Individual cortical neurons can selectively respond to specific environmental features, such as visual motion or faces. How this relates to the selectivity of the presynaptic network across cortical layers remains unclear. We used single-cell-initiated, monosynaptically restricted retrograde transsynaptic tracing with rabies viruses expressing GCaMP6s to image, in vivo, the visual motion-evoked activity of individual layer 2/3 pyramidal neurons and their presynaptic networks across layers in mouse primary visual cortex. Neurons within each layer exhibited similar motion direction preferences, forming layer-specific functional modules. In one-third of the networks, the layer modules were locked to the direction preference of the postsynaptic neuron, whereas for other networks the direction preference varied by layer. Thus, there exist feature-locked and feature-variant cortical networks.

We study the integration of multisensory and central input at the level of an identified fly motoneuron, the ventral cervical nerve motoneuron (VCNM) cell, which controls head movements of the animal. We show that this neuron receives input from a central neuron signaling flight activity, from two identified wide-field motion-sensitive neurons, from the wind-sensitive Johnston organ on the antennae, and from the campaniform sensillae of the halteres. We find that visual motion alone leads to only subthreshold responses. Only when it is combined with flight activity or wind stimuli does the VCNM respond to visual motion by modulating its spike activity in a directionally selective way. This nonlinear enhancement of visual responsiveness in the VCNM by central activity is reflected at the behavioral level, when compensatory head movements are measured in response to visual motion. While head movements of flies have only a small amplitude when flies are at rest, the response amplitude is increased by a factor of 30-40 during flight.behavioral state | electrophysiology | motion vision | proprioceptor T he perceptual response to a sensory stimulus depends on the behavioral state of the animal. This is especially evident when responses between awake and sleeping or anesthetized animals are compared (1-5). In addition, attention can strongly modulate the perceptual response (6-11). Furthermore, recent studies have shown that the response properties of sensory neurons depend on the locomotor state of the animal (12-16). However, it is not clear at what level of the nervous system the integration of sensory input and internal state occurs and according to which rules. We address this question in an identified ventral cervical nerve motoneuron (VCNM) that is part of the neck motor system of the blowfly Calliphora vicina.Blowflies are well known for their aerobatic maneuvers. This flight behavior has important consequences for vision, because fast turns can lead to motion blur and thus impair vision of spatial details. These adverse effects on vision can, in principle, be partly compensated for by eye movements. However, in all insects, the eyes are fixed to the head capsule and only small eye movements are possible (17). Therefore, effective compensatory eye movements can only be made by turning the head. Blowflies can turn their head relative to the thorax around all three body axes but with different amplitudes (yaw, ±20°; pitch, ±20°; and roll, ±90°). Accordingly, movements of the thorax during flight are compensated for by countermovements of the head relative to the thorax (18-21). These head movements are mediated by the neck motor system, which consists of 21 pairs of muscles, with each of them innervated by a single motoneuron (22,23). The three oblique horizontal muscles, OH3-5, are supplied by the ventral cervical nerve, comprising the axons of three identical motor neurons, . Unilateral contraction of these muscles leads to a sidewise deflection of the head (yaw response), whereas bilateral contraction pulls th...

Many motion-sensitive tangential cells of the lobula plate in blowflies are well described with respect to their visual response properties and the connectivity among them. They have large and complex receptive fields with different preferred directions in different parts of their receptive fields matching the optic flow that occurs during various flight maneuvers. However, much less is known about how tangential cells connect to postsynaptic neurons descending to the motor circuits in the thoracic ganglion and how optic flow is represented in these downstream neurons. Here we describe the physiology and the connectivity of a prominent descending neuron called DNOVS1 (for descending neurons of the ocellar and vertical system). We find that DNOVS1 is electrically coupled to a subset of vertical system cells. The specific wiring leads to a preference of DNOVS1 for rotational flow fields around a particular body axis. In addition, DNOVS1 receives input from interneurons connected to the ocelli.

