Optic Tectum Research Articles

Insecticide-induced changes in amphibian brains: How sublethal concentrations of chlorpyrifos directly affect neurodevelopment.

Widespread use of pesticides often contaminates natural habitats, exposing nontarget organisms to pesticides that were designed to control pest populations. Even low levels of pesticides can affect aquatic communities both directly and indirectly. Previous work has shown that trace amounts of the pesticide chlorpyrifos altered tadpole morphology and neurodevelopment in artificial ponds (mesocosms). To determine whether effects resulted from direct chlorpyrifos exposure or from disruption of the food web due to a pesticide-induced decline in zooplankton, we examined the impacts of chlorpyrifos on amphibian development in the presence of chlorpyrifos-resistant zooplankton, a key component of the aquatic trophic community. Northern leopard frog (Lithobates pipiens) tadpoles were reared through metamorphosis in mesocosms containing either 0 or 1 µg/L chlorpyrifos and either chlorpyrifos-resistant or chlorpyrifos-sensitive Daphnia pulex zooplankton. Developmental exposure to chlorpyrifos resulted in metamorphs with a relatively wider optic tectum, medulla, and diencephalon compared with controls, and this result was found regardless of the zooplankton population within the mesocosm. Thus, chlorpyrifos directly impacted brain development, independent of the effects on the trophic community. With respect to body shape, chlorpyrifos had no effect on body shape of metamorphs reared in mesocosms with chlorpyrifos-sensitive zooplankton, but body shape was sensitive to zooplankton population in the absence of chlorpyrifos. To conclude, low, ecologically relevant doses of organophosphorous pesticides can directly impact neurodevelopment in a vertebrate model. Environ Toxicol Chem 2018;37:2692-2698. © 2018 SETAC.

Superior Colliculus: A Vision for Orienting

The superior colliculus contains neurons sensitive to motion direction. New research shows that these neurons are anatomically clustered: those representing the region of visual space 'seen' by both eyes preferentially respond to nasal motion directions and others to opposite directions.

The Pax2 and Pax6 Transcription Factors in the Optic Nerve and Brain of Trout Oncorhynchus mykiss after a Mechanical Eye Injury

The distribution of the Pax2+ transcription factor in the optic nerve after a mechanical eye injury on the side of damage and in the contralateral nerve has been studied in the trout Oncorhynchus mykiss. It has been found that injury of the optic nerve in this fish species causes Pax2+ reactive astrocytes involved in the initial stages of optic nerve axon regeneration to increase in number, especially in the area of the head and the proximal part of the optic nerve. As the optic nerve in trout is damaged, a significant growth of the heterogeneous population of Pax6+ cells occurs in the brain divisions that have direct retinal inputs, diencephalon, and optic tectum. A part of the Pax6+ cells have an undifferentiated phenotype and are a component of reactive neurogenic niches located in the periventricular zone and parenchymal regions of the brain. Another population of Pax6+ cells has the radial glial phenotype and appears as a result of activation of the constitutive neurogenic domains also within the newly formed reactive neurogenic niches. Thus, due to the optic nerve injury, a pronounced neurogenic response associated with the appearance of reactive neurogenic niches and radial glia arises both in the brain divisions with direct retinal projections and in those lacking the retinal projections as well as in remote regions. The results obtained indicate that the damage to the optic nerve causes an increased reactive neurogenesis in the brain of adult trout.

NGF, TrkA-P and neuroprotection after a hypoxic event in the developing central nervous system

