neurons of origin were distributed almost evenly in both superficial (II and III) and deep (V and VI) cortical layers. On the contrary, PAF received inputs from the homolateral cortical fields AAF, AI, AII and VPAF, originating predominantly from their superficial (II and III) layers. The homolateral projections reaching AII originated mainly from the superficial layers of AAF and AI, but from the deep layers of VPAF and PAF. The laminar distribution of anterogradely labeled terminal fields, when they were dense enough for a confident identification, was systematically related to the laminar arrangement of neurons of origin of the reciprocal projection: a projection originating from deep layers was associated with a reciprocal projection terminating mainly in layer IV, whereas a projection originating from superficial layers was associated with a reciprocal projection terminating predominantly outside layer IV.(ABSTRACT TRUNCATED AT 400 WORDS)

Acoustic Stimulation↗

The critical period for corpus callosum section to affect cortical binocularity.

The period of time during which surgical section of the corpus callosum (CC) is effective in altering the physiological properties of cells in cat striate cortex was investigated. Cats which had the CC transected between 13 days and 24 weeks of age were studied using extracellular, single-unit recording procedures. Analysis of the results from 1,747 cortical units indicate that when the CC was sectioned prior to 19 days of age there was a reduction in the encounter rate of binocularly activated neurons and an increase in the proportion of neurons dominated by the contralateral eye. The decrease in cortical binocularity was observed in both simple and complex cell populations, and at all receptive field eccentricities studied (0-39 degrees). However, when the CC was sectioned after 19 postnatal days, no physiological changes were detected. Thus, in contrast with previous studies (Payne et al. 1980a, b) no changes were found following CC section in adult cats. The results therefore define a critical period which ends before 3 weeks of age during which corpus callosum section reduces striate cortex binocularity. Although the corpus callosum critical period is much shorter than the critical period for experiential alterations in cortical binocularity, the physiologically determined limits of the callosal critical period agree with the behaviorally determined limits previously found for the callosal critical period (Elberger 1984).

Animals↗

Properties of visual cortical cells of the intact and the deafferented hemisphere of unilateral optic tract sectioned acute and chronic adult cats.

Single unit recording from visual cortex areas 17 and 18 and in the border region between them was performed on adult cats with unilateral optic tract section (OTX) either on the day of the operation (acute) or three to six months postoperatively (chronic). Visual activity from both hemispheres was analyzed with respect to the responsiveness level, the ocular dominance distribution and selectivity to orientation and direction. The results showed almost a complete absence of visual responsiveness in the deafferented hemisphere and a considerable reduction of responsiveness in the intact hemisphere. Following surgery an increase in visually responsive cells was found in the intact hemisphere as postoperative recovery continued. In addition, a reduction in the proportion of cells selective for orientation and direction was also found in the intact hemisphere of the OTX animals as compared to the control cats. Furthermore, cortical binocularity was not affected in the intact hemisphere of all OTX cats. We conclude that an almost total absence of interhemispheric callosal transfer of visual functions from the intact to the deafferented hemisphere is induced as a result of the unilateral OTX in adult cats. Moreover, the fact that the absence of cortical binocularity in the hemisphere receiving direct geniculate input was not disrupted, indicates the absence of callosal transfer from the deafferented to the intact hemisphere.

Animals↗

The functional development of input-output relationships in the rostral portion of the corpus callosum in the kitten.

Microstimulation of the rostral portion of the corpus callosum (CC) was carried out on 21 awake kittens ranging in age from 45 to 105 days to determine the age at which motor responses first appeared and that at which they assumed functional adult-like properties. Motor responses to microstimulation first appeared over an interval ranging from 78-86 days postnatally. As in adults, they consisted of discrete, well-localized contractions of shoulder, whisker, and eyelid muscles according to the stimulated sites. In the first days after their appearance, motor responses differed markedly from those in adults because: (a) they exhibited higher thresholds; (b) they did not faithfully follow pulse trains delivered at 10 s intervals; (c) they had variable and longer latencies. Thereafter, motor responses gradually became stable, faithfully followed suprathreshold stimulation delivered at 0.1/s frequency, and acquired lower thresholds and shorter latencies, until they exhibited adult-like properties at 93-100 days of age. Single-unit recordings were obtained from 138 fibres isolated in the same callosal region submitted to microstimulation in order to study the response properties of the callosal fibres to somatic stimuli in immature animals. On the basis of their reactivity to peripheral stimulation, fibres were classified into three main types: (1) unreactive units (58 fibres), which could not be driven by somatic stimuli. (2) Adult-like units (55 fibres), which were readily driven by somatic stimuli and were endowed with fixed and small receptive fields (RFs) indistinguishable from those of adults. (3) Immature units (25 fibres), which were unsteadily driven by somatic stimuli applied over large areas at the periphery. Neither the RFs nor the adequate stimuli could be reliably determined. This type of units was not found in the adult cat (Spidalieri et al. 1985). The proportion of unreactive units was the highest before the appearance of motor responses and gradually decreased, approaching the adult level after attaining adult-like motor responses. Conversely, the proportion of adult-like units was lowest before the appearance of motor responses and gradually increased, approaching the adult level after motor responses had acquired adult-like properties.

