Robo1 regulates the development of major axon tracts and interneuron migration in the forebrain

The Slit genes encode secreted ligands that regulate axon branching, commissural axon pathfinding and neuronal migration. The principal identified receptor for Slit is Robo ( Roundabout in Drosophila). To investigate Slit signalling in forebrain development, we generated Robo1 knockout mice by targeted deletion of exon 5 of the Robo1 gene. Homozygote knockout mice died at birth, but prenatally displayed major defects in axon pathfinding and cortical interneuron migration. Axon pathfinding defects included dysgenesis of the corpus callosum and hippocampal commissure, and abnormalities in corticothalamic and thalamocortical targeting. Slit2 and Slit1/2 double mutants display malformations in callosal development, and in corticothalamic and thalamocortical targeting, as well as optic tract defects. In these animals, corticothalamic axons form large fasciculated bundles that aberrantly cross the midline at the level of the hippocampal and anterior commissures, and more caudally at the medial preoptic area. Such phenotypes of corticothalamic targeting were not observed in Robo1 knockout mice but, instead, both corticothalamic and thalamocortical axons aberrantly arrived at their respective targets at least 1 day earlier than controls. By contrast, in Slit mutants, fewer thalamic axons actually arrive in the cortex during development. Finally, significantly more interneurons ( up to twice as many at E12.5 and E15.5) migrated into the cortex of Robo1 knockout mice, particularly in both rostral and parietal regions, but not caudal cortex. These results indicate that Robo1 mutants have distinct phenotypes, some of which are different from those described in Slit mutants, suggesting that additional ligands, receptors or receptor partners are likely to be involved in Slit/Robo signalling.

The Wnt receptor Ryk is required for Wnt5a-mediated axon guidance on the contralateral side of the corpus callosum

Ryk (receptor related to tyrosine kinase) has been shown to be a novel Wnt receptor in both Caenorhabditis elegans and Drosophila melanogaster. Recently, Ryk-Wnt interactions were shown to guide corticospinal axons down the embryonic mouse spinal cord. Here we show that, in Ryk-deficient mice, cortical axons project aberrantly across the major forebrain commissure, the corpus callosum. Many mouse mutants have been described in which loss-of-function mutations result in the inability of callosal axons to cross the midline, thereby forming Probst bundles on the ipsilateral side. In contrast, loss of Ryk does not interfere with the ability of callosal axons to cross the midline but impedes their escape from the midline into the contralateral side. Therefore, Ryk(-/-) mice display a novel callosal guidance phenotype. We also show that Wnt5a acts as a chemorepulsive ligand for Ryk, driving callosal axons toward the contralateral hemisphere after crossing the midline. In addition, whereas callosal axons do cross the midline in Ryk(-/-) embryos, they are defasciculated on the ipsilateral side, indicating that Ryk also promotes fasciculation of axons before midline crossing. In summary, this study expands the emerging role for Wnts in axon guidance and identifies Ryk as a key guidance receptor in the establishment of the corpus callosum. Our analysis of Ryk function further advances our understanding of the molecular mechanisms underlying the formation of this important commissure.

Molecular control of cell type diversity in the developing spinal cord

1, During embryonic development, a diverse array of neurons and glia are generated at specific positions along the dorsoventral and rostro-caudal axes of the spinal cord from a common pool of precursor cells. 2. This cell type diversity can be distinguished by the spatially and temporally coordinated expression of several transcription factors that are also linked to cell type specification at a very early stage of spinal cord development. 3, Recent studies have started to uncover that the generation of cell type diversity in the developing spinal cord. Moreover, distinct cell types in the spinal cord appear to be determined by the spatially and temporally coordinated expression of transcription factors. 4. The expression of these factors also appears to be controlled by gradients of factors expressed by ventral and dorsal midline cells, namely Sonic hedgehog and members of the transforming growth factor-beta family. 5, Changes in the competence of precursor cells and local cell interactions may also play important roles in cell type specification within the developing spinal cord.