A decrease in the concentration of oxygen in the blood and tissues (hypoxia) produces important, sometimes irreversible, damages in the central nervous system (CNS) both during development and also postnatally. The present work aims at analyzing the expression of nerve growth factor (NGF) and p75 and the activation of TrkA in response to an acute normobaric hypoxic event and to evaluate the possible protective role of exogenous NGF. The developing chick optic tectum (OT), a recognized model of corticogenesis, was used as experimental system by means of in vivo and in vitro studies.Based on identification of the period of highest sensitivity of developmental programmed cell death (ED15) we show that hypoxia has a mild but reproducible effect that consist of a temporal increase of cell death 6 h after the end of a hypoxic treatment. Cell death was preceded by a significant early increase in the expression of Nerve Growth Factor (NGF) and its membrane receptor p75. In addition, we found a biphasic response of TrkA activation: a decrease during hypoxia followed by an increase −4 h later- that temporally coincide with the interval of NGF overexpression. To test the NGF - NGF receptors role in hypoxic cell death, we quantified, in primary neuronal cultures derived from ED15 OT, the levels of TrkA activation after an acute hypoxic treatment. A significant decline in the level of TrkA activation was observed during hypoxia followed, 24 h later, by significant cell death. Interestingly, this cell death can be reverted if TrkA inactivation during hypoxia is suppressed by the addition of NGF.Our results suggest that TrkA activation may play an important role in the survival of OT neurons subjected to acute hypoxia. The role of TrkA in neuronal survival after injury may be advantageously used for the generation of neuroprotective strategies to improve prenatal insult outcomes.

Involvement of sonic hedgehog and notch signaling in regenerative neurogenesis in adult zebrafish optic tectum after stab injury.

Unlike humans and other mammals, adult zebrafish have the superior capability to recover from central nervous system (CNS) injury. We previously found that proliferation of radial glia (RG) is induced in response to stab injury in optic tectum and that new neurons are generated from RG after stab injury. However, molecular mechanisms which regulate proliferation and differentiation of RG are not well known. In the present study, we investigated Shh and Notch signaling as potential mechanisms regulating regeneration in the optic tectum of adult zebrafish. We used Shh reporter fish and confirmed that canonical Shh signaling is activated specifically in RG after stab injury. Moreover, we have shown that Shh signaling promotes RG proliferation and suppresses their differentiation into neurons after stab injury. In contrast, Notch signaling was down-regulated after stab injury, indicated by the decrease in the expression level of her4 and her6, a target gene of Notch signaling. We also found that inhibition of Notch signaling after stab injury induced more proliferative RG, but that inhibition of Notch signaling inhibited generation of newborn neurons from RG after stab injury. These results suggest that high level of Notch signaling keeps RG quiescent and that appropriate level of Notch signaling is required for generation of newborn neurons from RG. Under physiological condition, activation of Shh signaling or inhibition of Notch signaling also induced RG proliferation. In adult optic tectum of zebrafish, canonical Shh signaling and Notch signaling play important roles in proliferation and differentiation of RG in physiological and regenerative conditions.

Environmentally induced changes to brain morphology predict cognitive performance.

The relationship between the size and structure of a species' brain and its cognitive capacity has long interested scientists. Generally, this work relates interspecific variation in brain anatomy with performance on a variety of cognitive tasks. However, brains are known to show considerable short-term plasticity in response to a range of social, ecological and environmental factors. Despite this, we have a remarkably poor understanding of how this impacts on an animal's cognitive performance. Here, we non-invasively manipulated the relative size of brain regions associated with processing visual and chemical information in fish (the optic tectum and olfactory bulbs, respectively). We then tested performance in a cognitive task in which information from the two sensory modalities was in conflict. Although the fish could effectively use both visual and chemical information if presented in isolation, when they received cues from both modalities simultaneously, those with a relatively better developed optic tectum showed a greater reliance on visual information, while individuals with relatively better developed olfactory bulbs showed a greater reliance on chemical information. These results suggest that short-term changes in brain structure, possibly resulting from an attempt to minimize the costs of developing unnecessary but energetically expensive brain regions, may have marked effects on cognitive performance.This article is part of the theme issue 'Causes and consequences of individual differences in cognitive abilities'.

