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Behavioral flexibility and timely reactions to salient stimuli are essential for survival. The subcortical thalamic-basolateral amygdala (BLA) pathway serves as a shortcut for salient stimuli ensuring rapid processing. Here, we show that BLA neuronal and thalamic axonal activity mirror the defensive behavior evoked by an innate visual threat as well as an auditory learned threat. Importantly, perturbing this pathway compromises defensive responses to both forms of threats, in that animals fail to switch from exploratory to defensive behavior. Despite the shared pathway between the two forms of threat processing, we observed noticeable differences. Blocking beta-adrenergic receptors impair the defensive response to the innate but not the learned threats. This reduced defensive response, surprisingly, is reflected in the suppression of the activity exclusively in the BLA, as the thalamic input response remains intact. Our side-by-side examination highlights the similarities and differences between innate and learned threat-processing, thus providing new fundamental insights.

The asymmetric summation of kinetically distinct glutamate inputs across the dendrites of retinal ‘starburst’ amacrine cells is one of the several mechanisms that have been proposed to underlie their direction-selective properties, but experimentally verifying input kinetics has been a challenge. Here, we used two-photon glutamate sensor (iGluSnFR) imaging to directly measure the input kinetics across individual starburst dendrites. We found that signals measured from proximal dendrites were relatively sustained compared to those measured from distal dendrites. These differences were observed across a range of stimulus sizes and appeared to be shaped mainly by excitatory rather than inhibitory network interactions. Temporal deconvolution analysis suggests that the steady-state vesicle release rate was ~3 times larger at proximal sites compared to distal sites. Using a connectomics-inspired computational model, we demonstrate that input kinetics play an important role in shaping direction selectivity at low stimulus velocities. Taken together, these results provide direct support for the ‘space-time wiring’ model for direction selectivity.

The long-term administration of tamoxifen to estrogen receptor α (ERα)-positive breast cancer patients is an established treatment that reduces mortality and recurrence. However, resistance to tamoxifen and an increased risk of endometrial cancer may occur; therefore, the mechanisms by which tamoxifen causes these adverse effects warrant further study. Tamoxifen has been shown to activate mitogen-activated protein kinase (MAPK) in an ERα-independent manner; therefore, we investigated its effects on the MAPK-mediated non-canonical activation of EphA2, a critical event regulating cell migration. Tamoxifen at slightly higher concentrations induced the rapid phosphorylation of EphA2 at Ser-897 via the MAPK/extracellular signal-regulated kinase (ERK) kinase (MEK)-ERK-ribosomal S6 kinases (RSK) pathway in HeLa cells. In addition, tamoxifen significantly enhanced the migration ability of ERα-negative MDA-MB-231 breast cancer cells in RSK- and EphA2-dependent manners. Phosphorylated EphA2 was internalized and re-localized to the plasma membrane, including lamellipodia, in an RSK-dependent manner. Collectively, the present results provide novel insights into the tumor-promoting activity of tamoxifen.

A Correction to this paper has been published: https://doi.org/10.1038/s41467-021-22763-3

Summary Locomotion creates various patterns of optic flow on the retina, which provide the observer with information about their movement relative to the environment. However, it is unclear how these optic flow patterns are encoded by the cortex. Here, we use two-photon calcium imaging in awake mice to systematically map monocular and binocular responses to horizontal motion in four areas of the visual cortex. We find that neurons selective to translational or rotational optic flow are abundant in higher visual areas, whereas neurons suppressed by binocular motion are more common in the primary visual cortex. Disruption of retinal direction selectivity in Frmd7 mutant mice reduces the number of translation-selective neurons in the primary visual cortex and translation- and rotation-selective neurons as well as binocular direction-selective neurons in the rostrolateral and anterior visual cortex, blurring the functional distinction between primary and higher visual areas. Thus, optic flow representations in specific areas of the visual cortex rely on binocular integration of motion information from the retina., Highlights • Translation- and rotation-selective neurons are abundant in higher visual areas • Optic-flow-selective neurons in V1 and RL/A rely on retinal direction selectivity • Retinal direction selectivity controls functional segregation between V1 and RL/A • Binocular integration of retinal motion information underlies optic flow selectivity, Locomotion creates different patterns of optic flow, but how these are encoded by the cortex is unclear. By mapping cortical activity in awake mice, Rasmussen et al. demonstrate that binocular integration of retinal motion direction selectivity causally influences optic flow selectivity of neurons residing in distinct areas of the visual cortex.

