A dark quencher genetically encodable voltage indicator (dqGEVI) exhibits high fidelity and speed.

Voltage sensing with genetically expressed optical probes is highly desirable for large-scale recordings of neuronal activity and detection of localized voltage signals in single neurons. Most genetically encodable voltage indicators (GEVI) have drawbacks including slow response, low fluorescence, or excessive bleaching. Here we present a dark quencher GEVI approach (dqGEVI) using a Förster resonance energy transfer pair between a fluorophore glycosylphosphatidylinositol-enhanced green fluorescent protein (GPI-eGFP) on the outer surface of the neuronal membrane and an azo-benzene dye quencher (D3) that rapidly moves in the membrane driven by voltage. In contrast to previous probes, the sensor has a single photon bleaching time constant of ∼40 min, has a high temporal resolution and fidelity for detecting action potential firing at 100 Hz, resolves membrane de- and hyperpolarizations of a few millivolts, and has negligible effects on passive membrane properties or synaptic events. The dqGEVI approach should be a valuable tool for optical recordings of subcellular or population membrane potential changes in nerve cells.

Synergy of direct and indirect cholinergic septo-hippocampal pathways coordinates firing in hippocampal networks.

The medial septum/diagonal band of Broca complex (MSDB) is a key structure that modulates hippocampal rhythmogenesis. Cholinergic neurons of the MSDB play a central role in generating and pacing theta-band oscillations in the hippocampal formation during exploration, novelty detection, and memory encoding. How precisely cholinergic neurons affect hippocampal network dynamics in vivo, however, has remained elusive. In this study, we show that stimulation of cholinergic MSDB neurons in urethane-anesthetized mice acts on hippocampal networks via two distinct pathways. A direct septo-hippocampal cholinergic projection causes increased firing of hippocampal inhibitory interneurons with concomitantly decreased firing of principal cells. In addition, cholinergic neurons recruit noncholinergic neurons within the MSDB. This indirect pathway is required for hippocampal theta synchronization. Activation of both pathways causes a reduction in pyramidal neuron firing and a more precise coupling to the theta oscillatory phase. These two anatomically and functionally distinct pathways are likely relevant for cholinergic control of encoding versus retrieval modes in the hippocampus.

Astrocyte Intermediaries of Septal Cholinergic Modulation in the Hippocampus.

The neurotransmitter acetylcholine, derived from the medial septum/diagonal band of Broca complex, has been accorded an important role in hippocampal learning and memory processes. However, the precise mechanisms whereby acetylcholine released from septohippocampal cholinergic neurons acts to modulate hippocampal microcircuits remain unknown. Here, we show that acetylcholine release from cholinergic septohippocampal projections causes a long-lasting GABAergic inhibition of hippocampal dentate granule cells in vivo and in vitro. This inhibition is caused by cholinergic activation of hilar astrocytes, which provide glutamatergic excitation of hilar inhibitory interneurons. These results demonstrate that acetylcholine release can cause slow inhibition of principal neuronal activity via astrocyte intermediaries.

Function and developmental origin of a mesocortical inhibitory circuit.

Midbrain ventral tegmental neurons project to the prefrontal cortex and modulate cognitive functions. Using viral tracing, optogenetics and electrophysiology, we found that mesocortical neurons in the mouse ventrotegmental area provide fast glutamatergic excitation of GABAergic interneurons in the prefrontal cortex and inhibit prefrontal cortical pyramidal neurons in a robust and reliable manner. These mesocortical neurons were derived from a subset of dopaminergic progenitors, which were dependent on prolonged Sonic Hedgehog signaling for their induction. Loss of these progenitors resulted in the loss of the mesocortical inhibitory circuit and an increase in perseverative behavior, whereas mesolimbic and mesostriatal dopaminergic projections, as well as impulsivity and attentional function, were largely spared. Thus, we identified a previously uncharacterized mesocortical circuit contributing to perseverative behaviors and found that the diversity of dopaminergic neurons begins to be established during their progenitor phase.

Activity of fin muscles and fin motoneurons during swimming motor pattern in the lamprey.

Coordination of motoneuron activity is a fundamental prerequisite for the generation of functional locomotor patterns. We investigate the neural mechanisms that coordinate activity of motoneuron pools in the vertebrate spinal cord with differing phases of activity in the locomotor cycle in a simple motor system, the lamprey swimming network. In the region of dorsal fins the lamprey spinal cord contains two groups of motoneurons: the myotomal motoneurons that innervate the trunk muscles; and the fin motoneurons controlling muscle fibres of the dorsal fins. We investigated the activity of fin muscles during swimming in vivo and that of fin motoneurons during fictive swimming in vitro. During swimming in vivo with cycle periods of 4-8 Hz, fin muscle activity covered a broad portion of the cycle, with the peak of activity out-of-phase to the ipsilateral myotomal muscles. During fictive swimming evoked by N-methyl-d-aspartate in the isolated spinal cord, fin motoneurons expressed similar out-of-phase activity. The phase relationship of the synaptic drive to fin motoneurons was examined by recording their activity intracellular during fictive swimming. Three different forms of membrane potential oscillation with different time courses in the locomotor cycle could be distinguished. Sagittal lesions of the spinal cord in the segment where fin motoneurons are recorded and up to one segment rostral and caudal from it did not influence the out-of-phase activity pattern of the motoneurons. Our results indicate that coordination of fin motoneuron activity with the locomotor activity of myotomal motoneurons does not depend on intrasegmental contralateral premotor elements.