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Double-stranded RNA induces mRNA degradation in Trypanosoma brucei.

Double-stranded RNA (dsRNA) recently has been shown to give rise to genetic interference in Caenorhabditis elegans and also is likely to be the basis for phenotypic cosuppression in plants in certain instances. While constructing a plasmid vector for transfection of trypanosome cells, we serendipitously discovered that in vivo expression of dsRNA of the alpha-tubulin mRNA 5' untranslated region (5' UTR) led to multinucleated cells with striking morphological alterations and a specific block of cytokinesis. Transfection of synthetic alpha-tubulin 5' UTR dsRNA, but not of either strand individually, caused the same phenotype. On dsRNA transfection, tubulin mRNA, but not the corresponding pre-mRNA, was rapidly and specifically degraded, leading to a deficit of alpha-tubulin synthesis. The transfected cells were no longer capable of carrying out cytokinesis and eventually died. Analysis of cytoskeletal structures from these trypanosomes revealed defects in the microtubules of the flagellar axoneme and of the flagellar attachment zone, a complex cortical structure that we propose is essential for establishing the path of the cleavage furrow at cytokinesis. Last, dsRNA-mediated mRNA degradation is not restricted to alpha-tubulin mRNA but can be applied to other cellular mRNAs, thus establishing a powerful tool to genetically manipulate these important protozoan parasites.

Animals↗

Altered corticogenesis and neuronal morphology in irradiation-induced cortical dysplasia: a Golgi-Cox study.

Cortical dysplasia has a strong clinical association with epilepsy and mental retardation, but the relationship between alterations in cortical structure and function in dysplasia-related disorders is poorly understood. The cerebral cortex of irradiated rats, an experimental model of cortical dysplasia, was studied using cresyl violet-stained sections and the Golgi-Cox method. The irradiated cortex is characterized by reductions in size, volume, and number of neurons and fibers reflecting the original lethal injury to neuronal precursors. Consequently, only neurons that survived this injury were able to continue their, albeit altered, development. The result is an altered corticogenesis characterized by neuronal, fiber circuitry, and microvascular alterations. Abnormal aggregates (nodules) of excitatory pyramidal neurons with altered dendritic profiles and functional territories are found between 200 and 400 microm from the pial surface. Their horizontal dendritic profiles and functional territories contrast with the vertical (columnar) dendritic profiles and functional territories of normal pyramidal neurons. This horizontal concentration of spiny dendrites and, hence, of excitatory synaptic contacts suggests a response to the presence of an abnormal horizontal plexus of afferent fibers terminals. Stellate neurons, some morphologically compatible with inhibitory basket cells, are also essential components of these nodules. Some neuronal nodules are characterized by a rich plexus of anastomotic capillaries that contrasts with the sparser vasculature of surrounding gray matter tissue. The presence of well-vascularized aggregates of altered pyramidal and inhibitory neurons suggests a high level of metabolic activity. Well-vascularized deep heterotopias are also found. We propose that the functional activity of well-vascularized neuronal nodules and heterotopias could play a role in the abnormal cortical function in this model.

Aging↗

Absence of bundle structure in the neocortex of the reeler mouse at the embryonic stage. Studies by scanning electron microscopic fractography.

The reeler mutant mouse is characterized by a derangement of the cerebral cortical structure due to abnormalities during the migration step at the embryonic stage. We have analyzed both the control and reeler cerebral cortex by means of scanning electron microscopic fractography. In the control cerebral cortex, the bundle formation was composed of fine fibers on which the migrating neuroblasts were attached perpendicular to the pial surface, whereas no bundle formation was observed in the reeler; instead, there was a fine meshwork of fibers surrounding the neuroblasts. The possible role of bundle formation in the normal cerebral cortex and the correlation between the inability of cells to migrate and the absence of bundle formation in the reeler is discussed.

Animals↗

Functional evaluation of arteriovenous malformations.