The ability to encode the direction of image motion is fundamental to our sense of vision. Direction selectivity along the four cardinal directions is thought to originate in direction-selective ganglion cells (DSGCs), due to directionally-tuned GABAergic suppression by starburst cells. Here, by utilizing two-photon glutamate imaging to measure synaptic release, we reveal that direction selectivity along all four directions arises earlier than expected, at bipolar cell outputs. Thus, DSGCs receive directionally-aligned glutamatergic inputs from bipolar cell boutons. We further show that this bouton-specific tuning relies on cholinergic excitation and GABAergic inhibition from starburst cells. In this way, starburst cells are able to refine directional tuning in the excitatory visual pathway by modulating the activity of DSGC dendrites and their axonal inputs using two different neurotransmitters.

The brain monitors the sensory environment via signals from the sensory periphery, such as the olfactory epithelium, the inner ear, and the retina. Understanding how sensory stimuli are processed throughout the sensory hierarchy, and how this relates to behavior, is a central outstanding question in the field of neuroscience. The processing of visual motion in mice offers unique opportunities for addressing these questions thanks to a rich literature on the anatomical and physiological properties of motion-sensitive neurons across the visual system, paired with recent developments of cutting-edge genetic and imaging approaches. A visual scene typically contains motion originating from either moving objects or optic flow caused by self-generated movements. Neurons encoding the direction of visual motion are said to be ‘direction-selective’. It was historically believed the circuits responsible for creating direction selectivity de novo exist within the visual cortex. Yet, in mice, direction-selective responses can be found already in the retina, suggesting in this model organism visual motion analysis starts at the earliest stage of the visual hierarchy. This minireview presents emerging literature demonstrating how retinal direction-selective cells make causal contributions to central visual motion processing and visually guided behaviors in mice, and their potential clinical relevance, and outlines experiments for testing remaining questions. Research in this field will undoubtedly continue to advance our understanding of the basic principles of the visual system and how sensory neurons extract fundamental features of the world.

Eye-tracking is a method for tracking the position of the eye and size of the pupil, often employed in neuroscience laboratories and clinics. Eye-trackers are widely used, from studying brain dynamics to investigating neuropathology and disease models. Despite this broad utility, eye-trackers are expensive, hardware-intensive, and proprietary, which have limited this approach to high-resource facilities. Besides, experiments have largely been confined to static open-loop designs and post hoc analysis due to the inflexibility of current systems. Here, we developed an open-source eye-tracking system, named EyeLoop, tailored to dynamic experiments. This Python-based software easily integrates custom functions via a modular logic, tracks a multitude of eyes, including rodent, human, and non-human primate eyes, and it operates well on inexpensive consumer-grade hardware. One of the most appealing applications of EyeLoop is closed-loop experiments, in which the eyes evoke stimulus feedback, such as rapid neuronal optogenetic stimulation. By using EyeLoop, we demonstrate its utility in an open-loop, a closed-loop, and a biomedical experiment. With a remarkably low minimal hardware cost amounting to 29 USD, EyeLoop makes dynamic eye-tracking accessible to low-resource facilities, such as high schools, small laboratories, and small clinics.

In many parts of the central nervous system, including the retina, it is unclear whether cholinergic transmission is mediated by rapid, point-to-point synaptic mechanisms, or slower, broad-scale ‘non-synaptic’ mechanisms. Here, we characterized the ultrastructural features of cholinergic connections between direction-selective starburst amacrine cells and downstream ganglion cells in an existing serial electron microscopy data set, as well as their functional properties using electrophysiology and two-photon acetylcholine (ACh) imaging. Correlative results demonstrate that a ‘tripartite’ structure facilitates a ‘multi-directed’ form of transmission, in which ACh released from a single vesicle rapidly (~1 ms) co-activates receptors expressed in multiple neurons located within ~1 µm of the release site. Cholinergic signals are direction-selective at a local, but not global scale, and facilitate the transfer of information from starburst to ganglion cell dendrites. These results suggest a distinct operational framework for cholinergic signaling that bears the hallmarks of synaptic and non-synaptic forms of transmission., Cholinergic neurons may transmit information via fast synaptic, point-to-point signaling or diffuse, slow extra-synaptic signaling. The authors show that ACh from a single vesicle triggers synchronous miniature currents in two neurons, showing that ACh can spread significant distances to drive rapid ‘synaptic’ signals.

