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Organotypic slice cultures for analysis of proliferation, cell death, and migration in the embryonic neocortex.

Dynamic cellular interactions during neocortical neurogenesis are critical for proper cortical development, providing both trophic and tropic support. Although cell proliferation and programmed cell death have been characterized in dissociated primary cell cultures, many in vivo processes during cortical neurogenesis depend on cell-cell interactions and therefore on the three-dimensional environment of the proliferating neuroblasts and their progeny. Here we describe a murine organotypic neocortical slice preparation that retains major morphological and functional in vivo characteristics of the developing neocortex and is viable (exhibits very low levels of cell death) for up to three days. Moreover, this slice preparation is amenable to direct experimental manipulation of potential diffusible regulators of neurogenesis. Using a variety of biochemical and physiological methods including time-lapse and quantitative confocal microscopy, we demonstrate that this system can be used effectively to investigate cellular mechanisms important for brain growth and maturation, including neurogenesis, apoptosis, and neuronal migration.

Animals↗

Signaling pathways that regulate specification of neurons in developing cerebral cortex.

The expression of the limbic system-associated membrane protein (LAMP), a marker of specific functional regions of the cerebral cortex, has been used to determine the environmental signals that regulate cortical regionalization. Transplant and cell culture studies have shown previously that the fate of precursor cells, based on LAMP expression, is amenable to regulation by exposure to novel environmental stimuli. This has been demonstrated in vitro to be dependent upon exposure to transforming factor-alpha and collagen type IV. Results following exposure to the inductive signals for a specific duration indicate a cell cycle dependence on the decision to become a limbic or nonlimbic cortical neuron. It appears, therefore, that areal and laminar fates are both influenced by mechanisms that specify commitment early in cortical development.

Animals↗

Graphical simulation of early development of the cerebral cortex.

Much experimental data exists concerning the development of the cerebral cortex. There is a need for a common vehicle to integrate this data and to allow the testing of hypotheses concerning development. Computer simulation and visualization are powerful mechanisms for hypothesis testing. Our long-term goal is to create a robust, extensible, portable tool for simulation and visualization of cortical development to serve both research and educational purposes. This paper describes a simulation program, SimCortex, which models the early stages of development of the cerebral cortex of the mouse. Version 1.0 of SimCortex models the proliferation of progenitor cells in the pseudostratified ventricular epithelium (PVE), the generation of young neurons and their migration into the cortical plate, which is the forerunner of cell layers II through VI of the adult cortex. We present an overview of the design and implementation of SimCortex, sample output of the system and a comparison of our results with experimental data. We conclude with a brief overview of proposed future enhancements of the system.

Animals↗

Voxel-by-voxel comparison of automatically segmented cerebral gray matter--A rater-independent comparison of structural MRI in patients with epilepsy.

Quantitative evaluation of MRI in patients with epilepsy can give more information than qualitative assessment. Previously developed volume-of-interest-based methods identified subtle widespread structural changes in the neocortex beyond the visualized lesions in patients with malformations of cortical development (MCD) and hippocampal sclerosis (HS) and also in MRI-negative patients with juvenile myoclonic epilepsy (JME). This study evaluates a voxel-based automated analysis of structural MRI in epilepsy. After fully automated segmentation of cerebral gray matter from structural T1-weighted, high-resolution MRI scans, we applied the automated and objective technique of statistical parametric mapping (SPM) to the analysis of gray matter of 35 control subjects, 10 patients with partial seizures and MCD, 10 patients with left temporal lobe epilepsy (TLE) and HS, 10 patients with left TLE and normal MR quantitation of the hippocampus, and 20 patients with JME. At a corrected threshold of P < 0.05, significant abnormalities were found in 3/35 controls; in all 10 patients with MCD, 6 of whom had additional lesions beyond the margins of the visualized abnormalities; in 2/10 TLE patients with HS; in 2/10 MRI-negative TLE; and in 4/20 JME patients. Group comparisons between control subjects and HS patients identified the affected left temporal lobe with an increase in gray matter in the posterior temporal lobe, but did not identify hippocampal atrophy. The group of MRI-negative TLE patients showed no abnormalities compared with control subjects. Group comparison between control subjects and JME patients identified a mesial frontal increase in gray matter. The SPM-based voxel-by-voxel comparison of gray matter distribution identified MCD and abnormalities beyond the visualized lesion in individual MCD patients. The method did not reliably identify HS in individual patients or identify abnormalities in individual MRI-negative patients with TLE or JME in a proportion larger than the chance findings in the control group. Using group comparisons, structural abnormalities in the neocortical gray matter of patients with TLE and HS were lateralized to the affected temporal lobe. In patients with JME as a group, an increase in gray matter was localized to the mesial frontal area, corroborating earlier quantitative MRI findings.

