RESUMEN
We propose a novel, scalable, and accurate method for detecting neuronal ensembles from a population of spiking neurons. Our approach offers a simple yet powerful tool to study ensemble activity. It relies on clustering synchronous population activity (population vectors), allows the participation of neurons in different ensembles, has few parameters to tune and is computationally efficient. To validate the performance and generality of our method, we generated synthetic data, where we found that our method accurately detects neuronal ensembles for a wide range of simulation parameters. We found that our method outperforms current alternative methodologies. We used spike trains of retinal ganglion cells obtained from multi-electrode array recordings under a simple ON-OFF light stimulus to test our method. We found a consistent stimuli-evoked ensemble activity intermingled with spontaneously active ensembles and irregular activity. Our results suggest that the early visual system activity could be organized in distinguishable functional ensembles. We provide a Graphic User Interface, which facilitates the use of our method by the scientific community.
Asunto(s)
Red Nerviosa/fisiología , Células Ganglionares de la Retina/fisiología , Animales , Simulación por Computador , Electrodos , Modelos Neurológicos , Análisis de Componente Principal , Células Ganglionares de la Retina/citologíaRESUMEN
Accumulation of amyloid-beta (Aß) peptides is regarded as the hallmark of neurodegenerative alterations in the brain of Alzheimer's disease (AD) patients. In the eye, accumulation of Aß peptides has also been suggested to be a trigger of retinal neurodegenerative mechanisms. Some pathological aspects associated with Aß levels in the brain are synaptic dysfunction, neurochemical remodeling and glial activation, but these changes have not been established in the retina of animals with Aß accumulation. We have employed the Octodon degus in which Aß peptides accumulated in the brain and retina as a function of age. This current study investigated microglial morphology, expression of PSD95, synaptophysin, Iba-1 and choline acetyltransferase (ChAT) in the retina of juvenile, young and adult degus using immunolabeling methods. Neurotransmitters glutamate and gamma-aminobutyric acid (GABA) were detected using immunogold labeling and glutamate receptor subunits were quantified using Western blotting. There was an age-related increase in presynaptic and a decrease in post-synaptic retinal proteins in the retinal plexiform layers. Immunolabeling showed changes in microglial morphology characteristic of intermediate stages of activation around the optic nerve head (ONH) and decreasing activation toward the peripheral retina. Neurotransmitter expression pattern changed at juvenile ages but was similar in adults. Collectively, the results suggest that microglial activation, synaptic remodeling and neurotransmitter changes may be consequent to, or parallel to Aß peptide and phosphorylated tau accumulation in the retina.
RESUMEN
Although the properties of the neurons of the visual system that process central and peripheral regions of the visual field have been widely researched in the visual cortex and the LGN, they have scarcely been documented for the retina. The retina is the first step in integrating optical signals, and despite considerable efforts to functionally characterize the different types of retinal ganglion cells (RGCs), a clear account of the particular functionality of cells with central vs. peripheral fields is still wanting. Here, we use electrophysiological recordings, gathered from retinas of the diurnal rodent Octodon degus, to show that RGCs with peripheral receptive fields (RF) are larger, faster, and have shorter transient responses. This translates into higher sensitivity at high temporal frequencies and a full frequency bandwidth when compared to RGCs with more central RF. We also observed that imbalances between ON and OFF cell populations are preserved with eccentricity. Finally, the high diversity of functional types of RGCs highlights the complexity of the computational strategies implemented in the early stages of visual processing, which could inspire the development of bio-inspired artificial systems.
RESUMEN
New studies show that the retina also undergoes pathological changes during the development of Alzheimer's disease (AD). While transgenic mouse models used in these previous studies have offered insight into this phenomenon, they do not model human sporadic AD, which is the most common form. Recently, the Octodon degus has been established as a sporadic model of AD. Degus display age-related cognitive impairment associated with Aß aggregates and phosphorylated tau in the brain. Our aim for this study was to examine the expression of AD-related proteins in young, adult and old degus retina using enzyme-linked or fluorescence immunohistochemistry and to quantify the expression using slot blot and western blot assays. Aß4G8 and Aß6E10 detected Aß peptides in some of the young animals but the expression was higher in the adults. Aß peptides were observed in the inner and outer segment of the photoreceptors, the nerve fiber layer (NFL) and ganglion cell layer (GCL). Expression was higher in the central retinal region than in the retinal periphery. Using an anti-oligomer antibody we detected Aß oligomer expression in the young, adult and old retina. Immunohistochemical labeling showed small discrete labeling of oligomers in the GCL that did not resemble plaques. Congo red staining did not result in green birefringence in any of the animals analyzed except for one old (84 months) animal. We also investigated expression of tau and phosphorylated tau. Expression was seen at all ages studied and in adults it was more consistently observed in the NFL-GCL. Hyperphosphorylated tau detected with AT8 antibody was significantly higher in the adult retina and it was localized to the GCL. We confirm for the first time that Aß peptides and phosphorylated tau are expressed in the retina of degus. This is consistent with the proposal that AD biomarkers are present in the eye.