Acetylcholine (ACh) is a key neurotransmitter that plays diverse roles in many parts of the central nervous system, including the retina. However, assessing the precise spatiotemporal dynamics of ACh is technically challenging and whether ACh transmits signals via rapid, point-to-point synaptic mechanisms, or broader-scale ‘non-synaptic’ mechanisms has been difficult to ascertain. Here, we examined the properties of cholinergic transmission at individual contacts made between direction-selective starburst amacrine cells and downstream ganglion cells in the retina. Using a combination of electrophysiology, serial block-face electron microscopy, and two-photon ACh imaging, we demonstrate that ACh signaling bears the hallmarks of both non-synaptic and synaptic forms of transmission. ACh co-activates nicotinic ACh receptors located on the intersecting dendrites of pairs of ganglion cells, with equal efficiency (non-synaptic)— and yet retains the ability to generate rapid ‘miniature’ currents (∼1 ms rise times: synaptic). Fast cholinergic signals do not appear to depend on anatomically well-defined synaptic structures. We estimate that ACh spread is limited to ∼1-2 µm from its sites of release, which may help starbursts drive local direction-selective cholinergic responses in ganglion cell dendrites. Together, our results establish the functional architecture for cholinergic signaling at a central synapse and propose a novel motif whereby single presynaptic sites can co-transmit information to multiple neurons on a millisecond timescale.

Visual features extracted by retinal circuits are streamed into higher visual areas (HVAs) after being processed along the visual hierarchy. However, how specialized neuronal representations of HVAs are built, based on retinal output channels, remained unclear. Here, we addressed this question by determining the effects of genetically disrupting retinal direction selectivity on motion-evoked responses in visual stages from the retina to HVAs in mice. Direction-selective (DS) cells in the rostrolateral (RL) area that prefer higher temporal frequencies, and that change direction tuning bias as the temporal frequency of a stimulus increases, are selectively reduced upon retinal manipulation. DS cells in the primary visual cortex projecting to area RL, but not to the posteromedial area, were similarly affected. Therefore, the specific connectivity of cortico-cortical projection neurons routes feedforward signaling originating from retinal DS cells preferentially to area RL. We thus identify a cortical processing stream for motion computed in the retina., Visual features are streamed into higher visual areas (HVAs), but how representations in HVAs are built, based on retinal output channels, is unknown. Here, the authors show that specific connectivity of cortical neurons routes retina-originated direction-selective signaling into distinct HVAs.

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.

Summary The ability to encode the direction of image motion is fundamental to our sense of vision. Direction selectivity along the four cardinal directions is thought to originate in direction-selective ganglion cells (DSGCs) because of directionally tuned GABAergic suppression by starburst cells. Here, by utilizing two-photon glutamate imaging to measure synaptic release, we reveal that direction selectivity along all four directions arises earlier than expected at bipolar cell outputs. Individual bipolar cells contained four distinct populations of axon terminal boutons with different preferred directions. We further show that this bouton-specific tuning relies on cholinergic excitation from starburst cells and GABAergic inhibition from wide-field amacrine cells. DSGCs received both tuned directionally aligned inputs and untuned inputs from among heterogeneously tuned glutamatergic bouton populations. Thus, directional tuning in the excitatory visual pathway is incrementally refined at the bipolar cell axon terminals and their recipient DSGC dendrites by two different neurotransmitters co-released from starburst cells., Graphical abstract, Highlights • Cardinal direction selectivity emerges at types 7 and 2 bipolar cell axon terminals • Starburst amacrine cells are necessary for direction selectivity in bipolar cells • Cholinergic excitation and GABAergic inhibition are integrated at axon terminals • Direction-selective ganglion cells receive directionally aligned glutamate inputs, Matsumoto et al. uncover that cardinal direction selectivity emerges in retinal bipolar cell axon terminals. The selectivity is established de novo at the terminals by starburst amacrine cells, mediated by both cholinergic transmission and wide-field amacrine cells. The bipolar cells transmit tuned glutamate inputs to directional preference-matched ganglion cells.

