Masae Kinoshita

The Habenulo-Raphe Serotonergic Circuit Encodes an Aversive Expectation Value Essential for Adaptive Active Avoidance of Danger

Anticipation of danger at first elicits panic in animals, but later it helps them to avoid the real threat adaptively. In zebrafish, as fish experience more and more danger, neurons in the ventral habenula (vHb) showed tonic increase in the activity to the presented cue and activated serotonergic neurons in the median raphe (MR). This neuronal activity could represent the expectation of a dangerous outcome and be used for comparison with a real outcome when the fish is learning how to escape from a dangerous to a safer environment. Indeed, inhibiting synaptic transmission from vHb to MR impaired adaptive avoidance learning, while panic behavior induced by classical fear conditioning remained intact. Furthermore, artificially triggering this negative outcome expectation signal by optogenetic stimulation of vHb neurons evoked place avoidance behavior. Thus, vHb-MR circuit is essential for representing the level of expected danger and behavioral programming to adaptively avoid potential hazard.

Social conflict resolution regulated by two dorsal habenular subregions in zebrafish

How to win a fish fight When to cease aggression and escape is an important decision that fighting animals must make. Chou et al. characterized the role of two nuclei in a brain area of the zebrafish called the dorsal habenula (dHb) during social aggression (see the Perspective by Desban and Wyart). Silencing the lateral dHb reduced the likelihood of winning a fight, whereas silencing the medial dHb increased the likelihood of winning. Thus, these two nuclei antagonistically control the threshold for surrender. Science, this issue p. 87; see also p. 42 The neuronal basis for keeping the aggression of fighting fish in check is elucidated. [Also see Perspective by Desban and Wyart] When animals encounter conflict they initiate and escalate aggression to establish and maintain a social hierarchy. The neural mechanisms by which animals resolve fighting behaviors to determine such social hierarchies remain unknown. We identified two subregions of the dorsal habenula (dHb) in zebrafish that antagonistically regulate the outcome of conflict. The losing experience reduced neural transmission in the lateral subregion of dHb (dHbL)–dorsal/intermediate interpeduncular nucleus (d/iIPN) circuit. Silencing of the dHbL or medial subregion of dHb (dHbM) caused a stronger predisposition to lose or win a fight, respectively. These results demonstrate that the dHbL and dHbM comprise a dual control system for conflict resolution of social aggression.

Imaging of Neural Ensemble for the Retrieval of a Learned Behavioral Program

The encoding of long-term associative memories for learned behaviors is a fundamental brain function. Yet, how behavior is stably consolidated and retrieved in the vertebrate cortex is poorly understood. We trained zebrafish in aversive reinforcement learning and measured calcium signals across their entire brain during retrieval of the learned response. A discrete area of dorsal telencephalon that was inactive immediately after training became active the next day. Analysis of the identified area indicated that it was specific and essential for long-term memory retrieval and contained electrophysiological responses entrained to the learning stimulus. When the behavioral rule changed, a rapid spatial shift in the functional map across the telencephalon was observed. These results demonstrate that the retrieval of long-term memories for learned behaviors can be studied at the whole-brain scale in behaving zebrafish in vivo. Moreover, the findings indicate that consolidated memory traces can be rapidly modified during reinforcement learning.

Spontaneous depolarization waves of multiple origins in the embryonic rat CNS.

During development, correlated neuronal activity plays an important role in the establishment of the central nervous system (CNS). We have previously reported that a widely propagating correlated neuronal activity, termed the depolarization wave, is evoked by various sensory inputs. A remarkable feature of the depolarization wave is that it spreads broadly through the brain and spinal cord. In the present study, we examined whether the depolarization wave occurs spontaneously in the embryonic rat CNS and, if so, where it originates. In E15–16 rat embryos, spontaneous optically‐revealed signals appeared in association with the rhythmic discharges of cranial motoneurons and propagated widely with similar characteristics to the evoked depolarization wave. At E15, the spontaneous wave mostly originated in the cervical to upper lumbar cords. At E16, the wave was predominantly generated in the lumbosacral cord although a wave associated with the second oscillatory burst was initiated in the rostral cord. At E16, a few waves also originated in the rostral ventrolateral medulla and the dorsomedial pons. When the influence of the caudal cord was removed by transecting the spinal cord, the contribution of the medulla and pons became more significant. These results show that the depolarization wave can be triggered by the spontaneous activity of multiple neuronal populations which are distributed widely from the pons to the lumbosacral cord, although the spinal cord usually plays a predominant role. This network possibly works as a self‐distributing system that maintains the incidence and complicated patterns of the correlated activity in the developing CNS.