Asunto(s)
Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/genética , Regulación de la Expresión Génica/fisiología , Octodon/metabolismo , Retina/metabolismo , Proteínas tau/genética , Péptidos beta-Amiloides/metabolismo , Animales , Modelos Animales de Enfermedad , Fosforilación , Proteínas tau/metabolismoRESUMEN
Several studies have shown that connexin channels play an important role in retinal neural coding in nocturnal rodents. However, the contribution of these channels to signal processing in the retina of diurnal rodents remains unclear. To gain insight into this problem, we studied connexin expression and the contribution of connexin channels to the retinal light response in the diurnal rodent Octodon degus (degu) compared to rat, using in vivo ERG recording under scotopic and photopic light adaptation. Analysis of the degu genome showed that the common retinal connexins present a high degree of homology to orthologs expressed in other mammals, and expression of Cx36 and Cx43 was confirmed in degu retina. Cx36 localized mainly to the outer and inner plexiform layers (IPLs), while Cx43 was expressed mostly in cells of the retinal pigment epithelium. Under scotopic conditions, the b-wave response amplitude was strongly reduced by 18-ß-glycyrrhetinic acid (ß-GA) (-45.1% in degu, compared to -52.2% in rat), suggesting that connexins are modulating this response. Remarkably, under photopic adaptation, ß-GA increased the ERG b-wave amplitude in degu (+107.2%) while reducing it in rat (-62.3%). Moreover, ß-GA diminished the spontaneous action potential firing rate in ganglion cells (GCs) and increased the response latency of ON and OFF GCs. Our results support the notion that connexins exert a fine-tuning control of the retinal light response and have an important role in retinal neural coding.
RESUMEN
Las matrices de microelectrodos (MEA) son dispositivos que permiten la detección de potenciales de acción o espigas en poblaciones de células excitables, ofreciendo varias aplicaciones en el campo de las neurociencias y la biología. Este trabajo muestra un protocolo para el registro de espigas en una población de células ganglionares retinales empleando una matriz de microelectrodos. La retina de una rata albina fue extraída y preparada para ser estimulada in vitro con luz led blanca, con el fin de registrar sus espigas evocadas ante estos estímulos. Cada microelectrodo puede registrar espigas de más de una célula ganglionar, razón por la cual se determinó a qué célula pertenece cada espiga aplicando un procedimiento conocido como "clasificación de espigas". El trabajo permitió obtener el registro de un periodo de estimulación y otro de no estimulación, con el fin de representar los potenciales de acción evocados con luz y los espontáneos. Los registros fueron almacenados para visualizar las espigas de las células ganglionares y poder aplicar la herramienta de clasificación de espigas. De este modo, se almacenan los instantes de tiempo en los cuales cada célula ganglionar registrada generó potenciales de acción. Este trabajo conllevó al establecimiento de un protocolo de experimentación básico enfocado al uso de matrices MEA en el laboratorio de adquisición de potenciales extracelulares de la Universidad Antonio Nariño Sede Bogotá, no sólo para caracterizar los potenciales de acción de células ganglionares retinales, sino también para otro tipo de células que puedan ser estudiadas empleando matrices de microelectrodos.
The microelectrode arrays (MEA) are devices that allow the detection of action potentials or spikes in populations of excitable cells, offering a wide spectrum of applications in topics of Neurosciences and Biology. This work describes a protocol for recording of spikes in a population of retinal ganglion cells employing a microelectrode array. The retina of an albino rat was dissected and prepared to be stimulated in vitro with white led light and to record their evoked spikes. Each microelectrode can record spikes from more than a ganglion cell, for which it was necessary to determine which cell fires each spike applying a procedure known as spike sorting. The work allowed to obtain the recording of a stimulation period and another of non-stimulation, representing evoked and spontaneous action potentials. The recordings were saved, in order to visualize the action potentials of the ganglion cells detected and to apply a computational method for the spike sorting. In this way, it was saved the time stamps in which each action potential was fired by its respective cell. This work established a basic experimentation protocol focused to the use of MEA devices in the laboratory for acquisition of extracellular potentials at the Antonio Nariño University - Bogota Headquarters, not only for characterization of action potentials fired by retinal ganglion cells populations, but also for other kind of cells that can be studied employing MEA devices.