ABSTRACT
Introducción. El trastorno por déficit de atención (TDA) ha sido estudiado desde muchos puntos de vista, sin embargo, todavía se desconocen los mecanismos neurobiológicos subyacentes al mismo. Los potenciales evocados y entre ellos el componente P300 pueden servir para investigar los procesos de las funciones cognitivas y atencionales deficitarios en los niños con TDA. Metodología. En este estudio analizamos la eficacia del programa del neuroeducativo HERVAT (acrónimo de Hidratación, Equilibrio, Respiración. Visión, Audición, Tacto) en el potencial evocado P300 en un grupo de niños, entre 7 y 11 años con TDA. Resultados. Los resultados indican que al final del estudio los niños con TDA que han hecho el programa HERVAT han mejorado la latencia del P300 y han reorganizado la actividad cerebral hacia áreas frontales mientras que el grupo control mantiene la misma latencia del P300 y las mismas áreas corticales posteriores durante la tarea de discriminación de estímulos multisensoriales. Conclusiones. Como conclusión podríamos decir que el programa neuroeducativo HERVAT manifiesta su eficacia en el acortamiento de la latencia del potencial evocado P300, responsable del procesamiento cerebral de la información, así como en la reorganización de la actividad cerebral desde áreas posteriores cerebrales hacia áreas corticales frontales, responsables de los procesos atencionales de las funciones ejecutivas
Introduction. Attention deficit disorder (ADD) has been investigated from various perspectives. However, the neurobiological mechanisms underlying this condition remain unknown. Evoked potentials, including P300, can be used to investigate the processes underlying deficient attentional and cognitive functions in children with ADD. Methods. In this study, we analyze the effect of a neuroeducational program, HERVAT (Hidratacion [hydration], Equilibrio [balance], Respiracion [breathing], Vision [vision], Audicion [hearing], Tacto [touch]), on evoked potential P300 in a group of children aged 7-11 years with ADD. Results. At the end of the study, the latency of P300 improved and brain activity was reorganized toward frontal areas in children with ADD who undertook the HERVAT program. In the control group, on the other hand, the latency of P300 and the posterior cortical areas remained unchanged during tests to discriminate between multisensory stimuli. Conclusions. In conclusion, the neuroeducational program HERVAT effectively shortened the latency of evoked potential P300, which is responsible for information processing in the brain, and reorganized brain activity from posterior areas toward frontal cortical areas, which are responsible for the attentional processes involved in executive function
Subject(s)
Humans , Male , Female , Child , Attention Deficit Disorder with Hyperactivity/physiopathology , Event-Related Potentials, P300/physiology , Cognition/physiology , Neurobiology/instrumentation , Education, Primary and Secondary , Attention Deficit Disorder with Hyperactivity/diagnosis , Electroencephalography/methodsABSTRACT
With the ability to correct for the aberrations introduced by biological specimens, adaptive optics-a method originally developed for astronomical telescopes-has been applied to optical microscopy to recover diffraction-limited imaging performance deep within living tissue. In particular, this technology has been used to improve image quality and provide a more accurate characterization of both structure and function of neurons in a variety of living organisms. Among its many highlights, adaptive optical microscopy has made it possible to image large volumes with diffraction-limited resolution in zebrafish larval brains, to resolve dendritic spines over 600µm deep in the mouse brain, and to more accurately characterize the orientation tuning properties of thalamic boutons in the primary visual cortex of awake mice.