Detailed knowledge of the angioarchitecture of arteriovenous malformations (AVMs) is necessary in determining the optimal timing and method of treatment of these challenging lesions. Many techniques are available for studying the functionality of surrounding cortical structures of AVMs. These include the use of positron emission tomography, functional magnetic resonance imaging, magnetoencephalography, and direct provocative testing of cortical function. The use of these methods to determine flow dynamics and tissue perfusion is also reviewed. These techniques are discussed in the present study, and their judicious utilization will enhance the safety of AVM therapy.

Amobarbital↗

Structural (operational) synchrony of EEG alpha activity during an auditory memory task.

Memory paradigms are often used in psycho-physiological experiments in order to understand the neural basis underlying cognitive processes. One of the fundamental problems encountered in memory research is how specific and complementary cortical structures interact with each other during episodic encoding and retrieval. A key aspect of the research described below was estimating the coupling of rapid transition processes (in terms of EEG description) which occur in separate cortical areas rather than estimating the routine phase-frequency synchrony in terms of correlation and coherency. It is assumed that these rapid transition processes in the EEG amplitude correspond to the "switching on/off" of brain elemental operations. By making a quantitative estimate of the EEG structural synchrony of alpha-band power between different EEG channels, it was shown that short-term memory has the emergent property of a multiregional neuronal network, and is not the product of strictly hierarchical processing based on convergence through association regions. Moreover, it was demonstrated that the dynamic temporal structure of alpha activity is strongly correlated to the dynamic structure of working memory.

Acoustic Stimulation↗

[Electrophysiologic study of thalamo-cortical connections during early postnatal ontogeny in cats].

Evoke potentials (EP) in the sensomotor, parietal, temporal and occipital areas of the cortex induced by the stimulation of thalamic nuclei (LP, VL, CL) were widely spread over the brain in kittens (6 to 41--day old) under nembutal anesthesia. Decreases of the EP latency and duration occurred mainly in those cortical structures into which direct protections of the thalamic were formed during ontogenesis. No significant changes in the EP characteristics occurred in other cortical zones. Dynamics of the EP characteristics in the cortex elicited by stimulating of the thalamic nuclei seems to reflect functional maturation of thalamocortical connections during early postnatal ontogenesis.

Animals↗

Neural topography and chronology of memory consolidation: a review of functional inactivation findings.

Findings on the role of subcortical and cortical structures in mnemonic processes, obtained by means of the reversible functional inactivation technique, are reviewed. The main advantage of this method (subcortical or cortical administration of local anesthetics or tetrodotoxin) is that it provides information not only on "where" but also "when" and for "how long" these processes take place, thus adding to the topographical dimension the chronological one. The review covers several types of memory (e.g., passive avoidance and spatial memory) studies examining the neural substrates of memory consolidation on the basis of the functional inactivation of the nucleus of the solitary tract, parabrachial nuclei, substantia nigra, hippocampus (dorsal and ventral), nucleus basalis magnocellularis, amygdala, medial septal area, striatum, olfactory bulb, and neocortex. The data are discussed in relation to earlier research and with respect to the anatomical and functional connectivity of the examined centers.

Anesthetics, Local↗

Interictal afterdischarge in focal penicillin epilepsy: thalamocortical unit activity.

The involvement of thalamic versus cortical structures for the initiation and maintenance of brief interictal afterdischarge was evaluated by recording extracellularly units and field potentials from different subcortical and cortical sites. Afterdischarge oscillations at 16 to 22/s that followed interictal spikes with a delay of 170 to 220 ms usually appeared 10 to 30 min after the topical application of penicillin to the cat's precruciate cortex. Units in the ventrolateral nucleus of the thalamus fired in burst discharges during cortical afterdischarge and less reliably during the cortical interictal spike. In contrast, units recorded at the cortical penicillin focus and homologous contralateral site remained silent during afterdischarge but had a typical burst discharge during the interictal spike. Although these data support a thalamic basis for the rhythm, the lack of an afterdischarge-like oscillation in the thalamic field potential and the independent appearance of afterdischarge and cortical recruiting waves elicited by stimulation of the nucleus reticularis would favor its cortical origin. In accordance with its frequency characteristics and data gained from earlier cooling studies we suggest a cortical mechanism requiring thalamic triggering for the generation of afterdischarge.

