Biology subjects

Tsien, R. W.

Publications and source records attributed to Tsien, R. W..

5 recordsLinked to original sources

bioRxiv2022

Brief synaptic inhibition persistently interrupts firing of fast-spiking interneurons

Neurons perform input-output operations that integrate synaptic inputs with intrinsic electrical properties, operations generally constrained by the brevity of synaptic events. Here we report that sustained firing of CA1 hippocampal fast-spiking parvalbumin-expressing interneurons (PV-INs) can be persistently interrupted for up to several hundred milliseconds following brief GABAAR-mediated inhibition in vitro and in vivo. A single presynaptic neuron could interrupt PV-INs firing, occasionally with a single action potential (AP), and reliably with AP bursts. Experiments and computational modeling revealed that the persistent interruption of firing maintains neurons in a depolarized, quiescent state through a cell-autonomous mechanism. Strikingly, interrupted PV-INs are highly responsive to Schaffer collateral inputs. The persistent interruption of firing provides a disinhibitory circuit mechanism favoring spike generation in CA1 pyramidal cells. Overall, our results demonstrate that neuronal silencing can far outlast brief synaptic inhibition owing to well-tuned interplay between neurotransmitter release and postsynaptic membrane dynamics, a phenomenon impacting microcircuit function.

neuroscience↗

bioRxiv2022

Synaptic homeostasis transiently leverages Hebbian mechanisms for a multiphasic response to inactivity

Neurons use various forms of negative feedback to maintain their synaptic strengths within an operationally useful range. While this homeostatic plasticity is thought to distinctly counteract the destabilizing positive feedback of Hebbian plasticity, there is considerable overlap in the molecular components mediating both forms of plasticity. The varying kinetics of these components spurs additional inquiry into the dynamics of synaptic homeostasis. We discovered that upscaling of synaptic weights in response to prolonged inactivity is nonmonotonic. Surprisingly, this seemingly oscillatory adaptation involved transient appropriation of molecular effectors associated with Hebbian plasticity, namely CaMKII, L-type Ca2+ channels, and Ca2+-permeable AMPARs, and homeostatic elements such as calcineurin. We created a dynamic model that shows how traditionally "Hebbian" and "homeostatic" mechanisms can cooperate to autoregulate postsynaptic Ca2+ levels. We propose that this combination of mechanisms allows excitatory synapses to adapt to prolonged activity changes and safeguard the capability to undergo future strengthening on demand.

neuroscience↗

bioRxiv2022

NNeurite: artificial neuronal networks for the unsupervised extraction of axonal and dendritic time-lapse signals

Fluorescence microscopy of Ca2+ transients in small neurites of the behaving mouse provides an unprecedented view of the micrometer-scale mechanisms supporting neuronal communication and computation, and therefore opens the way to understanding their role in cognition. However, the exploitation of this growing and precious experimental data is impeded by the scarcity of methods dedicated to the analysis of images of neurites activity in vivo. We present NNeurite, a set of mathematical and computational techniques specialized for the analysis of time-lapse microscopy images of neurite activity in small behaving animals. Starting from noisy and unstable microscopy images containing an unknown number of small neurites, NNeurite simultaneously aligns images, denoises signals and extracts the location and the temporal activity of the sources of Ca2+ transients. At the core of NNeurite is a novel artificial neuronal network (NN) which we have specifically designed to solve the non-negative matrix factorization (NMF) problem modeling source separation in fluorescence microscopy images. For the first time, we have embedded non-rigid image alignment in the NMF optimization procedure, hence allowing to stabilize images based on the transient and weak neurite signals. NNeurite processing is free of any human intervention as NN training is unsupervised and the unknown number of Ca2+ sources is automatically obtained by the NN-based computation of a low-dimensional representation of time-lapse images. Importantly, the spatial shapes of the sources of Ca2+ fluorescence are not constrained in NNeurite, which allowed to automatically extract the micrometer-scale details of dendritic and axonal branches, such dendritic spines and synaptic boutons, in the cortex of behaving mice. We provide NNeurite as a free and open-source library to support the efforts of the community in advancing in vivo microscopy of neurite activity.

neuroscience↗

bioRxiv2020

Unique dynamics and exocytosis of single inhibitory synaptic vesicles

Maintaining the balance between neuronal excitation and inhibition is essential for proper function of the central nervous system, with inhibitory synaptic transmission playing an important role. Although inhibitory transmission has higher kinetic demands compared to excitatory transmission, its properties are poorly understood. In particular, the dynamics and exocytosis of single inhibitory vesicles have not been investigated, due largely to both technical and practical limitations. Using a combination of quantum dots (QDs) conjugated to antibodies against the luminal domain of the vesicular GABA transporter (VGAT) to selectively label GABAergic (i.e., inhibitory) vesicles together with dual-focus imaging optics, we tracked the real-time three-dimensional position of single inhibitory vesicles up to the moment of exocytosis (i.e., fusion). Using three-dimensional trajectories, we found that inhibitory synaptic vesicles traveled a short distance prior to fusion and had a shorter time to fusion compared to synaptotagmin-1 (Syt1)-labeled vesicles, which were mostly from excitatory neurons. Moreover, our analysis revealed a close correlation between the release probability of inhibitory vesicles and both the proximity to their fusion site and the total travel length. Finally, we found that inhibitory vesicles have a higher prevalence of kiss-and-run fusion compared than Syt1-labeled vesicles. These results indicate that inhibitory synaptic vesicles have a unique set of dynamics and fusion properties to support rapid synaptic inhibition, thereby maintaining a tightly regulated balance between excitation and inhibition in the central nervous system. SignificanceDespite playing an important role in maintaining brain function, the dynamics of inhibitory synaptic vesicles are poorly understood. Here, we tracked the three-dimensional position of single inhibitory vesicles up to the moment of exocytosis in real time by loading single inhibitory vesicle with QDs-conjugated to antibodies against the luminal domain of the vesicular GABA transporter (VGAT). We found that inhibitory synaptic vesicles have a smaller total travel length before fusion, a shorter fusion time, and a higher prevalence of kiss-and-run than synaptotagmin-1-lableled vesicles. Our findings provide the first evidence that inhibitory vesicles have a unique set of dynamics and exocytosis properties to support rapid inhibitory synaptic transmission.

neuroscience↗

bioRxiv2020

MCH enables synchronized firing in the hippocamposeptal circuit to facilitate spatial memory

Neuropeptide melanin-concentrating hormone (MCH) plays important roles in the brain including control of energy homeostasis, sleep, learning and memory. However, the synaptic and circuitry mechanisms underlying MCH-mediated regulations remain largely unknown. Here, we uncover that MCH modulates the hippocampo (HP) -dorsal lateral septum (dLS) -lateral hypothalamus neural circuit to facilitate spatial learning and memory. MCH achieves this function by enhancing both excitatory and inhibitory synaptic transmission via presynaptic mechanisms. The dLS neuronal spiking activity in response to HP CA3 excitatory inputs is strongly controlled by feed-forward inhibition (FFI) mediated by both GABAA and GABAB receptors. Endogenous MCH signaling enhances Signal/Noise (S/N) ratio of dLS neurons by increase the excitatory strengths, meanwhile decrease the overall dLS excitability by enhance inhibition which reduces dLS FFI, and consequentially enables dLS neurons to fire with high fidelity with HP synaptic inputs. Our data unravel the multifaceted synaptic mechanisms of MCH in the defined HP-dLS circuitry which may contribute to learning and memory.

neuroscience↗