Action Potentials↗

Split chiasm developmentally induced in kittens: plasticity of interhemispheric transfer in visual cortex cells.

Visual callosal transfer during development was studied in order to reveal plasticity-related compensation for the absence of direct contralateral inputs. The optic chiasm was midsagittally sectioned in 6-8 weeks old kittens (OCK) and for comparison, in adult cats (OCA). Unit recording was made during adulthood in the border area between visual cortex areas 17 and 18, namely the callosal projection zone. The proportion of cells showing interhemispheric transfer in the OCK group, as indicated by the presence of visual input from the contralateral eye was 10.5%; in the OCA cats their proportion was 4.0%. Moreover, 2.3% of the cells showed a pure transfer of input from the contralateral eye in the OCK, although none was seen in the OCA cats. Thus, during the developmental period, a plasticity induced process, albeit limited, takes place in the enhancement of interhemispheric transfer of visual information.

Animals↗

Neuropsychological assessment of the transcallosal approach.

The long-term consequences of partial callosal lesions were examined in 7 neurosurgically treated patients. Detailed clinical and neuropsychological assessment of the interhemispheric transfer (multimodal sensory and motor tasks) as well as memory and attention tests were used. The results revealed some disconnection symptoms with minor clinical significance, which could not be attributed to particular sites of the corpus callosum, except the splenium. It is questionable whether the reported memory and attention impairments are caused by the callosal lesion or by extracallosal pathology. The results indicate that the transcallosal approach is a safe and feasible alternative in the management of pathological midline processes in the brain.

Adolescent↗

The nature of transcallosal responses in the parietal cortex of kittens.

The nature of transcallosal responses in the parietal cortex was studied in experiments with kittens age 2 to 18 days, immobilized with Diplacin. The recorded potentials were represented by a positive-negative complex that was often accompanied by a latent negative oscillation. It was established that the earlier positive-negative oscillation was callosal in nature, and the later, extracallosal. As the kittens developed, we noted that currents responsible for the appearance of early and late negative oscillations converged in time. The results of layer-by-layer analysis of transcallosal responses showed that as the kittens developed, discharge of the surface positive component shifted from layer V to layer III, and discharge of the surface-negative remained at the II-III layer level. The late negative component was recorded only to the level of layers III-IV, having a discharge in layers I-II. Thus, the interhemispheric interrelations in the parietal cortex of kittens are realized by both callosal and extracallosal systems, and an increase in the degree of their interaction can be observed as the kittens grow.

Age Factors↗

Disturbances of cerebrospinal fluid (CSF) circulation--neuropsychiatric symptoms and neuroradiological contribution.

The present study aimed at relating dementia, pseudo-neurasthenic and affective organic brain syndromes to underlying type of CSF flow disorder and to subsequent alteration of anatomy. T2*-weighted magnetic resonance imaging (MRI) in the midsagittal plane permitted an analysis of aqueductal CSF flow phenomena and hydrocephalus-induced elevation, thinning and dorsal impingement of the corpus callosum. Furthermore, the width of the third ventricle was measured on the transverse scout images. 72 patients with communicating hydrocephalus (increased aqueductal CSF pulsations) and 26 patients with aqueductal stenosis (absence of aqueductal flow phenomena) were compared with 22 controls. Dementia and affective disorders were distributed equally among both CSF flow subgroups whereas pseudo-neurasthenic syndromes were observed more frequently in non-communicating hydrocephalus (p < 0.03). Alzheimer-type and multiinfarct dementia syndromes were found more frequently in communicating hydrocephalus whereas non-classifiable dementia showed some predilection for non-communicating hydrocephalus. Callosal height, area and third ventricular width did not predict affective or pseudoneurasthenic disorder whereas third ventricular width (p < 0.01) and callosal area (p < 0.05) discriminated between demented and non-demented patients. Dorsal impingement of the corpus callosum by the falx was a non-specific finding.

Cerebral Aqueduct↗

Lesions of the corpus callosum in hydrocephalic patients with ventricular drainage--a CT-study.

