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Interactions among neuronal systems assessed with functional neuroimaging.

Cortical organisation is based on the principles of functional specialisation and functional integration. Those two concepts exist at multiple spatial scales. For example at a macroscopic level, functionally segregated areas V1 and V5 are integrated within the dorsal visual stream. At a microscopic level, it is possible to infer the functional integration of (segregated) ocular dominance columns within area V1. At the macroscopic level functional specialisation and integration can be tested with functional neuroimaging, using fMRI/PET or EEG/MEG. A framework that allows making inferences on functional integration between brain regions using fMRI will be presented. The common feature of all techniques within that framework is that they can incorporate second order terms and therefore explicitly allow for contextual modulations. Two examples demonstrate how attention and paired associates learning can modulate effective connectivity within the visual system. Furthermore these models can incorporate non-linear modulatory (feedback) connections that can account for the changes in effective connectivity observed.

Attention↗

Transesophageal echocardiographic assessment of left ventricular function during apnea testing for brain death.

The effects of apnea testing-induced respiratory acidosis on left ventricular function (LVF) are still controversial. The aim of the study was to assess LVF during apnea testing using transesophageal echocardiography (TEE). Twenty consecutive patients suspected of brain death, hemodynamically stable, and considered as potential organ donors were prospectively studied. A 20-min apnea test was performed after obtaining a PaCO2 > 35 mmHg and 20 min of FIO2 1 ventilation. LVF was assessed using TEE with a CFM 750 (Diasonic) connected to a 5 MHz probe. Heart rate (HR), mean arterial pressure (MAP), left ventricle end-diastolic and systolic area (LVEDA, LVESA), and LVF assessed by fractional area changes (FAC), systolic wall motion (SWM) scores, and blood gases were recorded at baseline, and after 5, 10, 15, and 20 min of apnea testing. In 19 patients, no spontaneous respiratory movement occurred during the standard 20-min period. In one patient (No. 15), the apnea test had to be stopped after 10 min because of hypoxia. HR, LVEDA, LVESA, and SWM were not significantly modified during the study. There was a progressive statistically significant decrease in MAP during apnea (from 77 +/- 10 to 63 +/- 11 mmHg), associated with a statistically significant increase in FAC at 20 min (from 48 +/- 13 to 56 +/- 8%). PaCO2 progressively rose (from 40 +/- 3 to 95 +/- 11 mmHg), associated with a decrease in pH (from 7.42 +/- 0.06 to 7.09 +/- 0.08). At the same time, PaO2 decreased slightly in all patients, but values remained well above hypoxic levels, except for one patient. Despite severe respiratory acidosis the increase in FAC suggests that apnea testing is well tolerated for brain death assessment.

Acidosis, Respiratory↗

[Organization of cortico-reticulo-spinal connections in the cat].

Time characteristics of the monosynaptic EPSP evoked by stimulation of the motor cortex in reticulospinal neurons were studied in detail. Negative correlation was revealed between latency duration of EPSP and conduction velocity of the corticobulbar fibres and positive correlation between such characteristics and conduction velocity of axons. Reticulospinal neurons with conduction velocity from 10.8 to 65.0 m/s were activated by fast and slow corticobulbar fibres, but neurons with conduction velocity from 65.0 to 155 m/s were activated only by slow ones. A functional role of different cortico-reticulo-spinal connections is discussed.

Animals↗

Expression and mapping of lubricin in canine flexor tendon.

Matrix composition and the biomechanical environment are intimately interdependent in most connective tissues. Lubricin has many distinct biological functions, including lubrication, antiadhesion, and cytoprotection in cartilage, tendons, and other tissues. This study investigated the distribution of lubricin in the canine flexor digitorum profundus (FDP) tendon by immunohistochemistry. Lubricin was found both on the tendon surface and at the interface of collagen fiber bundles within the tendon, where the cells are subjected to shear force in addition to tension and compression. The expression of lubricin in regions of the canine flexor tendon that differ in mechanical or nutritional environment was also investigated using RT-PCR. Six N-terminal splicing variants were identified from six distinct anatomical regions of flexor tendon. The variants with larger sizes were noted in regions subjected to significant shear and compressive forces. Lubricin is ubiquitous in the FDP tendon, with variations in distribution and splicing that appear to correspond to discrete anatomic locations that differ by mechanical or nutritional environment.

Animals↗

Comparative linkage maps of the rice and maize genomes.