The detection of visual motion is a fundamental function of the visual system. How motion speed and direction are computed together at the cellular level, however, remains largely unknown. Here, we suggest a circuit mechanism by which excitatory inputs to direction-selective ganglion cells in the mouse retina become sensitive to the motion speed and direction of image motion. Electrophysiological, imaging, and connectomic analyses provide evidence that the dendrites of ON direction-selective cells receive spatially offset and asymmetrically filtered glutamatergic inputs along motion-preference axis from asymmetrically wired bipolar and amacrine cell types with distinct release dynamics. A computational model shows that, with this spatiotemporal structure, the input amplitude becomes sensitive to speed and direction by a preferred direction enhancement mechanism. Our results highlight the role of an excitatory mechanism in retinal motion computation by which feature selectivity emerges from non-selective inputs. Copyright © 2019 The Author(s). Published by Elsevier Ltd.. All rights reserved.

ispartof: FRONTIERS IN NEURAL CIRCUITS vol:13 ispartof: location:Switzerland status: published

Learning drives behavioral adaptations necessary for survival. While plasticity of excitatory projection neurons during associative learning has been extensively studied, little is known about the contributions of local interneurons. Using fear conditioning as a model for associative learning, we found that behaviorally relevant, salient stimuli cause learning by tapping into a local microcircuit consisting of precisely connected subtypes of inhibitory interneurons. By employing deep-brain calcium imaging and optogenetics, we demonstrate that vasoactive intestinal peptide (VIP)-expressing interneurons in the basolateral amygdala are activated by aversive events and provide a mandatory disinhibitory signal for associative learning. Notably, VIP interneuron responses during learning are strongly modulated by expectations. Our findings indicate that VIP interneurons are a central component of a dynamic circuit motif that mediates adaptive disinhibitory gating to specifically learn about unexpected, salient events, thereby ensuring appropriate behavioral adaptations.

Learning drives behavioral adaptations necessary for survival. While plasticity of excitatory projection neurons during associative learning is studied extensively, little is known about the contributions of local interneurons. Using fear conditioning as a model for associative learning, we find that behaviorally relevant, salient stimuli cause learning by tapping into a local microcircuit consisting of precisely connected subtypes of inhibitory interneurons. By employing calcium imaging and optogenetics, we demonstrate that vasoactive intestinal peptide (VIP)-expressing interneurons in the basolateral amygdala are activated by aversive events and provide an instructive disinhibitory signal for associative learning. Notably, VIP interneuron responses are plastic and shift from the instructive to the predictive cue upon memory formation. We describe a novel form of adaptive disinhibitory gating by VIP interneurons that allows to discriminate unexpected, important from irrelevant information, and might be a general dynamic circuit motif to trigger stimulus-specific learning, thereby ensuring appropriate behavioral adaptations to salient events.

Each mosaic of retinal ganglion cells is thought to extract the same visual feature across mouse retina, but a new study shows that ganglion cells of the same type actually show different light response depending on retinal location. Each mosaic of retinal ganglion cells is thought to extract the same visual feature across mouse retina. A new study shows that ganglion cells of the same type actually show different light response depending on retinal location.

Tracing cells that project to one neuron Feature extraction is a prominent characteristic of cortical neurons involved in the early stages of sensory processing. Wertz et al. retrogradely marked an injected neuron and its direct inputs to reveal the network mechanisms that mediate their response. Neurons within each presynaptic network layer of single direction-selective cells showed similar motion direction preferences. In some networks, layer-specific functional modules were identical to the orientation preference of the postsynaptic neuron. Presynaptic neurons, however, displayed a general bias toward the stimulus feature that elicited a response in the postsynaptic neuron. Science , this issue p. 70