Subject(s)
Microscopy , Neurobiology/instrumentation , Neurobiology/methods , Neurons/physiology , Optics and Photonics , Animals , Brain/cytology , Dendrites/ultrastructure , Humans , Larva , Neurons/cytology , ZebrafishSubject(s)
Nematoda/physiology , Nervous System Physiological Phenomena , Research , Animals , Caenorhabditis elegans/physiology , Drug Discovery , Nematode Infections/drug therapy , Nematode Infections/parasitology , Nematode Infections/prevention & control , Neurobiology/instrumentation , Neurobiology/trends , Review Literature as TopicABSTRACT
Ultrasound neuro-modulation has gained increasing attention as a non-invasive method. In this paper, we present an ultrasound neuro-modulation chip, capable of initiating reversal behaviour and activating neurons of C. elegans under the stimulation of a single-shot, short-pulsed ultrasound. About 85.29% ± 6.17% of worms respond to the ultrasound stimulation exhibiting reversal behaviour. Furthermore, the worms can adapt to the ultrasound stimulation with a lower acoustic pulse duration of stimulation. In vivo calcium imaging shows that the activity of ASH, a polymodal sensory neuron in C. elegans, can be directly evoked by the ultrasound stimulation. On the other hand, AFD, a thermal sensitive neuron, cannot be activated by the ultrasound stimulation using the same parameter and the temperature elevation during the stimulation process is relatively small. Consistent with the calcium imaging results, the tax-4 mutants, which are insensitive to temperature increase, do not show a significant difference in avoidance probability compared to the wild type. Therefore, the mechanical effects induced by ultrasound are the main reason for neural and behavioural modulation of C. elegans. With the advantages of confined acoustic energy on the surface, compatible with standard calcium imaging, this neuro-modulation chip could be a powerful tool for revealing the molecular mechanisms of ultrasound neuro-modulation.
Subject(s)
Acoustic Stimulation/instrumentation , Caenorhabditis elegans/radiation effects , Lab-On-A-Chip Devices , Neurobiology/instrumentation , Sensory Receptor Cells/radiation effects , Animals , Caenorhabditis elegans/metabolism , Caenorhabditis elegans/physiology , Molecular Imaging/methods , Neurobiology/methods , Sensory Receptor Cells/physiology , Ultrasonic WavesABSTRACT
Este es el primero de dos trabajos que abordan la relación entre la mentalización (MZ) y la conducta violenta. La MZ o función reflexiva es una forma de actividad mental imaginativa involucrada en la percepción e interpretación del comportamiento humano en términos de estados mentales intencionales. Se considera un mediador en la relación entre las experiencias tempranas en el sistema de apego y posteriores dificultades en la regulación afectiva, la atención y el autocontrol. Las experiencias traumáticas en la infancia son un aspecto crucial en la psicogénesis de la violencia y si se llevan a cabo por una figura de apego, interfieren en el proceso de desarrollo de la MZ. Por ello, se ha vinculado la violencia a un fallo significativo de la función reflexiva. Asimismo, se cuenta con una amplia evidencia biológica consistente con este supuesto
This is the first of two papers addressing the relationship between mentalization (MZ) and violent behavior. Mentalizing can be defined as a form of imaginative mental activity involved in the perception and interpretation of human behaviour in terms of intentional mental states. It is considered a mediator in the relationship between early attachment system experiences and subsequent difficulties in affect regulation, attention and self-control. Traumatic childhood experiences are crucial in the psychogenesis of violence and when perpetrated by an attachment figure, it is suggested that trauma interferes with the mentalization development process. Therefore, violence has been linked to a significant failure in mentalization
Aquest és el primer de dos treballs que aborden la relació entre la mentalització (MZ) i la conducta violenta. La MZ o funció reflexiva és una forma dactivitat mental imaginativa involucrada en la percepció i interpretació del comportament humà en termes destats mentals intencionals. Es considera un mediador en la relació entre les experiències primerenques en el sistema daferrament i posteriors dificultats en la regulació afectiva, latenció i lautocontrol. Les experiències traumàtiques en la infantesa són un aspecte crucial en la psicogènesi de la violència i si es porten a terme amb una figura daferrament, interfereixen en el procés de desenvolupament de la MZ. Per això, sha vinculat la violència a una fallida significativa de la funció reflexiva. Al mateix temps, es compta amb una ampla evidència biològica consistent amb aquest supòsit