Genetic linkage maps have been constructed for the rice and maize genomes on the basis of orthologous loci detected with a common set of cDNA clones. Conserved linkage groups could be identified, which together account for more than two-thirds of both genomes. In some instances, entire chromosomes or chromosome arms are nearly identical with respect to gene order and gene content. The results also reveal that most of the genes (> 72%) duplicated during ancient polyploidization are still present in the maize genome in duplicate copy. The comparative maps of rice and maize provide a basis for interpreting molecular, genetic, and breeding information between these two important species and establish a framework for ultimately connecting the genetics of all grass species.

Biological Evolution↗

The dissonance mutant of courtship song in Drosophila melanogaster: isolation, behavior and cytogenetics.

The dissonance mutant of courtship song was induced by chemical mutagenesis. This X-chromosomal mutation causes the D. melanogaster male's acoustical output, resulting from his wing vibrations directed at a female, to include very long and loud tone "pulses." Yet, a given train of pulses starts out as normal, with the signals in all but the shortest singing bouts eventually becoming polycyclic and high-amplitude. The aberrant songs caused by diss (map position, 1-52; cytological interval, 14C1-2 to 14C4-5) were quantitatively compared to those produced by mutant cacophony males, whose pulses are much more uniformly polycyclic (due to a mutation mapping elsewhere on the X chromosome). Males or females expressing diss are normal in several "general" behaviors. Yet diss males not only sing abnormally, but they also exhibit longer-than-normal mating latencies in their courtship of females. These decrements seem to be associated, at least in part, with visually aberrant behavior of diss flies--measured with regard to male courtship per se, and also in tests of more general visual responses. Such defects were found when testing diss males or females, and the genetic etiology of the visual impairments were provisionally mapped to the same locus to which the song abnormality has been localized. Neurogenetic connections between the control of courtship singing behavior and visual system functions are discussed with respect to the new song mutation (diss) and the older one (cac)--which also turned out to be genetically related to a mutation that causes abnormalities of light-induced behavior and physiology.

Animals↗

Functional Properties of Neurons in the Anterior Bank of the Parieto-occipital Sulcus of the Macaque Monkey.

Extracellular recordings were made in the anterior bank of the parieto-occipital sulcus of two waking monkeys trained to perform fixation tasks in normal illumination or in complete darkness. Of the recorded neurons, 73% (251/343) were responsive to visual stimulation, but their overall organization did not conform to a simple, continuous retinotopic map. Most of the visual neurons showed a high degree of orientation and direction sensitivity, higher than that found in areas V1, V2 and V3A under the same experimental conditions. Whether they had a resolvable receptive field or not, the discharge rate of many neurons in the anterior bank of the parieto-occipital sulcus was influenced by oculomotor activity. The animals were required to execute pursuit or saccadic eye movements in darkness. Saccadic eye movements were found to influence 19% of the neurons tested (29/156); by contrast, pursuit eye movements were without effect (0/64). Saccade responses were direction-tuned and, in several cases, the neuronal discharge started before the onset of eye movement. The animals were also required to gaze, in darkness, at nine different positions on the screen they faced. Of the neurons tested, 59% (102/174) were affected by the direction of gaze. Higher discharge rates were generally observed when the animals looked towards the lower part of the field of view. Given the functional properties of its neurons, its connections with area V3A-where neural signals appropriate for building an objective map of the visual space are present (Galletti and Battaglini, 1989, J. Neurosci., 9, 1112-1125)-and its output to the visuomotor centres involved in the generation of saccades (frontal eye fields and superior colliculus), we infer that the cortex of the anterior bank of the parieto-occipital sulcus might be part of the network involved in the control of gaze in order to locate objects in visual space.

Journal Article↗

The development of the nucleus isthmi in Xenopus laevis. I. Cell genesis and the formation of connections with the tectum.

