Ultracompact mirror device for forming 20-nm achromatic soft-X-ray focus toward multimodal and multicolor nanoanalyses.

Nanoscale soft-X-ray microscopy is a powerful analysis tool in biological, chemical, and physical sciences. To enhance its probe sensitivity and leverage multimodal soft-X-ray microscopy, precise achromatic focusing devices, which are challenging to fabricate, are essential. Here, we develop an ultracompact Kirkpatrick-Baez (ucKB) mirror, which is ideal for the high-performance nanofocusing of broadband-energy X-rays. We apply our advanced fabrication techniques and short-focal-length strategy to realize diffraction-limited focusing over the entire soft-X-ray range. We achieve a focus size of 20.4 nm at 2 keV, which represents a significant improvement in achromatic soft-X-ray focusing. The ucKB mirror extends soft-X-ray fluorescence microscopy by producing a bicolor nanoprobe with a 1- or 2-keV photon energy. We propose a subcellular chemical mapping method that allows a comprehensive analysis of specimen morphology and the distribution of light elements and metal elements. ucKB mirrors will improve soft-X-ray nanoanalyses by facilitating photon-hungry, multimodal, and polychromatic methods, even with table-top X-ray sources.

Global remapping in granule cells and mossy cells of the mouse dentate gyrus.

Hippocampal place cells exhibit spatially modulated firing, or place fields, which can remap to encode changes in the environment or other variables. Unique among hippocampal subregions, the dentate gyrus (DG) has two excitatory populations of place cells, granule cells and mossy cells, which are among the least and most active spatially modulated cells in the hippocampus, respectively. Previous studies of remapping in the DG have drawn different conclusions about whether granule cells exhibit global remapping and contribute to the encoding of context specificity. By recording granule cells and mossy cells as mice foraged in different environments, we found that by most measures, both granule cells and mossy cells remapped robustly but through different mechanisms that are consistent with firing properties of each cell type. Our results resolve the ambiguity surrounding remapping in the DG and suggest that most spatially modulated granule cells contribute to orthogonal representations of distinct spatial contexts.

Change in network dynamics over time by administering Notch response inhibitor DAPT to hippocampal culture.

Although previous researches have investigated the relationship between learning and memory function in the hippocampus and continuously produced newborn neurons, the detailed role of newly generated neurons remains unclear. Here, we investigated the correlation between immature neurons and the electrical activity of the hippocampus at the network level in vitro. We showed that administrating the Notch response inhibitor DAPT to the hippocampal network enhances the neuronal differentiation of newborn cells and decreases the ratio of immature neurons in hippocampal culture. Unlike the hippocampal network without DAPT, the network with DAPT decreased the burst duration and the coefficient of variation of interburst intervals over culturing time and showed a higher synchronization level of the network over time. Moreover, the number of neurons playing a receiver or sender neuron was lower in the network with DAPT than without DAPT. Our results indicate that immature neurons may contribute to assigning neurons specific nodes as the receiver of the sender and to the diversity of the network activity while altering connections among neurons in the network.Clinical Relevance- Our research demonstrated the effect of DAPT on the ratio of immature neurons. Furthermore, our study showed the role of immature neurons in the hippocampus at the network level.

Modulation of dynamics in a pre-existing hippocampal network by neural stem cells on a microelectrode array.

Objective.Neural stem cells (NSCs) are continuously produced throughout life in the hippocampus, which is a vital structure for learning and memory. NSCs in the brain incorporate into the functional hippocampal circuits and contribute to processing information. However, little is known about the mechanisms of NSCs' activity in a pre-existing neuronal network. Here, we investigate the role of NSCs in the neuronal activity of a pre-existing hippocampalin vitronetwork grown on microelectrode arrays.Approach.We assessed the change in internal dynamics of the network by additional NSCs based on spontaneous activity. We also evaluated the networks' ability to discriminate between different input patterns by measuring evoked activity in response to external inputs.Main results.Analysis of spontaneous activity revealed that additional NSCs prolonged network bursts with longer intervals, generated a lower number of initiating patterns, and decreased synchronization among neurons. Moreover, the network with NSCs showed higher synchronicity in close connections among neurons responding to external inputs and a larger difference in spike counts and cross-correlations during evoked response between two different inputs. Taken together, our results suggested that NSCs alter the internal dynamics of the pre-existing hippocampal network and produce more specific responses to external inputs, thus enhancing the ability of the network to differentiate two different inputs.Significance.We demonstrated that NSCs improve the ability to distinguish external inputs by modulating the internal dynamics of a pre-existing network in a hippocampal culture. Our results provide novel insights into the relationship between NSCs and learning and memory.

Functional Scaffolding for Brain Implants: Engineered Neuronal Network by Microfabrication and iPSC Technology.

Neuroengineering methods can be effectively used in the design of new approaches to treat central nervous system and brain injury caused by neurotrauma, ischemia, or neurodegenerative disorders. During the last decade, significant results were achieved in the field of implant (scaffold) development using various biocompatible and biodegradable materials carrying neuronal cells for implantation into the injury site of the brain to repair its function. Neurons derived from animal or human induced pluripotent stem (iPS) cells are expected to be an ideal cell source, and induction methods for specific cell types have been actively studied to improve efficacy and specificity. A critical goal of neuro-regeneration is structural and functional restoration of the injury site. The target treatment area has heterogeneous and complex network topology with various types of cells that need to be restored with similar neuronal network structure to recover correct functionality. However, current scaffold-based technology for brain implants operates with homogeneous neuronal cell distribution, which limits recovery in the damaged area of the brain and prevents a return to fully functional biological tissue. In this study, we present a neuroengineering concept for designing a neural circuit with a pre-defined unidirectional network architecture that provides a balance of excitation/inhibition in the scaffold to form tissue similar to that in the injured area using various types of iPS cells. Such tissue will mimic the surrounding niche in the injured site and will morphologically and topologically integrate into the brain, recovering lost function.

Change in Evoked Response of Mature Neuronal Network to Spatial Pattern Stimulation by Immature Neurons.

Adult neurogenesis in the hippocampus is known to enhance pattern separation. However, the effect of adult neurogenesis on spatial pattern separation at the cellular assembly level is unclear. In order to elucidate how newborn and immature neurons change learning of spatial pattern of mature neuronal network, we evaluated evoked response to two types of spatial patterns of the cultured hippocampal network with or without added neural stem cells by using electrical stimulation on microelectrode array. Results show that the existence of newborn and immature neurons changed evoked response of mature neuronal network to both trained and untrained patterns, suggesting that the presence of immature neurons may contribute to production of the change that mature neuronal network enhances LTP and excitation to stimuli.