Subject(s)
Humans , Male , Female , Violence/prevention & control , Violence/psychology , Theory of Mind/physiology , Behavior , Psychological Trauma/etiology , Psychological Trauma/psychology , Psychological Trauma/therapy , Aggression/psychology , Problem Behavior/psychology , Identity Crisis , Therapeutics/instrumentation , Therapeutics/methods , Antisocial Personality Disorder/psychology , Antisocial Personality Disorder/therapy , Borderline Personality Disorder/psychology , Borderline Personality Disorder/therapy , Psychopathology/instrumentation , Psychopathology/methods , Neurobiology/instrumentation , Neurobiology/methods , Mental HealthABSTRACT
Los trastornos del espectro autista son más prevalentes en los varones que en las mujeres, y la proporción puede variar desde 1,4 a 1 hasta 15,7 a 1, dependiendo de las muestras analizadas. La menor diferencia se ha relacionado con quienes manifiestan además discapacidad intelectual asociada, y se acepta que en esos casos las mujeres se afectan mucho más gravemente. Es probable que exista un subregistro de mujeres con trastorno del espectro autista, en especial en las que tienen alto rendimiento cognitivo, posiblemente relacionado con las técnicas de evaluación utilizadas e incluso con la falta de adecuados niveles de alerta en las niñas. En general, las mujeres con autismo tienen mejor desarrollo lingüístico temprano, mejores habilidades sociales y su juego puede incluso desarrollarse en la forma pretendida. Sus intereses pueden ser similares a los de su grupo de pares, aunque en general varían en intensidad y calidad. Se acepta que la diferencia en las habilidades sociales se hace más evidente en la adolescencia. La teoría del cerebro masculino extremo, el factor protector femenino, variantes en la plasticidad cerebral (menor umbral en los varones con mayor susceptibilidad) y factores genéticos y epigenéticos, entre otros, se evocan como posibles hipótesis que justifican esta menor prevalencia y las variantes clínicas en ellas. Este trabajo se propone analizar los aspectos clínicos y evolutivos, la variabilidad de expresión en las mujeres en relación con los varones, y algunas de las posibles bases neurobiológicas y genéticas que justifican la mayor prevalencia y las diferencias de expresión (AU)
Autism spectrum disorders are more prevalent in males than in females, and the proportion can range from 1.4 to 1, depending on the samples that are analysed. The smaller difference has been related to those who also manifest an associated intellectual disability, and it is accepted that in those cases females are far more seriously affected. There is likely to be a subregister of females with autism spectrum disorder, especially in those who have high cognitive performance, that is possibly related with the assessment techniques that are used and even with the lack of suitable levels of arousal in girls. In general, females with autism have better early language development, better social skills and their playing can even develop in the expected way. Their interests can be similar to those of their peer group, although they usually vary in intensity and quality. It is accepted as a fact that the difference in the social skills becomes more apparent in adolescence. The extreme male brain theory, the female-specific protective factor, variants in brain plasticity (lower threshold in males with greater susceptibility) and genetic and epigenetic factors, among others, are put forward as possible hypotheses to justify this lower prevalence and the clinical variants in females. This work aims to analyse the clinical and developmental aspects, the variability of expression in females with respect to males, and some of the possible neurobiological and genetic bases that account for the higher prevalence and the differences in expression (AU)
Subject(s)
Humans , Female , Autistic Disorder/diagnosis , Autistic Disorder/etiology , Autistic Disorder/genetics , Autism Spectrum Disorder/epidemiology , Autism Spectrum Disorder/genetics , Autism Spectrum Disorder/therapy , Sex Characteristics , Neuronal Plasticity/genetics , Neuronal Plasticity/physiology , Women , Social Communication Disorder/complications , Social Communication Disorder/diagnosis , Neurobiology/instrumentation , Neurobiology/methods , Early Diagnosis , Quality of LifeABSTRACT