The nucleus isthmi (NI) of the amphibian relays visual input from one tectum to the other tectum and thus brings a visual map from the eye to the ipsilateral tectum. This isthmotectal visual map develops slowly; it is first detected electrophysiologically at stages 60-62, the age at which the eyes begin their dorsalward migration and the region of binocular overlap beings to increase in extent. During this critical period of life, normal binocular visual input is required for establishment of normal topographic isthmotectal projections. In this study, we have used anatomical methods to trace cell birth, cell death, and formation of connections by the nucleus isthmi during the critical period. Tritiated thymidine labelling demonstrates that cells in the nucleus isthmi are generated throughout most of tadpole life (stages 29-62). Most cells conform to an orderly ventrodorsal gradient starting from stage 29 and extending to stages 56; later cells are inserted at apparently random locations in the nucleus. We have re-examined the hypothesis of Tay and Straznicky ('80) that the order of cell genesis in the NI and tectum could help establish proper isthmotectal connections, and we find that a timing mechanisms does not explain the two-dimensional topography of the isthmotectal map but that timing may aid in proper mediolateral positioning of isthmotectal axons at the points where they first enter the tectum. Horseradish peroxidase labelling was used to investigate whether anatomical projections from tectum to NI and from NI to tectum are present prior to the onset of eye migration. The results show that there are tectoisthmotectal projections by stage 52. Moreover, isthmotectal axons grow into as yet monocular tectal regions prior to the onset of eye migration. At stage 60, when binocular overlap begins, isthmotectal axons are visible throughout the tectum but are densely branched only at the rostral tectal margin, the location where they are predicted to occur on the basis of electrophysiological maps.

Acetylcholinesterase↗

A three-dimensional multivariate image processing technique for the analysis of FTIR spectroscopic images of multiple tissue sections.

BACKGROUND: Three-dimensional (3D) multivariate Fourier Transform Infrared (FTIR) image maps of tissue sections are presented. A villoglandular adenocarcinoma from a cervical biopsy with a number of interesting anatomical features was used as a model system to demonstrate the efficacy of the technique. METHODS: Four FTIR images recorded using a focal plane array detector of adjacent tissue sections were stitched together using a MATLAB routine and placed in a single data matrix for multivariate analysis using Cytospec. Unsupervised Hierarchical Cluster Analysis (UHCA) was performed simultaneously on all 4 sections and 4 clusters plotted. The four UHCA maps were then stacked together and interpolated with a box function using SCIRun software. RESULTS: The resultant 3D-images can be rotated in three-dimensions, sliced and made semi-transparent to view the internal structure of the tissue block. A number of anatomical and histopathological features including connective tissue, red blood cells, inflammatory exudate and glandular cells could be identified in the cluster maps and correlated with Hematoxylin & Eosin stained sections. The mean extracted spectra from individual clusters provide macromolecular information on tissue components. CONCLUSION: 3D-multivariate imaging provides a new avenue to study the shape and penetration of important anatomical and histopathological features based on the underlying macromolecular chemistry and therefore has clear potential in biology and medicine.

Journal Article↗

Development of the atrioventricular valve tension apparatus in the human heart.

Using various microscopical techniques we studied the development of the atrioventricular valves in human hearts between 5 and 19 weeks of development. Within the atrioventricular cushions two different layers could be recognized that remained present in all ages studied. The atrial layer, being present at the side of the atrioventricular orifice, was positive for laminin while the ventricular layer, that was connected to the myocardium, was positive for fibronectin and collagen III. Fate-mapping of these two layers, morphometrics, and scanning electron microscopy, supplemented with in vivo labeling of cushion tissue in chicken hearts have lead to new insights in the process of valve development. The cushions became freely movable prevalvular leaflets by delamination of ventricular myocardium underneath the cushion tissue. This myocardium gradually retracted towards annulus and papillary muscles and finally disappeared, resulting in fibrous, non-myocardial valves. The atrial layer of the cushions remained present as a jelly-like surface on the valve leaflets while the ventricular layer of the cushions became the compact fibrous tissue of the leaflets and the chords. Chordal development was first visible at 10 weeks of development when gaps were formed in the ventricular layer of the cushions on top of the papillary muscles. These gaps enlarged into the interchordal spaces while the cushion tissue in between the gaps lengthened to form the chords. We conclude that the leaflets as well as the chords of the atrioventricular valves are derived from atrioventricular cushion tissue. Myocardium is only important for loosening of the leaflets while keeping connection with the developing papillary muscles. Errors in delamination or retraction of myocardium or remodeling of cushion tissue into chords form the basis for various congenital valve anomalies.

Embryonic and Fetal Development↗

Loss of polyneuronal innervation and establishment of a topographical map in the glutaeus muscle of Bufo marinus during generation of secondary muscle cells.