Tectal CRFR1 receptors modulate food intake and feeding behavior in the South African clawed frog Xenopus laevis

The optic tectum and superior colliculus rapidly inhibit food intake when a visual threat is present. Previous work indicates that CRF, acting on CRFR1 receptors, may play a role in tectal inhibition of feeding behavior and food intake. Here we test the hypothesis that tectal CRFR1 receptors modulate food intake and feeding behavior in juvenile Xenopus laevis. We performed five experiments to test the following questions: 1) Does tectal CRF injection decrease food intake/feeding behavior? 2) Does a selective CRFR1 antagonist block CRF effects on feeding/feeding behavior? 3) Does a reactive stressor decrease food intake/feeding behavior? 4) Does a selective CRFR1 antagonist block reactive stress-induced decrease in feeding/feeding behavior? 5) Does food deprivation increase food intake/feeding behavior? Tectal CRF injections reduced food intake and influenced exploratory behavior, hindlimb kicks, and time in contact with food. These effects were blocked by the selective R1 antagonist NBI-27914. Exposure to a reactive stressor decreased food intake and this effect was blocked by NBI-27914. Neither food intake or feeding behavior changed following 1 wk of food deprivation. Overall, we conclude that activation of tectal CRFR1 inhibits food intake in juvenile X. laevis. Furthermore, tectal CRFR1 receptors appear to be involved in the reduction of food intake that occurs in response to a reactive stressor.

"Shepherd's crook" neurons drive and synchronize the enhancing and suppressive mechanisms of the midbrain stimulus selection network.

The optic tectum (TeO), or superior colliculus, is a multisensory midbrain center that organizes spatially orienting responses to relevant stimuli. To define the stimulus with the highest priority at each moment, a network of reciprocal connections between the TeO and the isthmi promotes competition between concurrent tectal inputs. In the avian midbrain, the neurons mediating enhancement and suppression of tectal inputs are located in separate isthmic nuclei, facilitating the analysis of the neural processes that mediate competition. A specific subset of radial neurons in the intermediate tectal layers relay retinal inputs to the isthmi, but at present it is unclear whether separate neurons innervate individual nuclei or a single neural type sends a common input to several of them. In this study, we used in vitro neural tracing and cell-filling experiments in chickens to show that single neurons innervate, via axon collaterals, the three nuclei that comprise the isthmotectal network. This demonstrates that the input signals representing the strength of the incoming stimuli are simultaneously relayed to the mechanisms promoting both enhancement and suppression of the input signals. By performing in vivo recordings in anesthetized chicks, we also show that this common input generates synchrony between both antagonistic mechanisms, demonstrating that activity enhancement and suppression are closely coordinated. From a computational point of view, these results suggest that these tectal neurons constitute integrative nodes that combine inputs from different sources to drive in parallel several concurrent neural processes, each performing complementary functions within the network through different firing patterns and connectivity.

Lateralized expression of left-right axis formation genes is shared by adult brains of lefty and righty scale-eating cichlids

Variation in the laterality often exists within species and can be maintained by frequency-dependent selection. Although the molecular developmental mechanisms underlying the left-right axis formation have been investigated, the genomic mechanisms underlying variation in laterality remain largely unknown. The scale-eating cichlid Perissodus microlepis in Lake Tanganyika exhibit lateralized predation; lefty individuals with the mouth opening toward the right preferentially attack on the prey's left trunk, while righty individuals with the opposite opening attacks on the right trunk. Here, we performed RNA-sequencing and subsequent confirmation with quantitative-PCR in the telencephalon, optic tectum, and hindbrain of the cichlid and identified five genes (pkd1b, ntn1b, ansn, pde6g, and rbp4l1) that were differentially expressed between the hemispheres regardless of the laterality. Surprisingly, pkd1b and ntn1b are involved in nodal and netrin signalling, respectively, which are important for left–right asymmetry formation during early embryogenesis. This result indicates that nodal- and netrin-related signals may also play important roles in the maintenance of asymmetry in adult brain. By contrast, no genes showed reversal of lateral differences between lefty and righty individuals in any brain regions examined, suggesting that laterality in the scale-eating cichlid does not simply result from inversion of the left–right asymmetry of gene expression.

Behavioral Evidence and Neural Correlates of Perceptual Grouping by Motion in the Barn Owl.