AVALIAÇÃO POR MEIO DE MODELOS DE GESSO DAS ALTERAÇÕES DENTOESQUELÉTICAS, PROMOVIDAS PELA EXPANSÃO RÁPIDA DA MAXILA ASSISTIDA CIRURGICAMENTE (ERMAC)

Este estudo avaliou as alterações produzidas nos arcos dentais superiores de pacientes submetidos à Expansão Rápida da Maxila Assistida Cirurgicamente (ERMAC). A amostra utilizada foi composta de 50 modelos de gesso superiores de 18 pacientes, sendo seis do sexo masculino e 12 do sexo feminino, com média de idade de 23,3 anos. Para cada paciente foram preparados três modelos de gesso obtidos em diferentes fases: Inicial, antes do procedimento operatório (T1); três meses pós-expansão (travamento do expansor) e momento da remoção do aparelho expansor tipo Hyrax e colocação da placa removível de acrílico para contenção (T2); seis meses pós-expansão e momento de remoção da placa de acrílico (T3). O dispositivo expansor utilizado foi o disjuntor tipo Hyrax. O procedimento cirúrgico adotado foi a osteotomia lateral da maxila sem o envolvimento da lâmina pterigóide, osteotomia da espinha nasal à linha média dental (incisivos centrais superiores), separação da sutura palatina mediana por meio de cinzel e separação do septo nasal. O início da ativação ocorreu no terceiro dia pós-operatório, sendo ¼ de volta pela manhã e ¼ à noite, sendo que as ativações seguiram critérios clínicos para o controle da expansão. As medidas foram realizadas por meio da máquina de medição tridimensional (SAC), baseando-se nas alterações nos três planos (vertical, sagital e transversal) que ocorreram nos modelos de gesso. Concluiu-se que: 1. Houve um aumento estatisticamente significante nas distâncias transversais em todos os grupos de dentes (de incisivos centrais até segundos molares) de T1 para T2, demonstrando a efetividade do tratamento. De T2 para T3 não houve diferença estatisticamente significante para nenhuma variável, indicando, assim, estabilidade após seis meses do término da ERMAC; 2. Houve um aumento estatisticamente significante nas inclinações dos primeiros e segundos molares dos lados direito e esquerdo e dos segundos pré-molares apenas do lado esquerdo, sugerindo um comportamento assimétrico dos dentes avaliados; 3. Houve um aumento na largura palatina nos intervalos analisados, com diferenças estatisticamente significantes entre T1 x T2 e T1 x T3; 4. Não foram observadas diferenças estatisticamente significantes na profundidade palatina nos intervalos analisados.(AU)

Avaliação de assimetria nas arcadas dentárias de indivíduos com oclusão normal e má oclusão de Classe II

A assimetria das arcadas dentárias constitui um assunto de grande importância estudado por profissionais de Ortodontia na elaboração de um diagnóstico correto, planejamento e execução de um tratamento ortodôntico bem sucedido. O objetivo deste estudo foi avaliar o grau de assimetria das arcadas dentárias em indivíduos com oclusão normal e má oclusão de Classe II, divisão 1 e 2, bem como o dimorfismo sexual existente. Foram avaliados 180 pares de modelos de estudo de indivíduos do sexo masculino e feminino, na faixa etária de 12 a 21 anos, divididos em 3 grupos de 60 pares de modelos, de acordo com a má oclusão. Os grupos foram classificados em: Grupo 1 - indivíduos com oclusão normal, Grupo 2 - indivíduos com má oclusão de Classe II divisão 1 (Cl II 1), e Grupo 3 - indivíduos com má oclusão de Classe II divisão 2 (Cl II 2). Os modelos foram medidos utilizando-se um aparelho inédito, idealizado e fabricado exclusivamente para a análise de assimetria das arcadas dentárias. Para a análise de assimetria foram realizadas 2 medições angulares desvio de linha média (DLM), posicionamento dos caninos (PC) e 3 lineares distância dos caninos à rafe palatina (DC), distância inter-caninos (DIC), posicionamento dos primeiros molares no sentido ântero-posterior (PM). Concluiu-se que a ocorrência de assimetria nas arcadas dentárias independe da má oclusão. O Grupo 1 apresentou um menor grau de assimetria nas arcadas dentárias em relação aos grupos 2 e 3, os quais apresentaram um grau de assimetria semelhante. O grau de assimetria nas arcadas dentárias inferiores, nos 3 grupos, foi maior do que nas arcadas dentárias superiores. A direção do desvio da linha média apresentou uma correlação inversamente proporcional do lado em que o molar se apresentava mesializado, nas arcadas superior e inferior dos três grupos, com exceção da arcada superior no Grupo 2 (Classe II divisão 1). Houve dimorfismo sexual estatisticamente significante para algumas medidas, porém é importante considerar os baixos valores e a disposição, destas diferenças, entre as medidas realizadas, a qual revela ter se tratado de dados obtidos ao acaso.