Summary The thalamus receives sensory input from different circuits in the periphery. How these sensory channels are integrated at the level of single thalamic cells is not well understood. We performed targeted single-cell-initiated transsynaptic tracing to label the retinal ganglion cells that provide input to individual principal cells in the mouse lateral geniculate nucleus (LGN). We identified three modes of sensory integration by single LGN cells. In the first, 1–5 ganglion cells of mostly the same type converged from one eye, indicating a relay mode. In the second, 6–36 ganglion cells of different types converged from one eye, revealing a combination mode. In the third, up to 91 ganglion cells converged from both eyes, revealing a binocular combination mode in which functionally specialized ipsilateral inputs joined broadly distributed contralateral inputs. Thus, the LGN employs at least three modes of visual input integration, each exhibiting different degrees of specialization., Highlights • Individual LGN cells integrate retinal inputs in one of three distinct modes • Relay-mode cells integrate inputs from few retinal ganglion cells of mostly one type • Combination- and binocular-mode cells combine inputs from many ganglion cell types • The three integration modes exhibit different degrees of cell-type specialization, Rompani et al. employ single-cell-initiated transsynaptic tracing to decipher patterns of input integration in the thalamus. They show that individual cells in the lateral geniculate nucleus integrate retinal inputs in three distinct modes, each exhibiting different degrees of specialization.

Kebschull et al. (2016a) describe “MAPseq,” which tags individual neurons from a specific brain region with individual mRNA barcodes and sequences these barcodes in other brain regions. This allows the simultaneous mapping of long-range neuronal projections at single-cell resolution.

Summary Neuronal circuit asymmetries are important components of brain circuits, but the molecular pathways leading to their establishment remain unknown. Here we found that the mutation of FRMD7, a gene that is defective in human congenital nystagmus, leads to the selective loss of the horizontal optokinetic reflex in mice, as it does in humans. This is accompanied by the selective loss of horizontal direction selectivity in retinal ganglion cells and the transition from asymmetric to symmetric inhibitory input to horizontal direction-selective ganglion cells. In wild-type retinas, we found FRMD7 specifically expressed in starburst amacrine cells, the interneuron type that provides asymmetric inhibition to direction-selective retinal ganglion cells. This work identifies FRMD7 as a key regulator in establishing a neuronal circuit asymmetry, and it suggests the involvement of a specific inhibitory neuron type in the pathophysiology of a neurological disease. Video Abstract, Highlights • FRMD7 is required for the horizontal optokinetic reflex in mice as in humans • Horizontal direction selectivity is lost in the retina of FRMD7 mutant mice • Asymmetry of inhibitory inputs to horizontal DS cells is lost in FRMD7 mutant mice • FRMD7 is expressed in ChAT-expressing cells in the retina of mice and primates, Yonehara et al. show that FRMD7, a gene that is defective in human congenital nystagmus, is required in the mouse retina to establish spatially asymmetric inhibitory inputs from starburst cells to horizontal direction-selective ganglion cells.

Adeno-associated viruses (AAVs) are commonly used for in vivo gene transfer. Nevertheless, AAVs that provide efficient transduction across specific organs or cell populations are needed. Here, we describe AAV-PHP.eB and AAV-PHP.S, capsids that efficiently transduce the central and peripheral nervous systems, respectively. In the adult mouse, intravenous administration of 1×1011 vector genomes (vg) of AAV-PHP.eB transduced 69% of cortical and 55% of striatal neurons, while 1×1012 vg AAV-PHP.S transduced 82% of dorsal root ganglion neurons, as well as cardiac and enteric neurons. The efficiency of these vectors facilitates robust co-transduction and stochastic, multicolor labeling for individual cell morphology studies. To support such efforts, we provide methods for labeling a tunable fraction of cells without compromising color diversity. Furthermore, when used with cell type-specific promoters, these AAVs provide targeted gene expression across the nervous system and enable efficient and versatile gene manipulation throughout the nervous system of transgenic and non-transgenic animals.