Patients with ventricular drainage may develop lesions of the corpus callosum. In order to study frequency, pathogenesis and clinical relevance of callosal lesions, 301 routine CT investigations of 79 patients with ventricular drainage were reviewed. Hypodense lesions in the anterior part of the corpus callosum were observed in 7 patients with longstanding hydrocephalus of variable origin, in 2 cases the corpus callosum was slightly swollen. All 7 patients showed signs of forced ventricular drainage. The callosal alterations were clinically inapparent and potentially reversible. Knowledge of these lesions is important to avoid misinterpretation.

Adolescent↗

Cortical binocularity in convergent strabismus after section of the optic chiasm.

In convergent strabismus (esotropia) the spatial asynchrony of the two eye inputs unbalances the interocular interactions, leading to the functional advantage of the nondeviated eye and the inhibition of the esotropic eye. It may be argued that the strabismic suppression, if it is the effect of inhibitory interactions between the eyes, could be removed by interrupting the interocular pathways at the optic chiasm. After chiasmatic section, each eye is connected only to the ipsilateral cortex through the uncrossed retinal projections and so the functionality of each eye's input is no longer interfered with by interocular mechanisms. In strabismic cats submitted, as adults, to section of the optic chiasm, we performed electrophysiological recordings from the striate cortex. Results show that in these animals: (1) the cortical responsiveness to the strabismic eye is strikingly higher than in esotropes with an intact optic chiasm; (2) the effectiveness of stimulation of the deviated eye is not different from that of the nondeviated eye in driving neurons of corresponding cortex; (3) surprisingly, a high degree of binocular activation is present in the cortex ipsilateral to the deviated eye, while in the cortex connected to the nondeviated eye the greatest majority of neurons are monocularly driven. Cortical binocularity depends on the corpus callosum, which conveys the input from the nondeviated eye to the opposite cortex (which receives the direct strabismic input), but not vice versa. The asymmetry of callosal transmission parallels the morphological asymmetry of callosal connections that occur in convergent strabismus. All together the findings indicate that the impaired effectiveness of esotropic input does not result from developmental deficit of the strabismic afferents but, rather, from inhibitory influences that are actively exerted through the interocular pathways. Strabismic suppression may be accomplished by the same interocular mechanisms underlying binocular rivalry.

Animals↗

Pathways of interhemispheric transfer in normals and in a split-brain subject. A positron emission tomography study.

We studied with PET the intra- and interhemispheric pathways subserving a simple, speeded-up visuomotor task. Six normal subjects and one patient with a complete section of the corpus callosum (M.E.) underwent regional cerebral blood flow (rCBF) measurements under conditions of lateralized tachistoscopic visual presentations in a simple manual reaction time paradigm. Confirming previous behavioural findings, we found that on average crossed hand and/or hemifield conditions, i.e. those requiring an interhemispheric transfer of information, yielded a longer RT than uncrossed conditions. This difference (0.7 ms) was dramatically larger (45.6 ms) in the callosum-sectioned patient M.E. In normal subjects the cortical areas selectively activated in uncrossed and crossed conditions were different. In the former condition, most activation foci were anterior to the ventral anterior commissure (VAC) plane, whereas in the latter there was a prevalent parietal and occipital activation. This shows that a simple model in which the cortical visuo-motor pathways are similar in the intra- and the interhemispheric condition, with an extra callosal route for the latter, is too simplistic. Furthermore, these results suggest that the bulk of visuomotor interhemispheric transfer takes place through the widespread callosal fibres interconnecting the parietal cortices of the two hemispheres. The pattern of activation in the two crossing conditions was markedly different in M.E., in whom interhemispheric transfer might take place via his intact anterior commissure or subcortical commissures.

Adult↗

Interobserver agreement for diagnostic MRI criteria in suspected multiple sclerosis.