Introducción. La Childrens Communication Checklist (CCC) de Bishop es una prueba útil para la valoración de la pragmática verbal en los escolares. El objetivo del trabajo es comprobar la fiabilidad y la validez de esta escala en castellano. Sujetos y métodos. Se analiza la CCC contestada por los padres de 360 niños/as de 4-12 años, con inteligencia normal, 160 controles y 200 de cuatro grupos clínicos: trastorno por déficit de atención/hiperactividad (n = 68), trastorno de aprendizaje no verbal procedimental (n = 77), trastorno de la comunicación social (n = 25) y trastornos del espectro autista de nivel 1 (n = 30). Se realizan análisis: factorial para agrupar los ítems del cuestionario, de fiabilidad de las nuevas escalas y discriminante para comprobar si clasifica bien a los afectos de dificultades en el uso del lenguaje. Resultados. Se obtienen siete factores (Kaiser-Meyer-Olkin: 0,852) con moderada similitud a las de la CCC original: relaciones sociales, intereses y otros cinco que constituyen pragmática (habilidades conversacionales, coherencia-comprensión, compenetración, comunicación no verbal y pertinencia). La correlación es significativa entre todos ellos, en el grupo control, y entre los cinco que configuran pragmática, en los grupos clínicos (r de Pearson). La fiabilidad de las escalas es buena (alfa de Cronbach: 0,914). El cuestionario clasifica bien al 98,9% de los casos agrupados con y sin trastorno pragmático; y al 78% de los participantes en sus correspondientes grupos clínicos. Además, permite diferenciar las patologías según la presencia e intensidad de los síntomas. Conclusiones. Esta versión española de la CCC es altamente válida y fiable. Los estadísticos aportados pueden utilizarse como valores de referencia (AU)
Introduction. The Childrens Communication Checklist (CCC) by Bishop is a useful scale for evaluation of pragmatic verbal abilities in school children. The aim of the study is to ascertain the validity and reliability of the CCC in Spanish. Subjects and methods. Answers to the CCC items by parents of 360 children with normal intelligence were analyzed. There were five groups: 160 control children; 68 children with attention deficit hyperactivity disorder, 77 with procedural non-verbal disorder, 25 children with social communication disorder and 30 with autism spectrum disorder. Investigations included: factorial analysis in order to cluster checklist items, reliability analyses of the proposed scales and discriminant analysis to check whether the scale correctly classifies children with pragmatic verbal abilities. Results. Seven factors were obtained (Kaiser-Meyer-Olkin: 0.852) with moderate similarity with those of the original scale: social relationships, interests, and five more that can be grouped into pragmatic verbal ability (conversational abilities, coherence-comprehension, empathy nonverbal communication and appropriateness). All factors are significantly correlated with each other in the control group, and the five that compose pragmatic verbal ability correlate with each other in the clinical groups (Pearson r). The scales have good reliability (Cronbachs alpha: 0.914). The questionnaire correctly classifies 98.9% of grouped cases with and without pragmatic disorder and 78% of subjects in their appropriate clinical group. Besides, the questionnaire allows to differentiate the pathologies according to the presence and intensity of the symptoms. Conclusions. This Spanish version of the CCC is highly valid and reliable. The proposed statistics can be used as normativereference values (AU)
Subject(s)
Humans , Male , Female , Child, Preschool , Child , 35249 , Social Communication Disorder/complications , Social Communication Disorder/diagnosis , Autism Spectrum Disorder/diagnosis , Autism Spectrum Disorder/pathology , Autism Spectrum Disorder/therapy , Autistic Disorder/pathology , Autistic Disorder/psychology , Autistic Disorder/therapy , Reproducibility of Results , Neurobiology/instrumentation , Neurobiology/methods , ComorbiditySubject(s)
Brain Mapping/methods , Animals , Brain/anatomy & histology , Brain/cytology , Brain/physiology , Brain/ultrastructure , Brain Mapping/instrumentation , Brain Mapping/standards , Brain Mapping/trends , Connectome/instrumentation , Connectome/methods , Humans , Magnetic Resonance Imaging , Mice , Microscopy, Electron, Scanning , Neural Pathways/anatomy & histology , Neural Pathways/cytology , Neural Pathways/physiology , Neural Pathways/ultrastructure , Neurobiology/instrumentation , Neurobiology/methods , Neurobiology/trends , Retina/cytologySubject(s)
Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods , Animals , Behavior, Animal , Culture Techniques/instrumentation , Culture Techniques/methods , Equipment Design , Humans , Immunohistochemistry/instrumentation , Immunohistochemistry/methods , Microscopy/instrumentation , Microscopy/methods , Neurobiology/instrumentation , Neurobiology/methodsABSTRACT
Optogenetic approaches allow the manipulation of neuronal activity patterns in space and time by light, particularly in small animals such as zebrafish. However, most techniques cannot control neuronal activity independently at different locations. Here we describe equipment and provide a protocol for single-photon patterned optical stimulation of neurons using a digital micromirror device (DMD). This method can create arbitrary spatiotemporal light patterns with spatial and temporal resolutions in the micrometer and submillisecond range, respectively. Different options to integrate a DMD into a multiphoton microscope are presented and compared. We also describe an ex vivo preparation of the adult zebrafish head that greatly facilitates optogenetic and other experiments. After assembly, the initial alignment takes about one day and the zebrafish preparation takes <30 min. The method has previously been used to activate channelrhodopsin-2 and manipulate oscillatory synchrony among spatially distributed neurons in the zebrafish olfactory bulb. It can be adapted easily to a wide range of other species, optogenetic probes and scientific applications.
Subject(s)
Light , Neurobiology/instrumentation , Neurobiology/methods , Neurons/physiology , Optical Devices , Photons , Zebrafish/physiology , Animals , Neurons/cytology , Optics and Photonics/methods , Photic StimulationABSTRACT
Durante mucho tiempo, la principal fuente de información para entender cómo se organiza el lenguaje en el cerebro fueron los estudios neuropsicológicos. Sin embargo, las técnicas avanzadas de neuroimágenes y electrofisiología proporcionan nuevos datos sobre el procesamiento ling³ístico en los cerebros intactos, mostrando que los procesos del lenguaje parecen estar mucho más ampliamente distribuidos de lo pensado hasta entonces. Este artículo revisa la información reciente sobre la organización neurobiológica de las palabras y sus significados. Los datos actuales sugieren que las palabras se extienden corticalmente como redes de neuronas con diferentes topografías corticales que reflejan su significado, o más exactamente, aspectos de su referencia. Por ejemplo, diferentes subcategorías de palabras de acción dan lugar a diferentes respuestas cerebrales. Así, encontramos que leer palabras relacionadas con el olor, tales como ôcanelaö, ôajoö o ôjazmínö, causa activación en el cortex olfativo primario, y la lectura de palabras relacionadas con el sabor, tales como ôsalö o ômielö, activa regiones cerebrales del gusto. Estos y otros datos indican que el significado de una palabra no está confinado a una región específica del cerebro; por el contrario, parece probable que las representaciones semánticas se encuentren distribuidas de un modo sistemático a través de todo el cerebro, incorporando información sensorial y motora (AU)
Subject(s)
Cerebral Cortex/anatomy & histology , Language Development , Neurobiology/instrumentation , Neurobiology/trends , Electrophysiology , /trendsABSTRACT
Durante mucho tiempo, la principal fuente de información para entender cómo se organiza el lenguaje en el cerebro fueron los estudios neuropsicológicos. Sin embargo, las técnicas avanzadas de neuroimágenes y electrofisiología proporcionan nuevos datos sobre el procesamiento lingüístico en los cerebros intactos, mostrando que los procesos del lenguaje parecen estar mucho más ampliamente distribuidos de lo pensado hasta entonces. Este artículo revisa la información reciente sobre la organización neurobiológica de las palabras y sus significados. Los datos actuales sugieren que las palabras se extienden corticalmente como redes de neuronas con diferentes topografías corticales que reflejan su significado, o más exactamente, aspectos de su referencia. Por ejemplo, diferentes subcategorías de palabras de acción dan lugar a diferentes respuestas cerebrales. Así, encontramos que leer palabras relacionadas con el olor, tales como canela, ajo o jazmín, causa activación en el cortex olfativo primario, y la lectura de palabras relacionadas con el sabor, tales como sal o miel, activa regiones cerebrales del gusto. Estos y otros datos indican que el significado de una palabra no está confinado a una región específica del cerebro; por el contrario, parece probable que las representaciones semánticas se encuentren distribuidas de un modo sistemático a través de todo el cerebro, incorporando información sensorial y motora