Origin of the earliest correlated neuronal activity in the chick embryo revealed by optical imaging with voltage-sensitive dyes.

Spontaneous correlated neuronal activity during early development spreads like a wave by recruiting a large number of neurons, and is considered to play a fundamental role in neural development. One important and as yet unresolved question is where the activity originates, especially at the earliest stage of wave expression. In other words, which part of the brain differentiates first as a source of the correlated activity, and how does it change as development proceeds? We assessed this issue by examining the spatiotemporal patterns of the depolarization wave, the optically identified primordial correlated activity, using the optical imaging technique with voltage‐sensitive dyes. We surveyed the region responsible for the induction of the evoked and spontaneous depolarization waves in chick embryos, and traced its developmental changes. The results showed that the wave initially originated in a restricted area near the obex and was generated by multiple regions at later stages. We suggest that the upper cervical cord/lower medulla near the obex is the kernel that differentiates first as the source of the correlated activity, and that regional and temporal differences in neuronal excitability might underlie the developmental profile of wave generation in early chick embryos.

Neuromodulatory effects of gonadotropin-releasing hormone on retinotectal synaptic transmission in the optic tectum of rainbow trout

Gonadotropin‐releasing hormone (GnRH) is a hypophysiotropic decapeptide that stimulates the release of gonadotropins from the pituitary. In addition, there are extra‐hypothalamic GnRH neurons that project to all regions of the brain and whose function remains unknown. Here, we investigated the effects of GnRH on retinotectal synaptic transmission, the synapses of which are formed between retinal fibers and tectal periventricular neurons that express GnRH receptor mRNA. We used rainbow trout as our study model. The excitatory postsynaptic currents (EPSCs), which were evoked by electrical stimulation of the retinal fibers and recorded in periventricular neurons, were suppressed by antagonists of ionotropic glutamate receptors. EPSCs were increased by application of each of two types of GnRH (GnRH2 and GnRH3) in the trout tectum. Such facilitation lasted for at least 10 min after application of the GnRH. To our knowledge, this is the first report of GnRH modulating conventional synaptic transmission in the brain, suggesting that tectal GnRH enhances tectal sensitivity for retinal inputs. Furthermore, such long‐lasting facilitation might occur across all the brain regions innervated by GnRH neurons, and GnRH might simultaneously switch neuronal activities in the brain regions relevant to reproductive behaviors.

Retinotectal transmission in the optic tectum of rainbow trout

Retinotectal transmission has not yet been well characterized at the cellular level in the optic tectum. To address this issue, we used a teleost, the rainbow trout, and characterized periventricular neurons as postsynaptic cells expected to receive the retinotectal inputs to the optic tectum. The somata of periventricular neurons are localized in the upper zone of the stratum periventriculare (SPV), whereas the lower zone of the SPV comprises the cell body layer of radial glial cells. Ca2+ imaging identified functional ionotropic glutamate receptors in periventricular neurons. We also cloned cDNAs encoding the NR1 subunit of N‐methyl‐D‐aspartic acid (NMDA) receptors and the GluR2 subunit of (±)‐α‐amino‐3‐hydroxy‐5‐methyl‐isoxazole‐4‐propionic acid (AMPA) receptors, and detected their mRNAs in periventricular neurons by in situ hybridization. The presence of the receptor subunit proteins was also confirmed in the dendrites of periventricular neurons by immunoblotting and immunohistochemistry. On the other hand, radial glial cells in the lower zone of the SPV did not respond to glutamate applications, and mRNA and immunoreactivities of ionotropic glutamate receptors were not detected in glial cells. The present findings suggest that glutamatergic transmission at synapses between retinotectal afferents and periventricular neurons is mediated by the functional NMDA and AMPA receptors. J. Comp. Neurol. 484:249–259, 2005.