Perceiving an object as salient from its surround often requires a preceding process of grouping the object and background elements as perceptual wholes. In humans, motion homogeneity provides a strong cue for grouping, yet it is unknown to what extent this occurs in nonprimate species. To explore this question, we studied the effects of visual motion homogeneity in barn owls of both genders, at the behavioral as well as the neural level. Our data show that the coherency of the background motion modulates the perceived saliency of the target object. An object moving in an odd direction relative to other objects attracted more attention when the other objects moved homogeneously compared with when moved in a variety of directions. A possible neural correlate of this effect may arise in the population activity of the intermediate/deep layers of the optic tectum. In these layers, the neural responses to a moving element in the receptive field were suppressed when additional elements moved in the surround. However, when the surrounding elements all moved in one direction (homogeneously moving), they induced less suppression of the response compared with nonhomogeneously moving elements. Moreover, neural responses were more sensitive to the homogeneity of the background motion than to motion-direction contrasts between the receptive field and the surround. The findings suggest similar principles of saliency-by-motion in an avian species as in humans and show a locus in the optic tectum where the underlying neural circuitry may exist.SIGNIFICANCE STATEMENT A critical task of the visual system is to arrange incoming visual information to a meaningful scene of objects and background. In humans, elements that move homogeneously are grouped perceptually to form a categorical whole object. We discovered a similar principle in the barn owl's visual system, whereby the homogeneity of the motion of elements in the scene allows perceptually distinguishing an object from its surround. The novel findings of these visual effects in an avian species, which lacks neocortical structure, suggest that our basic visual perception shares more universal principles across species than presently thought, and shed light on possible brain mechanisms for perceptual grouping.

Nonapeptide levels in male cleaner fish brains during interactions with unfamiliar intra and interspecific partners

In nature, cleaner fish face complex situations arising from a highly demanding socio-environment that includes conspecific and mutualistic relationships. The nonapeptide arginine vasotocin (AVT) is known to play a relevant role in the expression of mutualistic behaviour in the Indo-Pacific bluestreak cleaner wrasse (Labroides dimidiatus). However, the role of the second nonapeptide isotocin (IT) in mediating social behaviour has yet to be established. Here we investigated whether there is a link between the distribution of active nonapeptides across brain regions and cleaners’ social behaviour. Male cleaner fish were exposed to different social contexts: (i) an unfamiliar male conspecific (conspecific context), (ii) an unfamiliar interspecific partner (blond naso tang, Naso elegans; mutualistic context) and (iii) a ball (non-social context). Furthermore, to distinguish between physicochemical and visual stimulations, cleaners were also exposed to an unfamiliar conspecific or an unfamiliar interspecific partner placed in a separate, smaller aquaria while inside the experimental tank. We then measured biologically available AVT and IT levels in distinct brain macro-areas, forebrain, optic tectum, cerebellum and brain stem, using high-performance liquid chromatography with fluorescence detection. We did not find any association between the levels of AVT and social contexts. On the other hand, we found lower levels of IT in the forebrain of cleaners in contact with a conspecific compared to those introduced to a client and a client inside another aquarium. Additionally, cleaners displayed aggression but only towards other conspecifics. These findings indicate that (i) IT in cleaners’ forebrain is linked with mutualistic engagement between cleaners and clients, (ii) stimulation of IT pathways in the forebrain is probably linked with the visual recognition of potential clients and (iii) physicochemical signals are essential to trigger aggressive displays in male conspecifics. The visual stimulation of IT pathways in the forebrain of male cleaner wrasses while introduced to sympatric clients is associated with the establishment of cleaner-client mutualistic relationship; physicochemical signals are important to trigger aggressive displays in male cleaners introduced to conspecifics.