Functional interconnectivity between the globus pallidus and the subthalamic nucleus in the mouse brain slice

In accordance with its central role in basal ganglia circuitry, changes in the rate of action potential firing and pattern of activity in the globus pallidus (GP)-subthalamic nucleus (STN) network are apparent in movement disorders. In this study we have developed a mouse brain slice preparation that maintains the functional connectivity between the GP and STN in order to assess its role in shaping and modulating bursting activity promoted by pharmacological manipulations. Fibre-tract tracing studies indicated that a parasagittal slice cut 20 deg to the midline best preserved connectivity between the GP and the STN. IPSCs and EPSCs elicited by electrical stimulation confirmed connectivity from GP to STN in 44/59 slices and from STN to GP in 22/33 slices, respectively. In control slices, 74/76 (97%) of STN cells fired tonically at a rate of 10.3 ± 1.3 Hz. This rate and pattern of single spiking activity was unaffected by bath application of the GABAA antagonist picrotoxin (50 μM, n = 9) or the glutamate receptor antagonist (6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX) 10 μM, n = 8). Bursting activity in STN neurones could be induced pharmacologically by application of NMDA alone (20 μM, 3/18 cells, 17%) but was more robust if NMDA was applied in conjunction with apamin (20-100 nM, 34/77 cells, 44%). Once again, neither picrotoxin (50 μM, n = 5) nor CNQX (10 μM, n = 5) had any effect on the frequency or pattern of the STN neurone activity while paired STN and GP recordings of tonic and bursting activity show no evidence of coherent activity. Thus, in a mouse brain slice preparation where functional GP-STN connectivity is preserved, no regenerative synaptically mediated activity indicative of a dynamic network is evident, either in the resting state or when neuronal bursting in both the GP and STN is generated by application of NMDA/apamin. This difference from the brain in Parkinson's disease may be attributed either to insufficient preservation of cortico-striato-pallidal or cortico-subthalamic circuitry, and/or an essential requirement for adaptive changes resulting from dopamine depletion for the expression of network activity within this tissue complex. © The Physiological Society 2005.

Visual evoked electrical and magnetic response to half-field stimulation using pattern reversal stimulation

The visual evoked magnetic response to half-field stimulation using pattern reversal was studied using a d.c. SQUID coupled to a second order gradiometer. The main component of the magnetic response consisted of a positive wave at around 100 ms (P100M). At the time this component was present the response to half-field stimulation consisted of an outgoing magnetic field contralateral and extending to the midline. When the left half field was stimulated the outgoing field was over the posterior right visual cortex and when the right half field was stimulated it was over the left anterior visual cortex. These findings would correctly identify a source located in the contralateral visual cortex. The orientation of the dipoles was not that previously assumed to explain the paradoxical lateralization of the visual evoked potential. The results are discussed in terms of both electrical and magnetic models of the calcarine fissure. © 1992.

