RESUMO
Visual virtual reality (VR) systems for head-fixed mice offer advantages over real-world studies for investigating the neural circuitry underlying behavior. However, current VR approaches do not fully cover the visual field of view of mice, do not stereoscopically illuminate the binocular zone, and leave the lab frame visible. To overcome these limitations, we developed iMRSIV (Miniature Rodent Stereo Illumination VR)-VR goggles for mice. Our system is compact, separately illuminates each eye for stereo vision, and provides each eye with an â¼180° field of view, thus excluding the lab frame while accommodating saccades. Mice using iMRSIV while navigating engaged in virtual behaviors more quickly than in a current monitor-based system and displayed freezing and fleeing reactions to overhead looming stimulation. Using iMRSIV with two-photon functional imaging, we found large populations of hippocampal place cells during virtual navigation, global remapping during environment changes, and unique responses of place cell ensembles to overhead looming stimulation.
Assuntos
Navegação Espacial , Realidade Virtual , Animais , Camundongos , Dispositivos de Proteção dos Olhos , Campos Visuais , Navegação Espacial/fisiologiaRESUMO
The use of SU-8 material in the production of neural sensors has grown recently. Despite its widespread application, a detailed systematic quantitative analysis concerning its biocompatibility in the central nervous system is lacking. In this immunohistochemical study, we quantified the neuronal preservation and the severity of astrogliosis around SU-8 devices implanted in the neocortex of rats, after a 2â¯months survival. We found that the density of neurons significantly decreased up to a distance of 20⯵m from the implant, with an averaged density decrease to 24⯱â¯28% of the control. At 20 to 40⯵m distance from the implant, the majority of the neurons was preserved (74⯱â¯39% of the control) and starting from 40⯵m distance from the implant, the neuron density was control-like. The density of synaptic contacts - examined at the electron microscopic level - decreased in the close vicinity of the implant, but it recovered to the control level as close as 24⯵m from the implant track. The intensity of the astroglial staining significantly increased compared to the control region, up to 560⯵m and 480⯵m distance from the track in the superficial and deep layers of the neocortex, respectively. Electron microscopic examination revealed that the thickness of the glial scar was around 5-10⯵m thin, and the ratio of glial processes in the neuropil was not more than 16% up to a distance of 12⯵m from the implant. Our data suggest that neuronal survival is affected only in a very small area around the implant. The glial scar surrounding the implant is thin, and the presence of glial elements is low in the neuropil, although the signs of astrogliosis could be observed up to about 500⯵m from the track. Subsequently, the biocompatibility of the SU-8 material is high. Due to its low cost fabrication and more flexible nature, SU-8 based devices may offer a promising approach to experimental and clinical applications in the future.
Assuntos
Materiais Biocompatíveis/farmacologia , Compostos de Epóxi/química , Neurônios/efeitos dos fármacos , Polímeros/química , Animais , Materiais Biocompatíveis/química , Encéfalo/patologia , Compostos de Epóxi/farmacologia , Feminino , Masculino , Microscopia Eletrônica de Varredura , Neuroglia/citologia , Neuroglia/efeitos dos fármacos , Neuroglia/metabolismo , Neuroglia/ultraestrutura , Neurônios/citologia , Neurônios/metabolismo , Neurônios/patologia , Polímeros/farmacologia , Próteses e Implantes , Ratos , Ratos WistarRESUMO
The long-term application of central nervous system implants is currently limited by the negative response of the brain tissue, affecting both the performance of the device and the survival of nearby cells. Topographical modification of implant surfaces mimicking the structure and dimensions of the extracellular matrix may provide a solution to this negative tissue response and has been shown to affect the attachment and behavior of both neurons and astrocytes. In our study, commonly used neural implant materials, silicon, and platinum were tested with or without nanoscale surface modifications. No biological coatings were used in order to only examine the effect of the nanostructuring. We seeded primary mouse astrocytes and hippocampal neurons onto four different surfaces: flat polysilicon, nanostructured polysilicon, and platinum-coated versions of these surfaces. Fluorescent wide-field, confocal, and scanning electron microscopy were used to characterize the attachment, spreading and proliferation of these cell types. In case of astrocytes, we found that both cell number and average cell spreading was significantly larger on platinum, compared to silicon surfaces, while silicon surfaces impeded glial proliferation. Nanostructuring did not have a significant effect on either parameter in astrocytes but influenced the orientation of actin filaments and glial fibrillary acidic protein fibers. Neuronal soma attachment was impaired on metal surfaces while nanostructuring seemed to influence neuronal growth cone morphology, regardless of surface material. Taken together, the type of metals tested had a profound influence on cellular responses, which was only slightly modified by nanopatterning.
Assuntos
Astrócitos/citologia , Nanoestruturas/química , Neurônios/citologia , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/ultraestrutura , Adesão Celular/efeitos dos fármacos , Contagem de Células , Movimento Celular/efeitos dos fármacos , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Células Cultivadas , Cones de Crescimento/efeitos dos fármacos , Cones de Crescimento/metabolismo , Hipocampo/citologia , Camundongos , Neurônios/efeitos dos fármacos , Neurônios/ultraestrutura , Platina/farmacologia , Silício/farmacologia , Propriedades de SuperfícieRESUMO
Neural probes designed for extracellular recording of brain electrical activity are traditionally implanted with an insertion speed between 1 µm/s and 1 mm/s into the brain tissue. Although the physical effects of insertion speed on the tissue are well studied, there is a lack of research investigating how the quality of the acquired electrophysiological signal depends on the speed of probe insertion. In this study, we used four different insertion speeds (0.002 mm/s, 0.02 mm/s, 0.1 mm/s, 1 mm/s) to implant high-density silicon probes into deep layers of the somatosensory cortex of ketamine/xylazine anesthetized rats. After implantation, various qualitative and quantitative properties of the recorded cortical activity were compared across different speeds in an acute manner. Our results demonstrate that after the slowest insertion both the signal-to-noise ratio and the number of separable single units were significantly higher compared with those measured after inserting probes at faster speeds. Furthermore, the amplitude of recorded spikes as well as the quality of single unit clusters showed similar speed-dependent differences. Post hoc quantification of the neuronal density around the probe track showed a significantly higher number of NeuN-labelled cells after the slowest insertion compared with the fastest insertion. Our findings suggest that advancing rigid probes slowly (~1 µm/s) into the brain tissue might result in less tissue damage, and thus in neuronal recordings of improved quality compared with measurements obtained after inserting probes with higher speeds.