Animals↗

[Parameters of voluntary effort correlates with functional state of the motor control system].

The spectrum oscillations of a prolonged supported effort that was registered at isometric regimen in 3 age groups of healthy volunteers, were analyzed. Changes in distribution of spectral density of effort oscillations and distinction in distribution of spectral density as a reaction to fatigue in age groups, were shown. The revealed amplitude-frequency ranges of changes of spectral density of effort oscillations characterize activity of suprasegmental and segmental levels of motor control system fulfilling the voluntary movement control and the automatic regulation of posture. The correlates with functional state of motor control system are considered in the terms of voluntary and involuntary components of regulation. The substantial growth of activity in the central structures of motor control system in process of development of fatigue and the narrowing of the frequency range of activity of sub-cortical structures with age, were revealed.

Adolescent↗

Effects of cooling rate and glycerol concentration on the structure of the frozen kidney: assessment by cryo-scanning electron microscopy.

An experimental technique, employing a directional solidification stage for controlled freezing of tissue samples and low-temperature scanning electron microscopy for observation of the structure of the frozen-hydrated samples, was used to study freezing processes in the kidney. Parametric studies in which the cooling rate during freezing and the concentration of glycerol in the tissue were varied confirmed the results of earlier freeze-substitution studies. The results suggest a mechanism for ice propagation in the kidney similar to that already proposed for the liver, in which ice originates in, and is subsequently propagated through, the peritubular vasculature. The ice front dehydrates the cells and tubular structures encountered in its path, thus preventing intraluminal freezing. At higher rates of cooling and increased concentrations of glycerol there is less dehydration of cortical structure and intraluminal freezing occurs.

Animals↗

[The nature of the sense of effort and its neural substrate].

INTRODUCTION: What are the nature and the neural substrate of voluntary force perception? STATE OF ART: Experimental findings demonstrate that efferent signals related to motor command play a dominant role in perceiving voluntary muscular force. This suggests that voluntary force perception is provided through a sense of effort and not through a sense of intramuscular tension. Nevertheless, experimental data show that the contribution of sensory input to effort awareness must not be dismissed. Sensory signals are not involved in generating a signal of effort but rather in calibrating and modulating its magnitude. Neuroimaging and neuropsychological studies revealed that many cortical structures are activated during tasks of voluntary muscular force perception. PERSPECTIVES AND CONCLUSION: In such tasks, the basal ganglia might support the coherence of cortical activity.

Basal Ganglia↗

Mindboggle: automated brain labeling with multiple atlases.

BACKGROUND: To make inferences about brain structures or activity across multiple individuals, one first needs to determine the structural correspondences across their image data. We have recently developed Mindboggle as a fully automated, feature-matching approach to assign anatomical labels to cortical structures and activity in human brain MRI data. Label assignment is based on structural correspondences between labeled atlases and unlabeled image data, where an atlas consists of a set of labels manually assigned to a single brain image. In the present work, we study the influence of using variable numbers of individual atlases to nonlinearly label human brain image data. METHODS: Each brain image voxel of each of 20 human subjects is assigned a label by each of the remaining 19 atlases using Mindboggle. The most common label is selected and is given a confidence rating based on the number of atlases that assigned that label. The automatically assigned labels for each subject brain are compared with the manual labels for that subject (its atlas). Unlike recent approaches that transform subject data to a labeled, probabilistic atlas space (constructed from a database of atlases), Mindboggle labels a subject by each atlas in a database independently. RESULTS: When Mindboggle labels a human subject's brain image with at least four atlases, the resulting label agreement with coregistered manual labels is significantly higher than when only a single atlas is used. Different numbers of atlases provide significantly higher label agreements for individual brain regions. CONCLUSION: Increasing the number of reference brains used to automatically label a human subject brain improves labeling accuracy with respect to manually assigned labels. Mindboggle software can provide confidence measures for labels based on probabilistic assignment of labels and could be applied to large databases of brain images.