Adolescent↗

[Cortical deafness due to bilateral temporal subcortical hemorrhages associated with moyamoya disease: report of a case].

A 51-year-old male with moyamoya disease had experienced a left putaminal hemorrhage 14 years previously, in 1983. Left encephalo-duro-arterio-synangiosis (EDAS) was performed in that year. The left putaminal hemorrhage reoccurred in 1990 and in 1994. Left superficial temporal artery-middle cerebral artery anastomosis (STA-MCA anastomosis) was performed. He was symptom free after the surgery. However, he suddenly suffered from deafness in 1997, when computerized tomography (CT) revealed the right temporal subcortical hemorrhage. Deafness continued for three days. It gradually improved and completely disappeared in two months. The damage to the bilateral auditory radiation was suggested as the cause of deafness in this patient. Cortical deafness is a rare symptom that is most often associated with cerebral infarction. This may be the first report of moyamoya disease which developed cortical deafness.

Cerebral Angiography↗

A temporal continuity to the vertical organization of the human neocortex.

Radial translaminar arrays of pyramidal cells-minicolumns-are a pervasive structural motif of placental mammalian neocortex, which are anticipated in the earliest stages of cortical development by the formation of ontogenetic cell columns comprising radial glial cells and associated radially migrating neurons. In the present study we examine the temporal continuity in these structures throughout development and aging. Computerized image analysis of micrograph Nissl-stained postmortem tissue produced estimates of the median free path through neuropil in the radial direction (parallel to pyramidal cell arrays) and in the tangential direction (parallel to the cortical surface). These data were modeled as a biphasic power law with respect to in utero development and postnatal age, multiplied by a decay factor. No significant change in the ratio of radial to tangential neuropil space was demonstrated in either the prenatal or postnatal sample population. Neuropil development follows a prenatal phase of cubic volumetric growth with a postnatal phase of linear volumetric growth. The data suggest the continuity of columnar structures from early in gestation through postnatal maturation.

Adolescent↗

Excitability changes and glucose metabolism in experimentally induced focal cortical dysplasias.

Malformations of cortical development are increasingly recognized in association with severe epileptic syndromes, neuropsychological disorders and mental retardation. Several clinical and experimental studies suggest that functional consequences of cortical dysplasias are not restricted to the area of the dysplastic lesion but also involve remote brain regions. In the present study cortical malformations were induced in newborn rats at day of birth by intracerebral injection of the glutamatergic agonist ibotenate. The resulting cytoarchitectonic lesion associates neuronal depopulation of deep cortical layers, ectopic neurons in superficial layers and sulcus formation, mimicking human polymicrogyria and migration disorders. Electrophysiological recordings of evoked field potentials in slice preparations of adult animals reveal hyperexcitability in widespread cortical regions surrounding the dysplasia. Low-intensity stimulation induced epileptiform activity consisting of long-lasting, multiphasic and N-methyl-D-aspartate-dependent field responses. They appeared with high variability as all-or-none events. These widespread changes in excitability were not observed in sham-operated animals with small superficial ectopias but intact deep cortical layers, indicating that focal loss of these layers induces extended alterations in cortical connectivity and imbalance of excitation and inhibition. Restricted zones of increased excitability were also found in the forelimb and hindlimb representation cortex in sham-operated and control animals, demonstrating that this activity has to be considered as an intrinsic property of specific cortical areas. Deoxyglucose autoradiography showed that the widespread hyperexcitability in ibotenate-injected animals was not accompanied by alterations in glucose metabolism, although in the area of structural abnormality a typical metabolic pattern was found, revealing an increased glucose uptake in layer I. Hypometabolism as described for many types of human dysplastic lesions was not observed. This difference between the experimental and clinical data may be due to the absence of behavioral seizures in this model. However, it can be hypothesized that in patients with developmental malformations, additional pathogenic factors contribute to the manifestation of seizure disorders.

Animals↗

Two cases of reversible posterior leukoencephalopathy syndrome, one with and the other without pre-eclampsia.

