The diverse roles of inhibition in identified neural circuits

Inhibitory interneurons represent a diverse population of cell types in the central nervous system, whose general role is to suppress activity of target neurons. The timing of spikes in principal neurons has millisecond precision, and I asked what are the roles of inhibition in shaping the temporal codes that emerge from different parallel local neural circuits. First I investigated the local circuitry of melanopsin-containing ganglion cells in the mouse retina, which are intrinsically photosensitive and responsible for circadian photoentrainment. Using transsynaptic viral tracing, I identified three types of melanopsin-containing ganglion cell, and found that inhibitory (GABAergic) dopaminergic amacrine cells are presynaptic to one of these types. These results provided a direct circuitry link between the medium time scale process of light-dark adaptation, which involves dopamine, and the longer time scale of the circadian rhythm. Next I characterised a subpopulation of genetically-identified neurons in the mouse retina, in order to compare the precise timing of inhibition in different circuits at a high temporal resolution. I identified eight physiologically and morphologically distinct ganglion cell types and found that each circuit could be described by a 'motif' that represented the inhibitory-excitatory interactions that lead to cell-type-specific firing patterns. The cell would fire only when the change in excitation was faster than the change in inhibition. Therefore the role of inhibition is to detect 'irrelevance' in the visual scene, only allowing the ganglion cell to fire at specific time points relating to functions that are both parallel and complementary to the other cell types. Finally, I looked deeper within the neural circuitry of one of the genetically-identified cell types, to study the mechanism of 'fast inhibition' in detecting approaching objects. Through two-photon targeted paired recordings of postsynaptic ganglion cells and presynaptic amacrine cells, I found evidence that the AII amacrine cell - a well-characterised glycinergic inhibitory interneuron known to be involved in night vision circuits - conveys fast inhibitory information to the ganglion cell via an electrical synapse with an excitatory neuron of day vision circuitry only during non-approach motion. Therefore, it appears that the role of inhibition is to dynamically interact with direct excitatory neural pathways during 'irrelevant' stimulation, suppressing or completely blocking activity, resulting in precisely timed spikes that occur in the brief moments when excitation changes faster than inhibition

Approach sensitivity in the retina processed by a multifunctional neural circuit

The detection of approaching objects, such as looming predators, is necessary for survival. Which neurons and circuits mediate this function? We combined genetic labeling of cell types, two-photon microscopy, electrophysiology and theoretical modeling to address this question. We identify an approach-sensitive ganglion cell type in the mouse retina, resolve elements of its afferent neural circuit, and describe how these confer approach sensitivity on the ganglion cell. The circuit's essential building block is a rapid inhibitory pathway: it selectively suppresses responses to non-approaching objects. This rapid inhibitory pathway, which includes AII amacrine cells connected to bipolar cells through electrical synapses, was previously described in the context of night-time vision. In the daytime conditions of our experiments, the same pathway conveys signals in the reverse direction. The dual use of a neural pathway in different physiological conditions illustrates the efficiency with which several functions can be accommodated in a single circuit

Genetic Reactivation of Cone Photoreceptors Restores Visual Responses in Retinitis Pigmentosa

Retinitis pigmentosa refers to a diverse group of hereditary diseases that lead to incurable blindness, affecting two million people worldwide. As a common pathology, rod photoreceptors die early, whereas light-insensitive, morphologically altered cone photoreceptors persist longer. It is unknown if these cones are accessible for therapeutic intervention. Here, we show that expression of archaebacterial halorhodopsin in light-insensitive cones can substitute for the native phototransduction cascade and restore light sensitivity in mouse models of retinitis pigmentosa. Resensitized photoreceptors activate all retinal cone pathways, drive sophisticated retinal circuit functions (including directional selectivity), activate cortical circuits, and mediate visually guided behaviors. Using human ex vivo retinas, we show that halorhodopsin can reactivate light-insensitive human photoreceptors. Finally, we identified blind patients with persisting, light-insensitive cones for potential halorhodopsin-based therapy

Genetically timed, activity-sensor and rainbow transsynaptic viral tools.

We developed retrograde, transsynaptic pseudorabies viruses (PRVs) with genetically encoded activity sensors that optically report the activity of connected neurons among spatially intermingled neurons in the brain. Next we engineered PRVs to express two differentially colored fluorescent proteins in a time-shifted manner to define a time period early after infection to investigate neural activity. Finally we used multiple-colored PRVs to differentiate and dissect the complex architecture of brain regions

Behavior-Dependent Activity and Synaptic Organization of Septo-hippocampal GABAergic Neurons Selectively Targeting the Hippocampal CA3 Area

Rhythmic medial septal (MS) GABAergic input coordinates cortical theta oscillations. However, the rules of innervation of cortical cells and regions by diverse septal neurons are unknown. We report a specialized population of septal GABAergic neurons, the Teevra cells, selectively innervating the hippocampal CA3 area bypassing CA1, CA2, and the dentate gyrus. Parvalbumin-immunopositive Teevra cells show the highest rhythmicity among MS neurons and fire with short burst duration (median, 38 ms) preferentially at the trough of both CA1 theta and slow irregular oscillations, coincident with highest hippocampal excitability. Teevra cells synaptically target GABAergic axo-axonic and some CCK interneurons in restricted septo-temporal CA3 segments. The rhythmicity of their firing decreases from septal to temporal termination of individual axons. We hypothesize that Teevra neurons coordinate oscillatory activity across the septo-temporal axis, phasing the firing of specific CA3 interneurons, thereby contributing to the selection of pyramidal cell assemblies at the theta trough via disinhibition