The development of synaptic connections to the toad (Bufo marinus) glutaeus magnus from segmental nerves 8 (N8) and 9 (N9) was determined in the postmetamorphic period. Three different-size toads were studied: small (0.3-2.0 g), medium-size (5-15 g) and large (greater than 20 g). The number of cells in the glutaeus increased about 9-fold during development; this involved the appearance and subsequent maturation of secondary fibres throughout the muscle. The glutaeus in small toads, which consisted almost entirely of primary fibres, was innervated to a similar extent by N8 and N9 as assessed by tetanic contraction measurements. During late development there was a progressive increase in the percentage of the muscle innervated by N9 and a decrease in the percentage innervated by N8. This change in the segmental innervation was accompanied by changes in the innervation of the ventral glutaeus as assessed by intracellular recording. In small toads this surface of the muscle was innervated predominantly by N8, with N9 frequently appearing as a low-efficacy terminal on dually innervated fibres. With further development there was a progressive reduction in the percentage of dually innervated fibres and a concomitant decrease in the percentage innervation of the entire ventral glutaeus by N8. These results suggest that the topographical projection is established by the initial distribution of N9 terminals on the primary fibres of the muscle. The multiple innervation of newly generated fibres and the on-going process of terminal elimination results in N9 terminals, many of which were initially weak, preserving their position in the muscle. This occurs at the expense of N8 terminals, whose relative incidence declines during development. The competitive advantage of N9 motoneurones may be due to their greater capacity to lay down axon collaterals and preferentially innervate newly generated fibres; alternatively N9 terminals may displace N8 terminals, which were initially more efficacious, from dually innervated fibres. Secondary muscle fibres generated throughout the muscle are thus incorporated into an increasingly precise topographical map.

Animals↗

Synaptic potentials evoked by convergent somatosensory and corticocortical inputs in raccoon somatosensory cortex: substrates for plasticity.

1. "Unmasking" of weak synaptic connections has been suggested as a mechanism for the early changes in cortical topographic maps that follow alterations of sensory activity. For such a mechanism to operate, convergent sensory inputs must already exist in the normal cortex. 2. We tested for topographic and cross-modality convergence in primary somatosensory cortex of raccoon. The representation of glabrous skin of forepaw digits was chosen because, even though it is dominated by inputs from the glabrous skin of a single digit, it nevertheless comes to respond to stimulation of other digits when, e.g., a digit is removed. 3. Intracellular recordings were made from 109 neurons in the representation of glabrous skin of digit 4. Neurons were tested for somatosensory inputs with electrical and natural stimulation of digits. 4. Excitatory postsynaptic potentials (EPSPs) were evoked in 100% of the neurons (109/109) by electrical stimulation of glabrous skin of digit 4, and in 79% (31 of 39) by vibrotactile stimulation. 5. Glabrous skin of digit 4 was not the sole source of somatosensory inputs. A minority of neurons generated EPSPs after electrical stimulation of hairy skin of digit 4 (10 of 98 neurons, 10%). Electrical stimulation of digits 3 or 5 evoked EPSPs in 22 of 103 neurons (21%). Natural stimulation (vibrotactile or hair bending) was also effective in most of these latter cases (digit 3, 6/7; digit 5, 9/10). 6. Intracortical microstimulation of the "heterogeneous zone" was used to test for corticocortical connections to neurons in the glabrous zone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Functional MRI in human motor control studies and clinical applications.

Functional magnetic resonance imaging (fMRI) has been a useful tool for the noninvasive mapping of brain function associated with various motor and cognitive tasks. Because fMRI is based on the blood oxygenation level dependent (BOLD) effect, it does not directly record neural activity. With the fMRI technique, distinguishing BOLD signals created by cortical projection neurons from those created by intracortical neurons appears to be difficult. Two major experimental designs are used in fMRI studies: block designs and event-related designs. Block-designed fMRI presupposes the steady state of regional cerebral blood flow and has been applied to examinations of brain activation caused by tasks requiring sustained or repetitive movements. By contrast, the more recently developed event-related fMRI with time resolution of a few seconds allows the mapping of brain activation associated with a single movement according to the transient aspects of the hemodynamic response. Increasing evidence suggests that multiple motor areas are engaged in a networked manner to execute various motor acts. In order to understand functional brain maps, it is important that one understands sequential and parallel organizations of anatomical connections between multiple motor areas. In fMRI studies of complex motor tasks, elementary parameters such as movement length, force, velocity, acceleration and frequency should be controlled, because inconsistency in those parameters may alter the extent and intensity of motor cortical activation, confounding interpretation of the findings obtained. In addition to initiation of movements, termination of movements plays an important role in the successful achievement of complex movements. Brain areas exclusively related to the termination of movements have been, for the first time, uncovered with an event-related fMRI technique. We propose the application of fMRI to the elucidation of the pathophysiology of movement disorders, particularly dystonia, which exhibits involuntary co-contraction of agonist and antagonist muscles and manifests abnormal posture or slow repetition of movements.