Journal Article↗

Brain perfusion spect in juvenile neuro-Behçet's disease.

UNLABELLED: Regional cerebral blood flow was evaluated by (99m)Tc-hexamethylpropyleneamine oxime SPECT in 7 patients (age range, 7--18 y; mean age, 9.1 y) affected with Behçet's disease and signs or symptoms of central nervous system involvement at different times of their clinical history. METHODS: Three patients suffered from seizures, 3 patients were affected with severe persistent headache that was refractory to common analgesic and nonsteroidal antiinflammatory drugs, and 1 patient had recurrent episodes of acute intracranial hypertension. Electroencephalography was performed on all patients, MRI on 5 patients, and CT on 1 patient. Brain SPECT was performed using a high-resolution, brain-dedicated camera. After conventional visual analysis by 2 expert readers, 2 transaxial sections were drawn parallel to the bicommissural line: the first across the thalami and the second across the temporal lobe at the level of the mesiotemporal structures. Cortical regions of interest were drawn automatically on the cortical ribbon on the 2 sections, whereas other regions of interest were drawn by hand around the basal ganglia, the thalami, and the mesiotemporal structures. Asymmetry analysis was then applied, and hypoperfusion was considered when the asymmetry value was >10%. RESULTS: Hypoperfusion was observed in all patients by visual and asymmetry analyses; this finding was localized mainly in the basal ganglia, the thalami, and the temporal cortex, including its mesial portion. Temporal hypoperfusion was found primarily in patients with seizures, and hypoperfusion of deep gray nuclei was found mainly in the other patients. Electroencephalography disclosed brain functional impairment in 5 of 6 patients, where- as MRI showed multiple bilateral white matter lesions in 1 patient suffering from persistent headache. CONCLUSION: As in adults, perfusion SPECT seems to be very sensitive in disclosing brain abnormalities in children and adolescents with Behçet's disease and signs or symptoms of central nervous system involvement, even with negative findings on brain MRI.

Behcet Syndrome↗

[Effects of experimental factors on the tinctorial properties of structural elements of the mammalian cerebral cortex (intravital microscopic study)].

Effects of a vital dye--methylene blue--on brain neuronal structures were investigated in the alive preparations of mammal brains by means of contact optical microscope in falling polarization light. Tinctorial properties of neuronal cortical structures were studied under the influence of a number of experimental factors (illumination conditions, dye concentration, temperature regime and oxygen supply to the preparation). A certain dependence in the character of dyeing and bleaching neuronal cells on the factors mentioned above was stated. Conditions for appearance of the alternating photodynamical effect and measures to prevent it were clarified. Practical recommendations were given for application methylene blue in prevital morphological investigations of mammal central nervous system.

Animals↗

Deep brain stimulation for Parkinson's disease dissociates mood and motor circuits: a functional MRI case study.

Behavioral disturbances have been reported with subthalamic (STN) deep brain stimulation (DBS) treatment in Parkinson's disease (PD). We report correlative functional imaging (fMRI) of mood and motor responses induced by successive right and left DBS. A 36-year-old woman with medically refractory PD and a history of clinically remitted depression underwent uncomplicated implantation of bilateral STN DBS. High-frequency stimulation of the left electrode improved motor symptoms. Unexpectedly, right DBS alone elicited several reproducible episodes of acute depressive dysphoria. Structural and functional magnetic resonance imaging (fMRI) imaging was carried out with sequential individual electrode stimulation. The electrode on the left was within the inferior STN, whereas the right electrode was marginally superior and lateral to the intended STN target within the Fields of Forel/zona incerta. fMRI image analysis (Analysis of Functional NeuroImages, AFNI) contrasting OFF versus ON stimulation identified significant lateralized blood oxygen level-dependent (BOLD) signal changes with DBS (P < 0.001). Left DBS primarily showed changes in motor regions: increases in premotor and motor cortex, ventrolateral thalamus, putamen, and cerebellum as well as decreases in sensorimotor/supplementary motor cortex. Right DBS showed similar but less extensive change in motor regions. More prominent were the unique increases in superior prefrontal cortex, anterior cingulate (Brodmann's area [BA] 24), anterior thalamus, caudate, and brainstem, and marked widespread decreases in medial prefrontal cortex (BA 9/10). The mood disturbance resolved spontaneously in 4 weeks despite identical stimulation parameters. Transient depressive mood induced by subcortical DBS stimulation was correlated with changes in mesolimbic cortical structures. This case provides new evidence supporting cortical segregation of motor and nonmotor cortico-basal ganglionic systems that may converge in close proximity at the level of the STN and the adjacent white matter tracts (Fields of Forel/zona incerta).