Luminance Changes Drive Directional Startle through a Thalamic Pathway

Looming visual stimuli result in escape responses that are conserved from insects to humans. Despite their importance for survival, the circuits mediating visual startle have only recently been explored in vertebrates. Here we show that the zebrafish thalamus is a luminance detector critical to visual escape. Thalamic projection neurons deliver dim-specific information to the optic tectum, and ablations of these projections disrupt normal tectal responses to looms. Without this information, larvae are less likely to escape from dark looming stimuli and lose the ability to escape away from the source of the loom. Remarkably, when paired with an isoluminant loom stimulus to the opposite eye, dimming is sufficient to increase startle probability and to reverse the direction of the escape so that it is toward the loom. We suggest that bilateral comparisons of luminance, relayed from the thalamus to the tectum, facilitate escape responses and are essential for their directionality.

Principles of Functional Circuit Connectivity: Insights From Spontaneous Activity in the Zebrafish Optic Tectum

The brain is continuously active, even in the absence of external stimulation. In the optic tectum of the zebrafish larva, this spontaneous activity is spatially organized and reflects the circuit’s functional connectivity. The structure of the spontaneous activity displayed patterns associated with aspects of the larva’s preferences when engaging in complex visuo-motor behaviors, suggesting that the tectal circuit is adapted for the circuit’s functional role in detecting visual cues and generating adequate motor behaviors. Further studies in sensory deprived larvae suggest that the basic structure of the functional connectivity patterns emerges even in the absence of retinal inputs, but that its fine structure is affected by visual experience.

Eyes Wide Shut: the impact of dim-light vision on neural investment in marine teleosts.

Understanding how organismal design evolves in response to environmental challenges is a central goal of evolutionary biology. In particular, assessing the extent to which environmental requirements drive general design features among distantly related groups is a major research question. The visual system is a critical sensory apparatus that evolves in response to changing light regimes. In vertebrates, the optic tectum is the primary visual processing centre of the brain and yet it is unclear how or whether this structure evolves while lineages adapt to changes in photic environment. On one hand, dim-light adaptation is associated with larger eyes and enhanced light-gathering power that could require larger information processing capacity. On the other hand, dim-light vision may evolve to maximize light sensitivity at the cost of acuity and colour sensitivity, which could require less processing power. Here, we use X-ray microtomography and phylogenetic comparative methods to examine the relationships between diel activity pattern, optic morphology, trophic guild and investment in the optic tectum across the largest radiation of vertebrates-teleost fishes. We find that despite driving the evolution of larger eyes, enhancement of the capacity for dim-light vision generally is accompanied by a decrease in investment in the optic tectum. These findings underscore the importance of considering diel activity patterns in comparative studies and demonstrate how vision plays a role in brain evolution, illuminating common design principles of the vertebrate visual system.

Ontogeny of Gonadotropin-Inhibitory Hormone (GnIH) in the cichlid fish Cichlasoma dimerus.

RFamide peptides are expressed in the early stages of development in most vertebrates. Gonadotropin-inhibitory hormone (GnIH) belongs to the RFamide family, and its role in reproduction has been widely studied in adult vertebrates, ranging from fish to mammals. As only three reports evaluated GnIH during development, the aim of this study was to characterise the ontogeny of GnIH in a fish model, Cichlasoma dimerus. We detected the presence of two GnIH-immunoreactive (GnIH-ir) cell clusters with spatial and temporal differences. One cluster was observed by 3 days post-hatching (dph) in the nucleus olfacto-retinalis (NOR) and the other in the nucleus posterioris periventricularis by 14 dph. The number of GnIH-ir neurons increased in both nuclei, whereas their size increased only in the NOR from hatchling to juvenile stages. These changes occurred from the moment larvae started feeding exogenously and during development and differentiation of gonadal primordia. We showed by double-label immunofluorescence that only GnIH-ir neurons in the NOR co-expressed GnRH3 associated peptide. In addition, GnIH-ir fibre density increased in all brain regions from 5 dph. GnIH-ir fibres were also detected in the retina, optic tract and optic tectum, suggesting that GnIH acts as a neuromodulator of photoreception and the integration of different sensory modalities. Also, there were GnIH-ir fibres in the pituitary from 14 dph, which were in close association with somatotropes. Moreover, GnIH-ir fibres were observed in the saccus vasculosus from 30 dph, suggesting a potential role of GnIH in the modulation of its function. Finally, we found that gnih was expressed from 1 dph, and that the pattern of variation of its transcript levels was in accordance with that of cell number. Present results are the starting point for the study of new GnIH roles during development. This article is protected by copyright. All rights reserved.