Brain Mapping↗

A genetic screen for synaptic transmission mutants mapping to the right arm of chromosome 3 in Drosophila.

Neuronal function depends upon the proper formation of synaptic connections and rapid communication at these sites, primarily through the regulated exocytosis of chemical neurotransmitters. Recent biochemical and genomic studies have identified a large number of candidate molecules that may function in these processes. To complement these studies, we are pursuing a genetic approach to identify genes affecting synaptic transmission in the Drosophila visual system. Our screening approach involves a recently described genetic method allowing efficient production of mosaic flies whose eyes are entirely homozygous for a mutagenized chromosome arm. From a screen of 42,500 mutagenized flies, 32 mutations on chromosome 3R that confer synaptic transmission defects in the visual system were recovered. These mutations represent 14 complementation groups, of which at least 9 also appear to perform functional roles outside of the eye. Three of these complementation groups disrupt photoreceptor axonal projection, whereas the remaining complementation groups confer presynaptic defects in synaptic transmission without detectably altering photoreceptor structure. Mapping and complementation testing with candidate mutations revealed new alleles of the neuronal fate determinant svp and the synaptic vesicle trafficking component lap among the collection of mutants recovered in this screen. Given the tools available for investigation of synaptic function in Drosophila, these mutants represent a valuable resource for future analysis of synapse development and function.

Animals↗

Natural variation for carbohydrate content in Arabidopsis. Interaction with complex traits dissected by quantitative genetics.

Besides being a metabolic fuel, carbohydrates play important roles in plant growth and development, in stress responses, and as signal molecules. We exploited natural variation in Arabidopsis (Arabidopsis thaliana) to decipher the genetic architecture determining carbohydrate content. A quantitative trait locus (QTL) approach in the Bay-0 x Shahdara progeny grown in two contrasting nitrogen environments led to the identification of 39 QTLs for starch, glucose, fructose, and sucrose contents representing at least 14 distinct polymorphic loci. A major QTL for fructose content (FR3.4) and a QTL for starch content (ST3.4) were confirmed in heterogeneous inbred families. Several genes associated with carbon (C) metabolism colocalize with the identified QTL. QTLs for senescence-related traits, and for flowering time, water status, and nitrogen-related traits, previously detected with the same genetic material, colocalize with C-related QTLs. These colocalizations reflect the complex interactions of C metabolism with other physiological processes. QTL fine-mapping and cloning could thus lead soon to the identification of genes potentially involved in the control of different connected physiological processes.

Arabidopsis↗

Fusing EEG and fMRI based on a bottom-up model: inferring activation and effective connectivity in neural masses.

The elucidation of the complex machinery used by the human brain to segregate and integrate information while performing high cognitive functions is a subject of imminent future consequences. The most significant contributions to date in this field, known as cognitive neuroscience, have been achieved by using innovative neuroimaging techniques, such as electroencephalogram (EEG) and functional magnetic resonance imaging (fMRI), which measure variations in both the time and the space of some interpretable physical magnitudes. Extraordinary maps of cerebral activation involving function-restricted brain areas, as well as graphs of the functional connectivity between them, have been obtained from EEG and fMRI data by solving some spatio-temporal inverse problems, which constitutes a top-down approach. However, in many cases, a natural bridge between these maps/graphs and the causal physiological processes is lacking, leading to some misunderstandings in their interpretation. Recent advances in the comprehension of the underlying physiological mechanisms associated with different cerebral scales have provided researchers with an excellent scenario to develop sophisticated biophysical models that permit an integration of these neuroimage modalities, which must share a common aetiology. This paper proposes a bottom-up approach, involving physiological parameters in a specific mesoscopic dynamic equations system. Further observation equations encapsulating the relationship between the mesostates and the EEG/fMRI data are obtained on the basis of the physical foundations of these techniques. A methodology for the estimation of parameters from fused EEG/fMRI data is also presented. In this context, the concepts of activation and effective connectivity are carefully revised. This new approach permits us to examine and discuss some future prospects for the integration of multimodal neuroimages.

Brain↗