For visual orientation and course stabilization, flies rely heavily on the optic flow perceived by the animal during flight. The processing of optic flow is performed in motion-sensitive tangential cells of the lobula plate, which are well described with respect to their visual response properties and the connectivity among them. However, little is known about the postsynaptic descending neurons, which convey motion information to the motor circuits in the thoracic ganglion. Here we investigate the physiology and connectivity of an identified premotor descending neuron, called DNOVS2 (for descending neuron of the ocellar and vertical system). We find that DNOVS2 is tuned in a supralinear way to rotation around the longitudinal body axis. Experiments involving stimulation of the ipsilateral and the contralateral eye indicate that ipsilateral computation of motion information is modified nonlinearly by motion information from the contralateral eye. Performing double recordings of DNOVS2 and lobula plate tangential cells, we find that DNOVS2 is connected ipsilaterally to a subset of vertical-sensitive cells. From the contralateral eye, DNOVS2 receives input most likely from V2, a heterolateral spiking neuron. This specific neural circuit is sufficient for the tuning of DNOVS2, making it probably an important element in optomotor roll movements of the head and body around the fly's longitudinal axis.

For a moving animal, optic flow is an important source of information about its ego-motion. In flies, the processing of optic flow is performed by motion sensitive tangential cells in the lobula plate. Amongst them, cells of the vertical system (VS cells) have receptive fields with similarities to optic flows generated during rotations around different body axes. Their output signals are further processed by pre-motor descending neurons. Here, we investigate the local motion preferences of two descending neurons called descending neurons of the ocellar and vertical system (DNOVS1 and DNOVS2). Using an LED arena subtending 240°9 95°of visual space, we mapped the receptive fields of DNOVS1 and DNOVS2 as well as those of their presynaptic elements, i.e. VS cells 1-10 and V2. The receptive field of DNOVS1 can be predicted in detail from the receptive fields of those VS cells that are most strongly coupled to the cell. The receptive field of DNOVS2 is a combination of V2 and VS cells receptive fields. Predicting the global motion preferences from the receptive field revealed a linear spatial integration in DNOVS1 and a superlinear spatial integration in DNOVS2. In addition, the superlinear integration of V2 output is necessary for DNOVS2 to differentiate between a roll rotation and a lift translation of the fly.

In many species, motion-sensitive neurons responding to optic flow at higher processing stages are well characterized; however, less is known how this representation of ego-motion is further transformed into an appropriate motor response. Here, we analyzed in the blowfly Calliphora vicina the visuomotor transformation from motion-sensitive neurons in the lobula plate [V2 and vertical system (

Genetic engineering by viral infection of single cells is useful to study complex systems such as the brain. However, available methods for infecting single cells have drawbacks that limit their applications. Here we describe 'virus stamping', in which viruses are reversibly bound to a delivery vehicle-a functionalized glass pipette tip or magnetic nanoparticles in a pipette-that is brought into physical contact with the target cell on a surface or in tissue, using mechanical or magnetic forces. Different single cells in the same tissue can be infected with different viruses and an individual cell can be simultaneously infected with different viruses. We use rabies, lenti, herpes simplex, and adeno-associated viruses to drive expression of fluorescent markers or a calcium indicator in target cells in cell culture, mouse retina, human brain organoid, and the brains of live mice. Virus stamping provides a versatile solution for targeted single-cell infection of diverse cell types, both in vitro and in vivo.

For sea slugs, chemosensory information represents an important sensory modality, because optical and acoustical information are limited. In the present study, we focussed on the neuroanatomy of the rhinophores and processing of olfactory stimuli in the rhinophore ganglion of Archidoris pseudoargus, belonging to the order of Nudibranchia in the subclass of Opisthobranchia. Histological techniques, Xuorescent markers, and immunohistochemistry were used to analyse neuroanatomical features of the rhinophore. A large ganglion and a prominent central lymphatic channel are surrounded by longitudinal muscles. Many serotonin-immunoreactive (IR) processes were found around the centre and between the ganglion and the highly folded lobes of the rhinophore, but serotonin-IR cell bodies were absent inside the rhinophore. In contrast to the conditions recently found in Aplysia punctata, we found no evidence for the presence of olfactory glomeruli within the rhinophore. Using calcium-imaging techniques with Fura II as a calcium indicator, we found diVerential calcium responses in various regions within the ganglion to stimulation of the rhinophore with diVerent amino acids. The lack of glomeruli in the rhinophores induces functional questions about processing of chemical information in the rhinophore.