We present a unique, extensive, and open synaptic physiology analysis platform and dataset. Through its application, we reveal principles that relate cell type to synaptic properties and intralaminar circuit organization in the mouse and human cortex. The dynamics of excitatory synapses align with the postsynaptic cell subclass, whereas inhibitory synapse dynamics partly align with presynaptic cell subclass but with considerable overlap. Synaptic properties are heterogeneous in most subclass-to-subclass connections. The two main axes of heterogeneity are strength and variability. Cell subclasses divide along the variability axis, whereas the strength axis accounts for substantial heterogeneity within the subclass. In the human cortex, excitatory-to-excitatory synaptic dynamics are distinct from those in the mouse cortex and vary with depth across layers 2 and 3.

Generating a comprehensive description of cortical networks requires a large-scale, systematic approach. To that end, we have begun a pipeline project using multipatch electrophysiology, supplemented with two-photon optogenetics, to characterize connectivity and synaptic signaling between classes of neurons in adult mouse primary visual cortex (V1) and human cortex. We focus on producing results detailed enough for the generation of computational models and enabling comparison with future studies. Here, we report our examination of intralaminar connectivity within each of several classes of excitatory neurons. We find that connections are sparse but present among all excitatory cell classes and layers we sampled, and that most mouse synapses exhibited short-term depression with similar dynamics. Synaptic signaling between a subset of layer 2/3 neurons, however, exhibited facilitation. These results contribute to a body of evidence describing recurrent excitatory connectivity as a conserved feature of cortical microcircuits.

Key points• In mouse models for retinal degeneration, photoreceptor death leads to membrane oscillation in the remnant AII amacrine-ON cone bipolar cell network through an unknown mechanism.• We found such oscillations require voltage-gated Na + channels and gap junctions but not hyperpolarization-activated currents (I h ).• Na + channels are expressed predominantly in AII amacrine cells and I h in ON cone bipolar cells, and appear to interact via gap junctions to shape oscillations.• Similar intrinsic oscillations arose in the wild-type (wt) AII amacrine-ON cone bipolar cell network when photoreceptor inputs to bipolar cells were pharmacologically occluded.• Computational modelling captures experimental findings when a low level of cellular heterogeneity is introduced in the coupled network.• These unique insights into the cellular mechanisms underlying spontaneous activity in the degenerating retina might aid in designing the most effective strategies to restore vision using retinal prosthesis. AbstractIn the rd1 mouse model for retinal degeneration, the loss of photoreceptors results in oscillatory activity (∼10-20 Hz) within the remnant electrically coupled network of retinal ON cone bipolar and AII amacrine cells. We tested the role of hyperpolarization-activated currents (I h ), voltage-gated Na + channels and gap junctions in mediating such oscillatory activity. Blocking I h (1 mM Cs + ) hyperpolarized the network and augmented activity, while antagonizing voltage-dependent Na + channels (1 μM TTX) abolished oscillatory activity in the AII amacrine-ON cone bipolar cell network. Voltage-gated Na + channels were only observed in AII amacrine cells, implicating these cells as major drivers of activity. Pharmacologically uncoupling the network (200 μM meclofenamic acid (MFA)) blocked oscillations in all cells indicating that Na + channels exert their influence over multiple cell types within the network. In wt retina, occluding photoreceptor inputs to bipolar cells (10 μM NBQX and 50 μM L-AP4) resulted in a mild (∼10 mV) hyperpolarization and the induction of oscillatory activity within the AII amacrine-ON cone bipolar cell network. These oscillations had similar properties to those observed in rd1 retina, suggesting that no major degeneration-induced network rewiring is required to trigger spontaneous oscillations. Finally, we constructed a simplified computational model that exhibited Na + channel-dependent network oscillations. In this model, mild heterogeneities in channel densities between individual neurons reproduced our experimental findings. These results indicate that TTX-sensitive Na + channels in AII amacrine cells trigger degeneration-induced network oscillations, which provide a persistent synaptic drive to downstream remnant neurons, thus appearing to replace photoreceptors as the principal drivers of retinal activity.