Two cases of reversible posterior leukoencephalopathy syndrome (RPLS) are reported. One was a 26-year-old woman, who had pre-eclampsia and developed cortical blindness and subsequent eclampsia at 28 weeks' gestation. The other was a 27-year-old woman, who had no pre-eclampsia and developed loss of consciousness and subsequent systemic convulsion at 36 weeks' gestation. On brain magnetic resonance imaging, they both had high signal intensity on T2-weighted and fluid-attenuated inversion recovery images, and normal signal intensity on diffusion-weighted image of the posterior lobe, which almost disappeared with the amelioration of clinical symptoms thereafter. RPLS is considered to be the result of vasogenic brain edema caused by hypertension. Two hypotheses are conceived to explain the emergence of RPLS without hypertension. The first suggests that an immunotolerant condition such as pregnancy can easily cause vasogenic edema without the elevation of blood pressure. The second suggests that hypertension exists but cannot be detected because it is extremely acute and transient.

Adult↗

Neurodevelopmental disorders as a cause of seizures: neuropathologic, genetic, and mechanistic considerations.

This review will consider patterns of developmental neuropathologic abnormalities-malformations of cortical development (MCD)--encountered in infants (often with infantile spasms), children, and adults with intractable epilepsy. Treatment of epilepsy associated with some MCD, such as focal cortical dysplasia and tubers of tuberous sclerosis, may include cortical resection performed to remove the "dysplastic" region of cortex. In extreme situations (eg, hemimegalencephaly), hemispherectomy may be carried out on selected patients. Neuropathologic (including immunohistochemical) findings within these lesions will be considered. Other conditions that cause intractable epilepsy and often mental retardation, yet are not necessarily amenable to surgical treatment (eg, lissencephaly, periventricular nodular heterotopia, double cortex syndrome) will be discussed. Over the past 10 years there has been an explosion of information on the genetics of MCD. The genes responsible for many MCD (eg, TSC1, TSC2, LIS-1, DCX, FLN1) have been cloned and permit important mechanistic studies to be carried out with the purpose of understanding how mutations within these genes result in abnormal cortical cytoarchitecture and anomalous neuroglial differentiation. Finally, novel techniques allowing for analysis of patterns of gene expression within single cells, including neurons, is likely to provide answers to the most vexing and important question about these lesions: Why are they epileptogenic?

Brain↗

Prenatal and perinatal development of radial cell columns in the human auditory cortex.

The aim of this study was to analyse cytoarchitectonic development of radial cell columns in the human auditory cortex and to correlate these 'ontogenetic' cell columns with the 'adult' pattern of radial cell arrangement. For this cytoarchitectonic analysis, brains were obtained from human fetuses and infants ranging between 9 weeks of gestation and third postnatal month. Plastic and celloidin sections containing prospective auditory areas TC, TB and TA of Economo & Koskinas were stained by Nissl method. In youngest fetuses (8-13 weeks) radially oriented cell columns (1-3 cell wide 'ontogenetic' columns) were found in the whole thickness of the developing cortical plate. During the next developmental stage (13-26 weeks of gestation), radial cell columns were present in the superficial part of the cortical plate, while the deep part of the cortical plate showed a variable cell arrangement due to the formation of the subplate layer (13-15 weeks). The appearance of the pale bands, and development of the prospective granular layer (18-26 weeks). After 28 weeks, parallel to the intensive areal cytoarchitectonic differentiation and ingrowth of callosal afferents, there was a gradual regional rearrangement in the ontogenetic pattern of vertical cell columnation. In the area supratemporalis granulosa (TC), radial columnation was observed as increased granularity and the appearance of short cords composed of drop-like cells. In the area supratemporalis simplex (TB) there was a progressive differentiation of elongated cell columns, intervening fibrillar stripes with appearance of pyramidal cell cords in layers III ('organ pipe' formations).(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Reversible cortical blindness and convulsions with cyclosporin A toxicity in a patient undergoing allogeneic peripheral stem cell transplantation.

A 40-year-old woman underwent allogeneic peripheral blood stem cell transplantation for relapsed AML-M6. She developed graft-versus-host disease on day +15 post-transplant, for which she was treated with cyclosporin A and methyl prednisolone. On day +19 she developed cortical blindness, headache and convulsions which were associated with white matter changes on MRI scanning of the head and elevated cyclosporin A levels. A diagnosis of cyclosporin A encephalopathy was made and cyclosporin A was discontinued. Her vision recovered completely after 48 h and the other symptoms resolved. This is the first case of cyclosporin A encephalopathy to be reported in an allogeneic PBSC recipient.

Adult↗

Anatomical abnormalities in dopaminoceptive regions of the cerebral cortex of dopamine D1 receptor mutant mice.