The ability to detect motion in the visual scene is a fundamental computation in the visual system that is first performed in the retina. The cells responsible for encoding motion direction are the direction-sensitive ganglion cells (DSGCs), which fire a maximum number of action potentials during movement in one direction and fire minimally during movement in the opposite direction. Highly selective wiring from inhibitory cells contributes to determining the direction-selection characteristics of these ganglion cells, yet how the asymmetric wiring inherent to these connections is established was unknown. Two groups using complementary techniques, including pharmacology, electrophysiology and optogenetics, report that although inhibitory inputs to both sides of the direction-selective cell are uniform early in development, by the second postnatal week, inhibitory synapses on the null side strengthen while those on the preferred side remain constant. These plasticity changes occur independent of neural activity, suggesting a specific developmental program is executed to produce the direction-selective circuitry in the retina. In the retina, highly selective wiring from inhibitory cells contributes to determine the direction-selection characteristics of an individual ganglion cell, yet how the asymmetric wiring inherent to these connections is established was unknown. Here, two independent studies using complementary techniques, including pharmacology, electrophysiology and optogenetics, find that although inhibitory inputs to both sides of the direction-selective cell are uniform early in development, by the second postnatal week, inhibitory synapses on the null side strengthen whereas those on the preferred side remain constant. These plasticity changes occur independent of neural activity, indicating that a specific developmental program is executed to produce the direction-selective circuitry in the retina. Spatial asymmetries in neural connectivity have an important role in creating basic building blocks of neuronal processing1,2. A key circuit module of directionally selective (DS) retinal ganglion cells is a spatially asymmetric inhibitory input from starburst amacrine cells3,4,5. It is not known how and when this circuit asymmetry is established during development. Here we photostimulate mouse starburst cells targeted with channelrhodopsin-2 (refs 6–8) while recording from a single genetically labelled type of DS cell9,10. We follow the spatial distribution of synaptic strengths between starburst and DS cells during early postnatal development before these neurons can respond to a physiological light stimulus, and confirm connectivity by monosynaptically restricted trans-synaptic rabies viral tracing. We show that asymmetry develops rapidly over a 2-day period through an intermediate state in which random or symmetric synaptic connections have been established. The development of asymmetry involves the spatially selective reorganization of inhibitory synaptic inputs. Intriguingly, the spatial distribution of excitatory synaptic inputs from starburst cells is significantly more symmetric than that of the inhibitory inputs at the end of this developmental period. Our work demonstrates a rapid developmental switch from a symmetric to asymmetric input distribution for inhibition in the neural circuit of a principal cell.

SummaryA new study using zebrafish genetics and whole-brain imaging has identified more than 50 retinal ganglion cell morphologies and produced the first comprehensive map of connectivity between retina and its target visual centers.

We previously identified SPARC-related protein-containing immunoglobulin domains 1 (SPIG1, also known as Follistatin-like protein 4) as one of the dorsal-retina-specific molecules expressed in the developing chick retina. We here demonstrated that the knockdown ofSPIG1in the retinal ganglion cells (RGCs) of developing chick embryos induced the robust ectopic branching of dorsal RGC axons and failed to form a tight terminal zone at the proper position on the tectum. The knockdown ofSPIG1in RGCs also led to enhanced axon branchingin vitro. However, this was canceled by the addition of a neutralizing antibody against brain-derived neurotrophic factor (BDNF) to the culture medium. SPIG1 and BDNF were colocalized in vesicle-like structures in cells. SPIG1 bound with the proform of BDNF (proBDNF) but very weakly with mature BDNFin vitro. The expression and secretion of mature BDNF were significantly decreased when SPIG1 was exogenously expressed with BDNF in HEK293T or PC12 cells. The amount of mature BDNF proteins as well as the tyrosine phosphorylation level of the BDNF receptor, tropomyosin-related kinase B (TrkB), in the hippocampus were significantly higher inSPIG1-knockout mice than in wild-type mice. Here the spine density of CA1 pyramidal neurons was consistently increased. Together, these results suggest that SPIG1 negatively regulated BDNF maturation by binding to proBDNF, thereby suppressing axonal branching and spine formation.

Destination Specificity During brain development, some types of neurons migrate from where they are born to their final functional locations. Some neurons migrate radially, from the inside to the outside, while others migrate tangentially. Di Meglio et al. (p. 204 ) analyzed the migration of a group of tangentially migrating neurons in the hindbrain. Although these neurons all entered the same migratory stream, they each retained positional information such that their relative organization in the destination site reflected their original organization. Interactions between epigenetic signals and the genes encoding Hox transcription factors encoded the positional information and fine-tuned migration.