Topographic mapping and source localization of the pattern onset visual evoked magnetic response

The topography of the visual evoked magnetic response (VEMR) to a pattern onset stimulus was investigated using 4 check sizes and 3 contrast levels. The pattern onset response consists of three early components within the first 200ms, CIm, CIIm and CIIIm. The CIIm is usually of high amplitude and is very consistent in latency within a subject. Half field (HF) stimuli produce their strongest response over the contralateral hemisphere; the RHF stimulus exhibiting a lower positivity (outgoing field) and an upper negativity (ingoing field), rotated towards the midline. LHF stimulation produced the opposite response, a lower negative and an upper positive. Larger check sizes produce a single area of ingoing and outgoing field while smaller checks produce on area of ingoing and outgoing field over each hemisphere. Latency did not appear to vary with change in contrast but amplitudes increased with increasing contrast. A more detailed topographic study incorporating source localisation procedures suggested a source for CIIm - 4cm below the scalp, close to the midline with current flowing towards the lateral surface. Similar depth and position estimates but with opposite polarity were obtained for the pattern shift P100m previously. Hence, the P100m and the CIIm may originate in similar areas of visual cortex but reveal different aspects of visual processing. © 1992 Human Sciences Press, Inc.

Visual evoked electrical and magnetic response to half-field stimulation using pattern reversal stimulation

The visual evoked magnetic response to half-field stimulation using pattern reversal was studied using a dc-SQUID coupled to a second-order gradiometer. The main component of the magnetic response consisted of a positive wave at around 100ms (P100M). At the same time this component was present the reponse to half-field stimulation consisted of an outgoing field contralateral and extending to the midline. When the left half-field was stimulates the outgoing field was over the posterior right visual cortex and when the right half field was stimulated it was over the left anterior visual cortex. These findings would correltly identify a source located in the contralateral visual cortex. The orientation of the dipoles was not that previously assumed to explain the paradoxical lateralization of the visual evoked potential. The results are discussed in terms of both electrical and magnetic models of the calcarine fissure.

Topography of visual evoked magnetic responses to pattern shift, pattern onset and flash stimuli

Different visual stimuli may activate separate channels in the visual system and produce magnetic responses from the human bran which originate from distinct regions of the visual cortex. To test this hypothesis, we have investigated the distribution of visual evoked magnetic responses to three distinct visual stimuli over the occipital region of the scalp with a DC-SQUID second-order gradiometer in an ubshielded environment. Patterned stimuli were presented full field and to the right half field, while a flash stimulus was presented full field only, in five normal subjects. Magnetic responses were recorded from 20 to 42 positions over the occipital scalp. Topographic maps were prepared of the major positive component within the first 150ms to the three stimuli, i.e., the P100m (pattern shift), C11m (pattern onset) and P2m (flash). For the pattern shift stimulus the data suggested the source of the P100m was close to the midline with the current directed towards the medial surface. The data for the pattern onset C11m suggested a source at a similar depth but with the current directed away from the midline towards the lateral surface. The flash P2m appeared to originate closer to the surface of the occipital pole than both the patterned stimuli. Hence the pattern shift (which may represent movement), and the pattern onset C11m (representing contrast and contour) appear to originate in similar areas of brain but to represent different asepcts of cortical processing. By contrast, the flash P2m (representing luminance change) appears to originate in a distinct area of visual cortex closer to the occipital pole.

Automated assessment of visual fields and their inter-relation to evoked potentials in visual disorders

The Octopus Automated Perimeter was validated in a comparative study and found to offer many advantages in the assessment of the visual field. The visual evoked potential was investigated in an extensive study using a variety of stimulus parameters to simulate hemianopia and central visual field defects. The scalp topography was recorded topographically and a technique to compute the source derivation of the scalp potential was developed. This enabled clarification of the expected scalp distribution to half field stimulation using different electrode montages. The visual evoked potential following full field stimulation was found to be asymmetrical around the midline with a bias over the left occiput particularly when the foveal polar projections of the occipital cortex were preferentially stimulated. The half field response reflected the distribution asymmetry. Masking of the central 3° resulted in a response which was approximately symmetrical around the midline but there was no evidence of the PNP-complex. A method for visual field quantification was developed based on the neural representation of visual space (Drasdo and Peaston 1982) in an attempt to relate visual field depravation with the resultant visual evoked potentials. There was no form of simple, diffuse summation between the scalp potential and the cortical generators. It was, however, possible to quantify the degree of scalp potential attenuation for M-scaled full field stimuli. The results obtained from patients exhibiting pre-chiasmal lesions suggested that the PNP-complex is not scotomatous in nature but confirmed that it is most likely to be related to specific diseases (Harding and Crews 1982). There was a strong correlation between the percentage information loss of the visual field and the diagnostic value of the visual evoked potential in patients exhibiting chiasmal lesions.