Alteration of dopamine neurotransmission during development can induce specific changes in neuronal structure and function. Here, we report specific morphological and neurochemical changes of projection neurons and interneurons of the medial frontal cortex of the dopamine D(1) receptor null mouse. Using immunostaining of cytoskeletal proteins and a crossbred D(1) receptor null:YFP transgenic reporter line, we demonstrate that the apical dendrites of pyramidal cells are abnormally organized in the prefrontal and anterior cingulate cortices of mice lacking the D(1) receptor. Neuronal processes exhibit a decrease in bundling and an increase in irregular, tortuous patterning as they weave a course towards the pial surface. In addition, there is increased parvalbumin staining of the dendrites of cortical interneurons in D(1) receptor null mice. Both pyramidal and interneuron alterations are evident by the early postnatal period and persist into adulthood. The alterations show regional specificity, in that dendritic profiles of projection neurons and interneurons in somatosensory and visual cortices develop normally. The abnormalities are reminiscent of those induced by prenatal exposure to cocaine in rabbits, an insult which has been shown to produce an attenuation of D(1) receptor-mediated responses through G(salpha). These results suggest that loss of D(1) receptor-mediated signaling during development produces permanent alterations in the cellular organization of specific cortical areas involved in attention, cognition, and emotion. Pharmacological and behavioral studies in the D(1) null mouse should be interpreted in the context of possible altered circuitry, given the presence of these developmental defects in the organization of dopaminoceptive regions of the cerebral cortex.

Animals↗

Postnatal development of cannabinoid receptor type 1 expression in rodent somatosensory cortex.

Endocannabinoids are powerful modulators of synaptic transmission that act on presynaptic cannabinoid receptors. Cannabinoid receptor type 1 (CB1) is the dominant receptor in the CNS, and is present in many brain regions, including sensory cortex. To investigate the potential role of CB1 receptors in cortical development, we examined the developmental expression of CB1 in rodent primary somatosensory (barrel) cortex, using immunohistochemistry with a CB1-specific antibody. We found that before postnatal day (P) 6, CB1 receptor staining was present exclusively in the cortical white matter, and that CB1 staining appeared in the gray matter between P6 and P20 in a specific laminar pattern. CB1 staining was confined to axons, and was most prominent in cortical layers 2/3, 5a, and 6. CB1 null (-/-) mice showed altered anatomical barrel maps in layer 4, with enlarged inter-barrel septa, but normal barrel size. These results indicate that CB1 receptors are present in early postnatal development and influence development of sensory maps.

Age Factors↗

The mechanisms of generation and propagation of synchronized bursting in developing networks of cortical neurons.

The characteristics and mechanisms of synchronized firing in developing networks of cultured cortical neurons were studied using multisite recording through planar electrode arrays (PEAs). With maturation of the network (from 3 to 40 d after plating), the frequency and propagation velocity of bursts increased markedly (approximately from 0.01 to 0.5 Hz and from 5 to 100 mm/sec, respectively), and the sensitivity to extracellular magnesium concentration (0-10 mM) decreased. The source of spontaneous bursts, estimated from the relative delay of onset of activity between electrodes, varied randomly with each burst. Physical separation of synchronously bursting networks into several parts using an ultraviolet laser, divided synchronous bursting into different frequencies and phases in each part. Focal stimulation through the PEA was effective at multiple sites in eliciting bursts, which propagated over the network from the site of stimulation. Stimulated bursts exhibited both an absolute refractory period and a relative refractory period, in which partially propagating bursts could be elicited. Periodic electrical stimulation (at 1 to 30 sec intervals) produced slower propagation velocities and smaller numbers of spikes per burst at shorter stimulation intervals. These results suggest that the generation and propagation of spontaneous synchronous bursts in cultured cortical neurons is governed by the level of spontaneous presynaptic firing, by the degree of connectivity of the network, and by a distributed balance between excitation and recovery processes.

Animals↗

Cajal-Retzius cells, inhibitory interneuronal populations and neuropeptide Y expression in focal cortical dysplasia and microdysgenesis.

Focal cortical dysplasia (FCD) and microdysgenesis (MD) are likely to represent abnormalities of radial neuronal migration during cortical development. We investigated the distribution of reelin-positive Cajal-Retzius cells, known to be important in the later stages of radial neuronal migration and cortical organization, in 12 surgical cases of both MD and FCD. Quantitation revealed significantly higher numbers of these cells in MD cases compared to controls. As the majority of cortical interneurones arise via tangential rather than radial migration, we studied the distribution and morphology of inhibitory interneuronal subsets immunolabelled for calbindin, parvalbumin and calretinin within these malformations. Frequent findings were a reduction of inhibitory interneurones in the region of FCD and abnormally localised hypertrophic or multipolar calbindin-positive interneurones in both FCD and MD. Neuropeptide Y immunostaining showed a striking increase in the density of the superficial plexus of fibres in both MD and FCD cases in addition to labelling of dysplastic neurones, which may represent an adaptive anti-convulsant mechanism to dampen down seizure propagation.