Subject(s)
Cerebral Cortex/anatomy & histology , Language Development , Electrophysiology , Linguistics/trends , Neurobiology/instrumentation , Neurobiology/trendsABSTRACT
Los enfoques actualmente utilizados para analizar la neurobiología molecular de trastornos neuropsiquiátricos complejos, como la esquizofrenia y la depresión mayor, han sido criticados con razón por no aportar un beneficio a los pacientes. La mejora del potencial translacional de nuestros trabajos requerirá el desarrollo y perfeccionamiento de mejores modelos de la enfermedad, que tengan en cuenta una amplia variedad de factores contribuyentes, como la variación genética, las interacciones entre gen y entorno, la determinación del endofenotipo o el fenotipo intermedio, el análisis en distintas especies, las diferencias entre sexos y las etapas del desarrollo. En el transcurso de una reunión específica de expertos del European College of Neuropsychopharmacology (ECNP), celebrada en Estambul, abordamos las oportunidades y los fallos existentes en los modelos animales translacionales actuales de los trastornos psiquiátricos y acordamos una serie de guías y recomendaciones clave que pensamos que serán útiles para orientar la ulterior investigación en este campo (AU)
Current approaches to dissect the molecular neurobiology of complex neuropsychiatric disorders such as schizophrenia and major depression have been rightly criticized for failing to provide benefits to patients. Improving the translational potential of our efforts will require the development and refinement of better disease models that consider a wide variety of contributing factors, such as genetic variation, gene-by-environment interactions, endophenotype or intermediate phenotype assessment, cross species analysis, sex differences, and developmental stages. During a targeted expert meeting of the European College of Neuropsychopharmacology (ECNP) in Istanbul, we addressed the opportunities and pitfalls of current translational animal models of psychiatric disorders and agreed on a series of core guidelines and recommendations that we believe will help guiding further research in this area (AU)
Subject(s)
Animals , Male , Female , Mice , Neurobiology/methods , Neurobiology/trends , Aging/physiology , Sex Differentiation/physiology , Mental Disorders/diagnosis , Mental Disorders/veterinary , Risk Factors , Models, Animal , Neurogenesis/physiology , Biological Psychiatry/methods , Biological Psychiatry/trends , Neurobiology/ethics , Neurobiology/instrumentation , Neurobiology/organization & administration , Cultural Characteristics , Odds Ratio , Models, Genetic , Neurogenesis/geneticsABSTRACT
Advances in neural tissue engineering have resulted in the development and implementation of three-dimensional (3-D) neural cellular constructs, which may serve as neurofidelic in vitro investigational platforms. In addition, interfacing these 3-D cellular constructs with micro-fluidic and/or micro-electrical systems has created biohybridized platforms, providing unprecedented 3-D microenvironmental control and allowing noninvasive probing and manipulation of cultured neural cells. Cells in the brain interact within a complex, multicellular environment with tightly coupled 3-D cell-cell/cell-extracellular matrix (ECM) interactions; yet most in vitro models utilize planar systems lacking in vivo-like ECM. As such, neural cultures with cells distributed throughout a thick (> 500 microm), bioactive extracellular matrix may provide a more physiologically relevant setting to study neurobiological phenomena than traditional planar cultures. This review presents an overview of 2-D versus 3-D culture models and the state of the art in 3-D neural cell-culture systems. We then detail our efforts to engineer a range of 3-D neural cellular constructs by systematically varying parameters such as cell composition, cell density, matrix constituents, and mass transport. The ramifications on neural cell survival, function, and network formation based on these parameters are specifically addressed. These 3-D neural cellular constructs may serve as powerful investigational platforms for the study of basic neurobiology, network neurophysiology, injury/disease mechanisms, pharmacological screening, or test-beds for cell replacement therapies. Furthermore, while survival and growth of neural cells within 3-D constructs poses many challenges, optimizing in vitro constructs prior to in vivo implementation offers a sound bioengineering design approach.
