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In Vivo Calcium Imaging of Lateral-line Hair Cells in Larval Zebrafish
摘要： Sensory hair cells are mechanoreceptors found in the inner ear that are required for hearing and balance. Hair cells are activated in response to sensory stimuli that mechanically deflect apical protrusions called hair bundles. Deflection opens mechanotransduction (MET) channels in hair bundles, leading to an influx of cations, including calcium. This cation influx depolarizes the cell and opens voltage-gated calcium channels located basally at the hair-cell presynapse. In mammals, hair cells are encased in bone, and it is challenging to functionally assess these activities in vivo. In contrast, larval zebrafish are transparent and possess an externally located lateral-line organ that contains hair cells. These hair cells are functionally and structurally similar to mammalian hair cells and can be functionally assessed in vivo. This article outlines a technique that utilizes a genetically encoded calcium indicator (GECI), GCaMP6s, to measure stimulus-evoked calcium signals in zebrafish lateral-line hair cells. GCaMP6s can be used, along with confocal imaging, to measure in vivo calcium signals at the apex and base of lateral-line hair cells. These signals provide a real-time, quantifiable readout of both mechanosensation- and presynapse-dependent calcium activities within these hair cells. These calcium signals also provide important functional information regarding how hair cells detect and transmit sensory stimuli. Overall, this technique generates useful data about relative changes in calcium activity in vivo. It is less well-suited for quantification of the absolute magnitude of calcium changes. This in vivo technique is sensitive to motion artifacts. A reasonable amount of practice and skill are required for proper positioning, immobilization, and stimulation of larvae. Ultimately, when properly executed, the protocol outlined in this article provides a powerful way to collect valuable information about the activity of hair-cells in their natural, fully integrated states within a live animal.
关键词： sensory neuroscience,Issue 141,calcium imaging,GCaMP,Neuroscience,genetically encoded indicators,hair cells,in vivo imaging,Zebrafish,lateral line,confocal imaging
更新于2025-09-04 15:30:14
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Fully implantable optoelectronic systems for battery-free, multimodal operation in neuroscience research
摘要： Recently developed ultrasmall, fully implantable devices for optogenetic neuromodulation eliminate the physical tethers associated with conventional set-ups and avoid the bulky head-stages and batteries found in alternative wireless technologies. The resulting systems allow behavioural studies without motion constraints and enable experiments in a range of environments and contexts, such as social interactions. However, these devices are purely passive in their electronic design, thereby precluding any form of active control or programmability; independent operation of multiple devices, or of multiple active components in a single device, is, in particular, impossible. Here we report optoelectronic systems that, through developments in integrated circuit and antenna design, provide low-power operation, and position- and angle-independent wireless power harvesting, with full user-programmability over individual devices and collections of them. Furthermore, these integrated platforms have sizes and weights that are not significantly larger than those of previous, passive systems. Our results qualitatively expand options in output stabilization, intensity control and multimodal operation, with broad potential applications in neuroscience research and, in particular, the precise dissection of neural circuit function during unconstrained behavioural studies.
关键词： wireless,optogenetics,neuroscience,optoelectronic systems,implantable
更新于2025-09-04 15:30:14
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All the light that we can see: a new era in miniaturized microscopy
摘要： One major challenge in neuroscience is to uncover how defined neural circuits in the brain encode, store, modify, and retrieve information. Meeting this challenge comprehensively requires tools capable of recording and manipulating the activity of intact neural networks in naturally behaving animals. Head-mounted miniature microscopes are emerging as a key tool to address this challenge. Here we discuss recent work leading to the miniaturization of neural imaging tools, the current state of the art in this field, and the importance and necessity of open-source options. We finish with a discussion on what the future may hold for miniature microscopy.
关键词： open-source,imaging tools,neuroscience,neural circuits,head-mounted miniature microscopes
更新于2025-09-04 15:30:14