A surprisingly large number of neurons throughout the brain are endowed with the ability to co-release both a fast excitatory and inhibitory transmitter. The computational benefits of dual transmitter release, however, remain poorly understood. Here, we address the role of co-transmission of acetylcholine (ACh) and GABA from starburst amacrine cells (SACs) to direction-selective ganglion cells (DSGCs). Using a combination of pharmacology, optogenetics, and linear regression methods, we estimated the spatiotemporal profiles of GABA, ACh, and glutamate receptor-mediated synaptic activity in DSGCs evoked by motion. We found that ACh initiates responses to motion in natural scenes or under low-contrast conditions. In contrast, classical glutamatergic pathways play a secondary role, amplifying cholinergic responses via NMDA receptor activation. Furthermore, under these conditions, the network of SACs differentially transmits ACh and GABA to DSGCs in a directional manner. Thus, mixed transmission plays a central role in shaping directional responses of DSGCs.

Understanding cortical function will require a detailed and comprehensive knowledge of local circuit properties. The Allen Institute for Brain Science is beginning a large-scale project using multipatch electrophysiology, supplemented with 2-photon optogenetics, to characterize local connectivity and synaptic signaling between major classes of neurons in the adult mouse primary visual cortex and neurosurgical samples from human frontal and temporal cortex. We focus on generating results that are detailed enough for the generation of computational models and enable rigorous comparison with future studies. Here we report our examination of the intralaminar connectivity within each of several classes of excitatory neurons. We find that connections are sparse but present among all excitatory cell types and layers we sampled, with the most sparse connections in layers 5 and 6. Almost all synapses in mouse exhibited short-term depression with similar dynamics. Synaptic signaling between a subset of layer 2/3 neurons, however, exhibited facilitation. These results contribute to a body of evidence describing recurrent excitatory connectivity as a conserved feature of cortical microcircuits.

We present a unique, extensive, public synaptic physiology dataset. The dataset contains over 20,000 neuron pairs probed with multipatch using standardized protocols to capture short-term dynamics. Recordings were made in the human temporal cortex and the adult mouse visual cortex. Our main purpose is to offer data and analyses that provide a more complete picture of the cortical microcircuit to the community. We also make several important findings that relate connectivity and synaptic properties to the major cell subclasses and cortical layer via the development of novel analysis methods for quantifying connectivity, synapse properties, and synaptic dynamics. We find that excitatory synaptic dynamics depend strongly on the postsynaptic cell subclass, whereas inhibitory synaptic dynamics depend on the presynaptic cell subclass. Despite these associations, short-term synaptic plasticity is heterogeneous in most subclass to subclass connections. We also find that intralaminar connection probability exhibits a strong layer dependence. In human cortex, we find that excitatory synapses are highly reliable, recover rapidly, and are distinct from mouse excitatory synapses.

Local and global forms of inhibition controlling directionally selective ganglion cells (DSGCs) in the mammalian retina are well documented. It is established that local inhibition arising from GABAergic starburst amacrine cells (SACs) strongly contributes to direction selectivity. Here, we demonstrate that increasing ambient illumination leads to the recruitment of GABAergic wide-field amacrine cells (WACs) endowing the DS circuit with an additional feature: size selectivity. Using a combination of electrophysiology, pharmacology, and light/electron microscopy, we show that WACs predominantly contact presynaptic bipolar cells, which drive direct excitation and feedforward inhibition (through SACs) to DSGCs, thus maintaining the appropriate balance of inhibition/excitation required for generating DS. This circuit arrangement permits high-fidelity direction coding over a range of ambient light levels, over which size selectivity is adjusted. Together, these results provide novel insights into the anatomical and functional arrangement of multiple inhibitory interneurons within a single computational module in the retina.

No abstract