Subject(s)
Cell Culture Techniques , Microfluidic Analytical Techniques/methods , Neurobiology/methods , Neurons/cytology , Neurons/physiology , Tissue Engineering/methods , Animals , Cell Survival , Coculture Techniques , Microfluidic Analytical Techniques/instrumentation , Neurobiology/instrumentation , Phenotype , Tissue Engineering/instrumentation , Tissue ScaffoldsABSTRACT
Time-lapse imaging is often the only way to appreciate fully the many dynamic cell movements critical to neural development. Zebrafish possess many advantages that make them the best vertebrate model organism for live imaging of dynamic development events. This review will discuss technical considerations of time-lapse imaging experiments in zebrafish, describe selected examples of imaging studies in zebrafish that revealed new features or principles of neural development, and consider the promise and challenges of future time-lapse studies of neural development in zebrafish embryos and adults.
Subject(s)
Developmental Biology , Neurobiology , Neurogenesis/physiology , Time-Lapse Imaging , Animals , Developmental Biology/instrumentation , Developmental Biology/methods , Microscopy, Confocal , Molecular Imaging , Neurobiology/instrumentation , Neurobiology/methods , Time-Lapse Imaging/trends , Zebrafish/embryologyABSTRACT
Dissociated hippocampal neuron culture has long been the model system of choice for many neuroscientists. The ability to culture dissociated hippocampal neurons from genetically modified mice provides an invaluable tool for studying many neuronal processes. In this study, we established a novel method to culture dissociated hippocampal neurons from embryonic and neonatal mice. Dissociated neurons were cultured in a microchamber between the glass coverslip and the plastic cell container without the use of glial feeder cells. Our method significantly simplifies the preparation while produces healthy and long-lived neuronal cultures that are difficult to achieve without the use of feeder cells.
Subject(s)
Hippocampus/cytology , Neurobiology/instrumentation , Neurobiology/methods , Neurons/cytology , Animals , Animals, Newborn , Cell Culture Techniques/instrumentation , Cell Culture Techniques/methods , Cells, Cultured , Female , Hippocampus/physiology , Male , Mice , Mice, Inbred C57BL , Neurons/physiology , PregnancyABSTRACT
Reconstructing neuronal circuits at the level of synapses is a central problem in neuroscience, and the focus of the nascent field of connectomics. Previously used to reconstruct the C. elegans wiring diagram, serial-section transmission electron microscopy (ssTEM) is a proven technique for the task. However, to reconstruct more complex circuits, ssTEM will require the automation of image processing. We review progress in the processing of electron microscopy images and, in particular, a semi-automated reconstruction pipeline deployed at Janelia Farm. Drosophila circuits underlying identified behaviors are being reconstructed in the pipeline with the goal of generating a complete Drosophila connectome.