Periventricular efferent neurons in the optic tectum of rainbow trout

The efferent connections and axonal and dendritic morphologies of periventricular neurons were examined in the optic tectum of rainbow trout to classify periventricular efferent neurons in salmonids. Among the target nuclei of tectal efferents, tracer injections to the following four structures labeled periventricular neurons: the area pretectalis pars dorsalis (APd), nucleus pretectalis superficialis pars magnocellularis (PSm), nucleus ventrolateralis of torus semicircularis (TS), and nucleus isthmi (NI). Two types of periventricular neurons were labeled by injections to the APd. One of them had an apical dendrite ramifying at the stratum fibrosum et griseum superficiale (SFGS), with an axon that bifurcated into two branches at the stratum griseum centrale (SGC), and the other had an apical dendrite ramifying at the SGC. Two types of periventricular neurons were labeled after injections to the TS. One of them had an apical dendrite ramifying at the boundary between the stratum opticum (SO) and the SFGS, and the other had dendritic branches restricted to the stratum album centrale or stratum periventriculare. Injections to the PSm and NI labeled periventricular neurons of the same type with an apical dendrite ramifying at the SO and a characteristic axon that split into superficial and deep branches projecting to the PSm and NI, respectively. This cell type also possessed axonal branches that terminated within the tectum. These results indicate that periventricular efferent neurons can be classified into at least five types that possess type‐specific axonal and dendritic morphologies. We also describe other tectal neurons labeled by the present injections. J. Comp. Neurol. 499:546–564, 2006.

Roles of periventricular neurons in retinotectal transmission in the optic tectum

The midbrain roof is a retinorecipient region referred to as the optic tectum in lower vertebrates, and the superior colliculus in mammals. The retinal fibers projecting to the tectum transmit visual information to tectal retinorecipient neurons. Periventricular neurons are a subtype of these neurons that have their somata in the deepest layer of the teleostean tectum and apical dendrites ramifying at more superficial layers consisting of retinal fibers. The retinotectal synapses between the retinal fibers and periventricular neurons are glutamatergic, and ionotropic glutamate receptors mediate the transmission in these synapses. This transmission involves long-term potentiation, and is modulated by hormone action. Visual information processed in the periventricular neurons is transmitted to adjacent tectal cells and target nuclei of periventricular neuron axonal branches, some of which relay the visual information to other brain areas controlling behavior. We demonstrated that periventricular neurons play a principal role in visual information processing in the teleostean optic tectum; the effects of tectal output on behavior is discussed also in the present review.

Multiple-site optical recording for characterization of functional synaptic organization of the optic tectum of rainbow trout

To map the functional synaptic organization over a wide area in the optic tectum, we directly monitored two‐dimensional propagation of postsynaptic depolarization evoked by firing of retinotectal afferents in optic tectum slices prepared from rainbow trout (Oncorhynchus mykiss), using a voltage‐sensitive dye and a photodiode array system. The postsynaptic responses to afferent stimulation first propagated in the stratum opticum and stratum fibrosum et griseum superficiale in an anterograde fashion in the afferents and then expanded vertically into the deep layers. This vertical propagation appeared to occur along a bundle‐like structure that corresponded well with a cluster of neurons whose somata are located in the stratum periventriculare. Pharmacological studies showed that these postsynaptic responses were mediated by ionotropic glutamate receptors. On the other hand, the optical signals appeared to consist of at least two components (a transient signal and a slow signal). The second transient signal summated with the first slow signal by paired stimulation, suggesting that the transient and slow signals originated from different cell types. Taken together, these results showed that the functional synaptic organization of the teleost optic tectum comprises of two depolarization‐signal propagating paths along a horizontal layer structure and a vertical bundle‐like structure and that these synaptic responses occur via glutamatergic transmission.