Adult↗

Consistent EEG focalities detected in subjects with primary generalized epilepsies monitored for two decades.

PURPOSE: To describe the evolution of interictal findings in serial EEGs from patients with primary generalized epilepsy. METHODS: A cohort of 89 subjects with various primary generalized epilepsies were reviewed. Thirty-one did not meet a priori criteria. Of the 58 patients analyzed, 12 had only absence seizures, 28 had absence seizures followed by one or more generalized tonic-clonic seizures, 9 had generalized tonic-clonic seizures followed by absence and/or myoclonic seizures. and 9 had juvenile myoclonic epilepsy. Patients were followed for a mean of 16 years. An average of 39 EEGs were obtained on each patient. RESULTS: Thirty-two patients (56%) had focal features present in up to 65% of the EEGs in each of the patients. Accepted focalities were only those that were consistent in lateralization, location and, often, morphology across the span of the study. Focal findings were most often temporal or frontal. CONCLUSIONS: Patients with typical primary generalized epilepsies show a high incidence of focal EEG features that cannot be explained on the basis of structural lesions, coincidental factors, or to artifacts of the selection criteria. Although the data do not allow a definitive explanation, possible mechanisms include associated focal cortical pathology such as microdysgenesis, and development over time of localized, self-sustaining hyperexcitability in low-threshold cortical structures subjected to repeated generalized spike-wave activity. Either hypothesis implies the participation and interaction of genetic, ontogenic, and environmental factors.

Adolescent↗

Spatial organization and fine structure of the cortical filament layer in normal locomoting Amoeba proteus.

The fine structural organization of a cortical filament layer in normal locomoting Amoeba proteus was demonstrated using improved fixation and embedding techniques. Best results were obtained after application of PIPES-buffered glutaraldehyde in connection with substances known to prevent the depolymerization of F-actin, followed by careful dehydration and freeze-substitution. The filament layer is continuous along the entire surface; it exhibits a varying thickness depending on the cell polarity, measuring several nm in advancing regions and 0.5-1 micron in retracting ones. Two different types of filaments are responsible for the construction of the layer: randomly distributed thin (actin) filaments forming an unordered meshwork beneath the plasma membrane, and thick (myosin) filaments mostly restricted to the uroid region in close association with F-actin. The cortical filament layer generates the motive force for amoeboid movement by contraction at posterior cell regions and induces a pressure flow that continues between the uroid with a high hydrostatic pressure and advancing pseudopodia with low one. The local destabilization of the cell surface as a precondition for the formation of pseudopodia is enabled by the detachment of the cortical filament layer from the plasma membrane. This results in morphological changes by the active separation of peripheral hyaloplasmic and central granuloplasmic regions.

Amoeba↗

Locus coeruleus neurons have axons that branch to the forebrain and cerebellum.

Utilizing a recently introduced double retrograde axonal tracing technique, the existence of collateralized axonal projections of locus coeruleus neurons to both forebrain and cerebellar cortical structures has been examined in adult mice. Following paired injection strategies using horseradish peroxidase and radiolabeled wheat germ agglutinin within portions of the forebrain and cerebellar cortices, substantial numbers of double labeled 'branched' neurons have been found that reside within all regions of the nucleus.

Afferent Pathways↗