Nox2/cybb Deficiency Affects Zebrafish Retinotectal Connectivity.

NADPH oxidase (Nox)-derived reactive oxygen species (ROS) have been linked to neuronal polarity, axonal outgrowth, cerebellar development, regeneration of sensory axons, and neuroplasticity. However, the specific roles that individual Nox isoforms play during nervous system development in vivo remain unclear. To address this problem, we investigated the role of Nox activity in the development of retinotectal connections in zebrafish embryos. Zebrafish broadly express four nox genes (nox1, nox2/cybb, nox5, and duox) throughout the CNS during early development. Application of a pan-Nox inhibitor, celastrol, during the time of optic nerve (ON) outgrowth resulted in significant expansion of the ganglion cell layer (GCL), thinning of the ON, and a decrease in retinal axons reaching the optic tectum (OT). With the exception of GCL expansion, these effects were partially ameliorated by the addition of H2O2, a key ROS involved in Nox signaling. To address isoform-specific Nox functions, we used CRISPR/Cas9 to generate mutations in each zebrafish nox gene. We found that nox2/cybb chimeric mutants displayed ON thinning and decreased OT innervation. Furthermore, nox2/cybb homozygous mutants (nox2/cybb-/-) showed significant GCL expansion and mistargeted retinal axons in the OT. Neurite outgrowth from cultured zebrafish retinal ganglion cells was reduced by Nox inhibitors, suggesting a cell-autonomous role for Nox in these neurons. Collectively, our results show that Nox2/Cybb is important for retinotectal development in zebrafish.SIGNIFICANCE STATEMENT Most isoforms of NADPH oxidase (Nox) only produce reactive oxygen species (ROS) when activated by an upstream signal, making them ideal candidates for ROS signaling. Nox enzymes are present in neurons and their activity has been shown to be important for neuronal development and function largely by in vitro studies. However, whether Nox is involved in the development of axons and formation of neuronal connections in vivo has remained unclear. Using mutant zebrafish embryos, this study shows that a specific Nox isoform, Nox2/Cybb, is important for the establishment of axonal connections between retinal ganglion cells and the optic tectum.

Distribution of vesicular glutamate transporters in the brain of the turtle (Pseudemys scripta elegans).

The distribution of glutamatergic neurons has been extensively studied in mammalian and avian brains, but its distribution in a reptilian brain remains unknown. In the present study, the distribution of subpopulations of glutamatergic neurons in the turtle brain was examined by in situ hybridization using probes for vesicular glutamate transporter (VGLUT) 1-3. Strong VGLUT1 expression was observed in the telencephalic pallium; the mitral cells of the olfactory bulb, the medial, dorsomedial, dorsal, and lateral parts of the cerebral cortex, pallial thickening, and dorsal ventricular ridge; and also, in granule cells of the cerebellar cortex. Moderate to weak expression was found in the lateral and medial amygdaloid nuclei, the periventricular cellular layer of the optic tectum, and in some brainstem nuclei. VGLUT2 was weakly expressed in the telencephalon but was intensely expressed in the dorsal thalamic nuclei, magnocellular part of the isthmic nucleus, brainstem nuclei, and the rostral cervical segment of the spinal cord. The cerebellar cortex was devoid of VGLUT2 expression. The central amygdaloid nucleus did not express VGLUT1 or VGLUT2. VGLUT3 was localized in the parvocellular part of the isthmic nucleus, superior and inferior raphe nuclei, and cochlear nucleus. Our results indicate that the distribution of VGLUTs in the turtle brain is similar to that in the mammalian brain rather than that in the avian brain.