The development of calibration and adaptive filtering procedures for neuromagnetic studies of human visual function

This thesis first considers the calibration and signal processing requirements of a neuromagnetometer for the measurement of human visual function. Gradiometer calibration using straight wire grids is examined and optimal grid configurations determined, given realistic constructional tolerances. Simulations show that for gradiometer balance of 1:104 and wire spacing error of 0.25mm the achievable calibration accuracy of gain is 0.3%, of position is 0.3mm and of orientation is 0.6°. Practical results with a 19-channel 2nd-order gradiometer based system exceed this performance. The real-time application of adaptive reference noise cancellation filtering to running-average evoked response data is examined. In the steady state, the filter can be assumed to be driven by a non-stationary step input arising at epoch boundaries. Based on empirical measures of this driving step an optimal progression for the filter time constant is proposed which improves upon fixed time constant filter performance. The incorporation of the time-derivatives of the reference channels was found to improve the performance of the adaptive filtering algorithm by 15-20% for unaveraged data, falling to 5% with averaging. The thesis concludes with a neuromagnetic investigation of evoked cortical responses to chromatic and luminance grating stimuli. The global magnetic field power of evoked responses to the onset of sinusoidal gratings was shown to have distinct chromatic and luminance sensitive components. Analysis of the results, using a single equivalent current dipole model, shows that these components arise from activity within two distinct cortical locations. Co-registration of the resulting current source localisations with MRI shows a chromatically responsive area lying along the midline within the calcarine fissure, possibly extending onto the lingual and cuneal gyri. It is postulated that this area is the human homologue of the primate cortical area V4.

Evaluation of hemifield sector analysis protocol in multifocal visual evoked potential (MFVEP) objective perimetry for the diagnosis and early detection of glaucomatous field defects

Visual field assessment is a core component of glaucoma diagnosis and monitoring, and the Standard Automated Perimetry (SAP) test is considered up until this moment, the gold standard of visual field assessment. Although SAP is a subjective assessment and has many pitfalls, it is being constantly used in the diagnosis of visual field loss in glaucoma. Multifocal visual evoked potential (mfVEP) is a newly introduced method used for visual field assessment objectively. Several analysis protocols have been tested to identify early visual field losses in glaucoma patients using the mfVEP technique, some were successful in detection of field defects, which were comparable to the standard SAP visual field assessment, and others were not very informative and needed more adjustment and research work. In this study, we implemented a novel analysis approach and evaluated its validity and whether it could be used effectively for early detection of visual field defects in glaucoma. OBJECTIVES: The purpose of this study is to examine the effectiveness of a new analysis method in the Multi-Focal Visual Evoked Potential (mfVEP) when it is used for the objective assessment of the visual field in glaucoma patients, compared to the gold standard technique. METHODS: 3 groups were tested in this study; normal controls (38 eyes), glaucoma patients (36 eyes) and glaucoma suspect patients (38 eyes). All subjects had a two standard Humphrey visual field HFA test 24-2 and a single mfVEP test undertaken in one session. Analysis of the mfVEP results was done using the new analysis protocol; the Hemifield Sector Analysis HSA protocol. Analysis of the HFA was done using the standard grading system. RESULTS: Analysis of mfVEP results showed that there was a statistically significant difference between the 3 groups in the mean signal to noise ratio SNR (ANOVA p<0.001 with a 95% CI). The difference between superior and inferior hemispheres in all subjects were all statistically significant in the glaucoma patient group 11/11 sectors (t-test p<0.001), partially significant 5/11 (t-test p<0.01) and no statistical difference between most sectors in normal group (only 1/11 was significant) (t-test p<0.9). sensitivity and specificity of the HAS protocol in detecting glaucoma was 97% and 86% respectively, while for glaucoma suspect were 89% and 79%. DISCUSSION: The results showed that the new analysis protocol was able to confirm already existing field defects detected by standard HFA, was able to differentiate between the 3 study groups with a clear distinction between normal and patients with suspected glaucoma; however the distinction between normal and glaucoma patients was especially clear and significant. CONCLUSION: The new HSA protocol used in the mfVEP testing can be used to detect glaucomatous visual field defects in both glaucoma and glaucoma suspect patient. Using this protocol can provide information about focal visual field differences across the horizontal midline, which can be utilized to differentiate between glaucoma and normal subjects. Sensitivity and specificity of the mfVEP test showed very promising results and correlated with other anatomical changes in glaucoma field loss.