MRI is the paraclinical test most widely used to support the diagnosis of multiple sclerosis (MS). We evaluated interobserver agreement in applying diagnostic criteria to MRI obtained at first presentation. Five experienced observers scored 25 sets of images consisting of unenhanced T2- and gadolinium-enhanced T1-weighted images (approximately half the sets were normal). We scored frontal, parietal, temporal, occipital, infratentorial and basal ganglia lesions and the total number of lesions on T2-weighted images; periventricular, callosal, juxtacortical and ovoid lesions and those > 5 mm in maximum diameter; contrast-enhancing and hypointense lesions. Based on a combination of imaging findings patients were classified as compatible or not compatible with MS according to composite criteria. Observer concordance was characterised by weighted kappa values (kappa) and mean average difference to the median (MADM) scores. Using the raw scores, there was poor agreement for the total number of lesions on T2-weighted images, and for occipital, oval, juxtacortical and hypointense lesions. Moderate agreement was found for frontal, callosal, basal ganglia and large lesions on T2 weighting. Good agreement was attained for parietal, temporal, infratentorial and periventricular lesions. After dichotomisation according to accepted cut-off values, most criteria performed better, especially the number of lesions on T2-weighted images (P < 0.05). Good agreement was found for the criteria of Paty and Fazekas and moderate agreement for those of Barkhof. While experienced observers may not agree on the total number of lesions, they show quite good agreement for commonly used cut-off points and elements in the composite criteria. This validates the use of MRI in the diagnosis of MS, and the use of dichotomised and composite criteria.

Brain↗

Cell-wall antigens in mesophyll cells and mesophyll-derived protoplasts of sugar beet: possible implication in protoplast recalcitrance?

We have investigated the possible relation between plant cell-wall constituents and the recalcitrance of the cell to regenerate organs and whole plants in vitro. A temporal and spatial expression of several carbohydrate epitopes was observed both within leaf tissue used for protoplast isolation and within new walls reformed by recalcitrant mesophyll protoplasts of sugar beet ( Beta vulgaris L.); these include four pectic epitopes, one xyloglucan (rhamnogalacturonan I) epitope, two carbohydrate motifs of arabinogalactan proteins (AGPs) and callose. The walls of mesophyll cells and newly formed walls of protoplasts were similar with respect to the presence of large amounts of pectins recognized by JIM7 antibodies, the scarcity of JIM5-pectins and the complete absence of LM5-responding pectin molecules. Their main differences were the significantly higher accumulation of LM6-recognizing pectins and the very conspicuous greater accumulation of AGPs and callose in walls deposited by protoplasts than in those synthesized by donor cells.

Antigens↗

Clinicoradiologic subtypes of Marchiafava-Bignami disease.

The clinical diagnosis of Marchiafava-Bignami disease (MBD) has considerably changed during recent decades with brain MRI providing the opportunity of a reliable in-vivo diagnosis. However, semiologic and neuroimaging characteristics of the currently known spectrum of MBD have not been investigated systematically, and knowledge of clinicoradiologic associations is sketchy. We report an illustrative case with limited callosal involvement on MRI and a favorable outcome and have reviewed literature on clinical and radiologic features in 50 cases of MBD diagnosed in vivo since 1985. Our reviewed data suggest the differentiation of two clinicoradiologic subtypes: Type A is characterized by major impairment of consciousness, T2-hyperintense swelling of the entire corpus callosum on early MRI and poor outcome. Type B shows at most slight impairment of consciousness, partial callosal lesions on MRI and a favorable outcome. Differentiation of these clinicoradiologic subtypes may help resolve inconsistencies of the established clinical classification resulting from new insights into the clinical course and prognosis of MBD by structural neuroimaging.

Adult↗

A putative plant homolog of the yeast beta-1,3-glucan synthase subunit FKS1 from cotton (Gossypium hirsutum L.) fibers.

A novel plant gene CFL1 was cloned from cotton (Gossypium hirsutum L.) fibers by expressed sequence tag (EST) database searching and 5'-RACE (rapid amplification of cDNA ends). This gene shows sequence homology with FKS1 which has been identified as the putative catalytic subunit of the yeast beta-1,3-glucan synthase. It encodes a protein (CFL1p) of 219 kDa with 13 deduced transmembrane helices and 2 large hydrophilic domains, one of which is at the N-terminus and the other in the internal region of the polypeptide. CFL1 displays 21% identity and 41% similarity to FKS1 at the amino acid level over its entire length, with 31% identity and 52% similarity for the hydrophilic central domain. Using RNA and protein blot analysis, CFL1 was found to be expressed at higher levels in cotton fibers during primary wall development. CFL1 also had a strong expression in young roots. Using a calmodulin (CaM)-gel overlay assay, the hydrophilic N-terminal domain of CFL1p was shown to bind to CaM, while the hydrophilic central domain did not. A putative CaM-binding domain, 16 amino acids long, was predicted in the hydrophilic N-terminal domain. Moreover, a product-entrapment assay demonstrated that a protein associated with an in vitro-synthesized callose pellet could be labeled by anti-CFL1 antibodies. Our finding suggests that CFL1 is a putative plant homolog of the yeast beta-1,3-glucan synthase subunit FKS1 and could be involved in callose synthesis.

Calmodulin-Binding Proteins↗