An ascending visual pathway to the dorsal telencephalon through the optic tectum and nucleus prethalamicus in the yellowfin goby Acanthogobius flavimanus (Temminck & Schlegel, 1845).

Dual visual pathways reaching the telencephalon appear to be an ancient vertebrate trait, but some teleost fish seem to possess only one pathway via the optic tectum. We undertook the present study to determine if and when this loss occurred during evolution. Tracer injection experiments to the optic nerve, the optic tectum, and the dorsal telencephalon were performed in the present study, to investigate ascending visual pathways to the dorsal telencephalon in an acanthopterygian teleost, the yellowfin goby Acanthogobius flavimanus (Temminck & Schlegel, 1845). We confirmed the presence of a nucleus prethalamicus (PTh) in the goby, which has been convincingly identified only in holocentrids, suggesting that this nucleus is present in other acanthopterygians. We found that the optic tectum projects to the PTh bilaterally. The PTh projects in turn to the dorsal telencephalon, ipsilaterally. These results suggest that the yellowfin goby possesses only an extrageniculate-like pathway, while a geniculate-like pathway could not be identified. This situation is common with that of holocentrids and may be a character common in acanthopterygians. It is possible that a geniculate-like system was lost in the common ancestor of acanthopterygians, although the scenario for the evolution of ascending visual systems in actinopterygians remains uncertain due to the lack of precise knowledge in a number of actinopterygian taxons.

Effects of early eye removal on the morphology of a multisensory neuron in the chicken optic tectum

The midbrain is a subcortical area involved in central functions such as integrating sensory modalities, movement initiation and bottom-up and top-down attention. In chicken, the midbrain roof is termed optic tectum (TeO) and consists of 15 layers with distinct in- and output regions. Visual input targets the superficial layers, while auditory input terminates in deeper layers. It has been shown that ablation of sensory epithelia leads to changes in the cellular patterning and structural organization of the sensory pathways. For the tectum, ablation of the eye anlagen was shown to affect retinorecipient neurons. While the gross morphology remained intact after enucleation, the shape of dendritic endings was changed presumably due to missing presynaptic input during synaptic pruning.We investigated the effect of deafferentation in a multisensory cell type, the Shepherd’s crook neuron (SCN) in the TeO. SCNs have distinct dendritic branches in retinorecipient layers (superficial layers 1 to 5 and 7) and in layers where auditory input terminates. To assess whether removal of a single sensory input only affects the dendrites recipient for that input, we removed the eye anlagen and retrogradely labeled SCNs later in embryogenesis to visualize the morphology in lesioned and non-lesioned embryos. We found no changes in the gross morphology or in the basal dendrites, but an altered growth of the fine structures at the apical dendrite of SCNs in the retinorecipient layers. Our data indicate that the neuronal morphology of SCNs is mostly predefined before retinal innervation affect the fine structure.

Optical interrogation of neuronal circuitry in zebrafish using genetically encoded voltage indicators

Optical measurement of membrane potentials enables fast, direct and simultaneous detection of membrane potentials from a population of neurons, providing a desirable approach for functional analysis of neuronal circuits. Here, we applied recently developed genetically encoded voltage indicators, ASAP1 (Accelerated Sensor of Action Potentials 1) and QuasAr2 (Quality superior to Arch 2), to zebrafish, an ideal model system for studying neurogenesis. To achieve this, we established transgenic lines which express the voltage sensors, and showed that ASAP1 is expressed in zebrafish neurons. To examine whether neuronal activity could be detected by ASAP1, we performed whole-cerebellum imaging, showing that depolarization was detected widely in the cerebellum and optic tectum upon electrical stimulation. Spontaneous activity in the spinal cord was also detected by ASAP1 imaging at single-cell resolution as well as at the neuronal population level. These responses mostly disappeared following treatment with tetrodotoxin, indicating that ASAP1 enabled optical measurement of neuronal activity in the zebrafish brain. Combining this method with other approaches, such as optogenetics and behavioural analysis may facilitate a deeper understanding of the functional organization of brain circuitry and its development.