Adolescent↗

Effect of injection of testosterone derivatives to pregnant rats on the brain of their one-day offspring.

Intramuscular injection of Sustanon-250, a drug with testosterone derivatives of various action rate and duration, to rats on day 19 of gestation affected brain development in their offspring. This effect manifested in greater brain weight and neocortex thickness, smaller density of neurons in the developing cortical layer V, and larger size of these neurons and their nuclei and cytoplasm in comparison with control neonatal rats. These data attest to accelerated cerebral development in the offspring of experimental rats in comparison offspring of control rats.

Animals↗

Profile of neurologic disorders associated with HIV/AIDS from Bangalore, south India (1989-96).

One hundred patients (95 males, 5 females, mean age at presentation 31.6 +/- 9.4 yr) with various neurological disorders associated with HIV infection during 1989-1996 were evaluated at NIMHANS, Bangalore. Eighty patients belonged to group I associated with opportunistic neuroinfections and 20 to group II--non infectious neurological disorders. Cryptococcal meningitis either alone (n = 31) or associated with tuberculous meningitis (n = 6) was the most common (46.3%) followed by neurotuberculosis either alone (n = 24) or with cerebral toxoplasmosis (n = 4) accounting for 35 per cent. Other opportunistic neuroinfections included cerebral toxoplasmosis, herpes zoster, fulminant pyogenic meningitis and neurosyphilis. Clinical characteristics, diagnostic clues, their laboratory and radiological profiles and problems encountered in diagnosis and management of these opportunistic infections are highlighted. In group II (19 males and one female; mean age of 32.6 +/- 9.4 yr), two patients had cortical dementia, three acute brain stem involvement, two epilepsy and one had features suggestive of progressive multifocal leukoencephalopathy. Two patients of group I during follow up developed cortical dementia. Six had peripheral nervous system involvement similar to Guillain-Barre syndrome. Sixty six patients (63 of group I and 3 of group II) progressed to AIDS, 33 patients from group I and one patient from group II succumbed to the disease. With the rapid increase in the incidence of HIV/AIDS and an increase in the neurological manifestations of HIV/AIDS it is important to recognise the magnitude of the problem for health planning in India.

AIDS-Related Opportunistic Infections↗

Chronic invasive monitoring for identifying seizure foci in children.

In both children and adults, it has been well established that the precise localization of a seizure focus allows for the best possible resection and outcome. Long-term invasive monitoring with DE has been the most widely used modality to evaluate patients with intractable seizures of temporal lobe origin. The study of epilepsy in children, however, requires assessment of the cause of the seizures with a resultant decision regarding the optimal modality of study for the particular problem. Complex partial seizures of medial temporal lobe origin without clear concordance or lateralizing information are best studied using DE. With MR imaging anatomic localization, stereotactic insertion of DE can be performed accurately and relatively safely. The information obtained permits lateralization and focus localization if the lesion is within the medial structures. The ability of DE to study epilepsy outside of the medial temporal lobe, however, is suboptimal. Many centers have used surface electrodes to map temporal lobe foci successfully. SE and SGE have not been widely used in children. Subdural electrode arrays, however, are superior to DE in extratemporal epilepsy because the cortical surface contacts provide the best recordings of epileptiform activity from a wide range of extratemporal regions. The limitations of intraoperative ECoG in the awake child are well understood. It is rare that a child can undergo surgery under local anesthesia and cooperate to the extent necessary for definitive mapping of the seizure focus and eloquent areas. Thus, invasive long-term monitoring would seem ideal in the epileptic pediatric patient because it allows for a more relaxed pace in which to record events, and because of the indwelling nature of these electrodes, it is possible for the investigator to map eloquent areas of the brain before resection. This type of evaluation completely and accurately maps interictal and ictal activity, and through cortical stimulation or mapping of somatosensory areas by evoked potentials, eloquent areas of the brain, such as motor and speech regions, can be identified. This method is limited to older children because the young child often has inadequate cortical development to localize these areas definitively. One should note that in young children (under the age of 4 years), the absence of a response to standard cortical stimulation does not indicate nonfunctional cortex. Long-term intractable seizures and significant antiepileptic drug requirements may contribute to developmental and psychosocial deficiencies in the epileptic patient.(ABSTRACT TRUNCATED AT 400 WORDS)

Child↗