The role of hemifield sector analysis in multifocal visual evoked potential objective perimetry in the early detection of glaucomatous visual field defects

Objective: The purpose of this study was to examine the effectiveness of a new analysis method of mfVEP objective perimetry in the early detection of glaucomatous visual field defects compared to the gold standard technique. Methods and patients: Three groups were tested in this study; normal controls (38 eyes), glaucoma patients (36 eyes), and glaucoma suspect patients (38 eyes). All subjects underwent two standard 24-2 visual field tests: one with the Humphrey Field Analyzer and a single mfVEP test in one session. Analysis of the mfVEP results was carried out using the new analysis protocol: the hemifield sector analysis protocol. Results: Analysis of the mfVEP showed that the signal to noise ratio (SNR) difference between superior and inferior hemifields was statistically significant between the three groups (analysis of variance, P<0.001 with a 95% confidence interval, 2.82, 2.89 for normal group; 2.25, 2.29 for glaucoma suspect group; 1.67, 1.73 for glaucoma group). The difference between superior and inferior hemifield sectors and hemi-rings was statistically significant in 11/11 pair of sectors and hemi-rings in the glaucoma patients group (t-test P<0.001), statistically significant in 5/11 pairs of sectors and hemi-rings in the glaucoma suspect group (t-test P<0.01), and only 1/11 pair was statistically significant (t-test P<0.9). The sensitivity and specificity of the hemifield sector analysis protocol in detecting glaucoma was 97% and 86% respectively and 89% and 79% in glaucoma suspects. These results showed that the new analysis protocol was able to confirm existing visual field defects detected by standard perimetry, was able to differentiate between the three study groups with a clear distinction between normal patients and those with suspected glaucoma, and was able to detect early visual field changes not detected by standard perimetry. In addition, the distinction between normal and glaucoma patients was especially clear and significant using this analysis. Conclusion: The new hemifield sector analysis protocol used in mfVEP testing can be used to detect glaucomatous visual field defects in both glaucoma and glaucoma suspect patients. Using this protocol, it can provide information about focal visual field differences across the horizontal midline, which can be utilized to differentiate between glaucoma and normal subjects. The sensitivity and specificity of the mfVEP test showed very promising results and correlated with other anatomical changes in glaucomatous visual field loss. The intersector analysis protocol can detect early field changes not detected by the standard Humphrey Field Analyzer test. © 2013 Mousa et al, publisher and licensee Dove Medical Press Ltd.

Acute mobile phone operation affects neural function in humans

OBJECTIVES: Mobile phones (MP) are used extensively and yet little is known about the effects they may have on human physiology. There have been conflicting reports regarding the relation between MP use and the electroencephalogram (EEG). The present study suggests that this conflict may be due to methodological differences such as exposure durations, and tests whether exposure to an active MP affects EEG as a function of time. METHODS: Twenty-four subjects participated in a single-blind fully counterbalanced cross-over design, where both resting EEG and phase-locked neural responses to auditory stimuli were measured while a MP was either operating or turned off. RESULTS: MP exposure altered resting EEG, decreasing 1-4 Hz activity (right hemisphere sites), and increasing 8-12 Hz activity as a function of exposure duration (midline posterior sites). MP exposure also altered early phase-locked neural responses, attenuating the normal response decrement over time in the 4-8 Hz band, decreasing the response in the 1230 Hz band globally and as a function of time, and increasing midline frontal and lateral posterior responses in the 30-45 Hz band. CONCLUSIONS: Active MPs affect neural function in humans and do so as a function of exposure duration. The temporal nature of this effect may contribute to the lack of consistent results reported in the literature.