Highly attenuated dendritic propagation of isolated synaptic potentials in vivo.
The integration of synaptic inputs is a fundamental function of neurons. In the traditional model, excitatory inputs are summed at the soma to generate action potentials. However, how synaptic inputs are integrated by dendrites in vivo remains poorly explored. We used intravital two-photon dendritic imaging with a genetically encoded voltage indicator (accelerated sensor of action potentials 5) together with somatic whole-cell patch clamp recordings to investigate how synaptic depolarizations are transferred to the soma in pyramidal neurons of the mouse somatosensory cortex. We studied the integration of synaptic inputs under spontaneous and sensory-evoked conditions, as well as following electrical and optogenetic stimulation. In all cases, while multiple inputs evoked measurable depolarizations in the cell body, isolated synaptic potentials were strongly attenuated. Our results suggest that isolated synaptic inputs have a minimal contribution to somatic depolarization, whereas coincident inputs within short temporal windows are more effective, indicating a regime of dendritic integration that favors coincident or clustered neuronal activity in cortical networks.
Voltage imaging of neurons distributed across entire brains of larval zebrafish.
Neurons interact in networks distributed throughout the brain. While much effort has focused on whole-brain calcium imaging, advances in genetically encoded voltage indicators raise the question of whether it might be possible to image neuronal voltage across entire brains. Achieving this requires a microscope with high volumetric imaging rates and signal-to-noise ratio. Here we present a remote-scanning light-sheet microscope capable of imaging genetically encoded voltage indicator-expressing neurons distributed throughout much of the brain of larval zebrafish at a volumetric rate of 200.8 Hz. We measured voltage traces from approximately one-quarter of all brain neurons. We found that neurons firing at different times during a sequence occupied different locations: visually evoked sequences mapped across the optic tectum, whereas stimulus-independent bursts were mapped across the cerebellum and medulla. Imaging voltage of neurons distributed in many brain regions may open new frontiers for understanding fundamental neural system operations.
Sub-millivolt voltage imaging reveals gap junction-mediated bioelectric contact inhibition.
Sub-millivolt membrane potential (V) dynamics in multicellular non-excitable networks have remained largely inaccessible due to insufficiently sensitive imaging tools. Here, we introduce rEstus2s, a next-generation genetically encoded voltage indicator that overcomes this barrier by enabling high-resolution V imaging. Using rEstus2s, we uncover bioelectric contact inhibition (BCI), a biophysical principle in which gap junction coupling passively stabilizes V by suppressing electrical volatility. We show that V variance scales inversely with network size (1/n), reflecting a transition from stochastic single-cell behavior to collective electrical stability. Ca²⁺-activated oncogenic ion channels, including ANO1 and K3.1, drive pronounced electrical volatility in isolated cells, but BCI effectively attenuates this volatility in electrically coupled networks. Disruption of gap junction coupling abolishes BCI and restores high electrical volatility. These findings establish a unifying framework for how multicellular systems maintain electrical homeostasis and reveal gap junction coupling as a key determinant of bioelectric stability in health and disease.
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Voltage imaging of neurons distributed across entire brains of larval zebrafish.
Neurons interact in networks distributed throughout the brain. While much effort has focused on whole-brain calcium imaging, advances in genetically encoded voltage indicators raise the question of whether it might be possible to image neuronal voltage across entire brains. Achieving this requires a microscope with high volumetric imaging rates and signal-to-noise ratio. Here we present a remote-scanning light-sheet microscope capable of imaging genetically encoded voltage indicator-expressing neurons distributed throughout much of the brain of larval zebrafish at a volumetric rate of 200.8 Hz. We measured voltage traces from approximately one-quarter of all brain neurons. We found that neurons firing at different times during a sequence occupied different locations: visually evoked sequences mapped across the optic tectum, whereas stimulus-independent bursts were mapped across the cerebellum and medulla. Imaging voltage of neurons distributed in many brain regions may open new frontiers for understanding fundamental neural system operations.
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