During the embryonic period, neuronal communication starts before the establishment of the synapses with alternative forms of neuronal excitability, called here embryonic neural excitability (ENE). ENE has been shown to modulate the unfolding of development transcriptional programs, but the global consequences for developing organisms are not all understood. Here, we monitored calcium (Ca2+) transients in the telencephalon of zebrafish embryos as a proxy for ENE to assess the efficacy of transient pharmacological treatments to either increase or decrease ENE. Increasing or decreasing ENE at the end of the embryonic period promoted an increase or a decrease in the numbers of dopamine (DA) neurons, respectively. This plasticity of dopaminergic specification occurs in the subpallium (SP) of zebrafish larvae at 6 d postfertilization (dpf), within a relatively stable population of vMAT2-positive cells. Nondopaminergic vMAT2-positive cells hence constitute an unanticipated biological marker for a reserve pool of DA neurons that can be recruited by ENE. Modulating ENE also affected larval locomotion several days after the end of the treatments. In particular, the increase of ENE from 2 to 3 dpf promoted hyperlocomotion of larvae at 6 dpf, reminiscent of zebrafish endophenotypes reported for attention deficit hyperactivity disorders (ADHDs). These results provide a convenient framework for identifying environmental factors that could disturb ENE as well as to study the molecular mechanisms linking ENE to neurotransmitter specification.

The scientific context of the special issue dedicated to Luis Puelles and Agustin Gonzalez is given in this short epilogue.

Summary This review of the literature on varietal change in sub‐Saharan Africa looks in detail at adoption of new varieties of bananas in Uganda, cassava in Nigeria, potato in Kenya, sweetpotato in Uganda and yams in Côte d’Ivoire. The review explored three hypotheses about drivers of varietal change. There was a strong confirmation for the hypothesis that insufficient priority given to consumer‐preferred traits by breeding programmes contributes to the limited uptake of modern varieties (MVs) and low varietal turnover. Lack of evidence meant the second hypothesis of insufficient attention to understanding and responding to gender differences in consumer preferences for quality and post‐harvest traits was unresolved. The evidence on the third hypothesis about the informal seed system contributing to slow uptake of MVs was mixed. In some cases, the informal system has contributed to rapid uptake of MVs, but often it appears to be a barrier with inconsistent varietal naming a major challenge.

The frontal cortex of mammals has been the site of significant reshuffles during the about 310 million years of mammalian evolution. The existence of the frontal lobe is linked to the emergence of major cognitive and motor abilities, from the elaboration of efficient problem-solving strategies to the use of language in human. However, presenting the frontal lobe of the human cortex in an evolutionary perspective is a real challenge, almost a wager lost in advance. Reasons are the following. Although present in all the mammalian species known so far, the frontal lobe, and especially its prefrontal domain, is one of the most variable brain regions among mammalian species, even among individuals. The prefrontal cortex (PFC) exhibits a lot of variations not only in size (from 35% of the total brain volume in human to less than 4% in the cat for example [1]), but also in its connections, cell composition, branching pattern of neuronal processes, etc. Similar remarks can be made for other domains of the frontal cortex, including the motor and Broca's area for example. These variations do not fit with the phylogenetical position of the corresponding species, even in hominins, precluding any elaboration of a simple scenario of the evolution of the frontal cortex in mammals. Although disappointing at first glance, this observation points to the major influence of environment and lifestyle in shaping the functional anatomy of the frontal lobe during evolution. This has been rendered possible both by the extraordinary plasticity of the development of the frontal lobe, especially of the PFC, and by strong functional selection upon demanding environmental constraints. This is well exemplified by the discrepancies between overall brain volume and the size of the PFC in species like primates, cetaceans or carnivores [2]. Despite these large variations, some building principles seem to be at work to support the main functions of the frontal cortex. The frontal lobe includes the main motor area of the brain in all mammals, with different specializations. In contrast, the PFC is mostly an associative area receiving multimodal sensory cues and establishing connections with the basal ganglia, amygdala, insular and medial temporal cortex, all features conserved in mammals. Its orbito-frontal part is often defined by its afferent connections from the mediodorsal nucleus of thalamus. Then, it receives strong dopaminergic input from the mesocortical pathway, playing a crucial role in executive functions and motor planning. By definition, a PFC does not exist in species other than mammals, since only mammals have a six-layered cortical pallium. It is thus very striking that non-mammalian amniotes such as birds also display high cognitive flexibility and skills qualified as “intelligence”, and also often use languages in social communication, all functions that require a frontal cortex in mammals. In birds, and probably in other vertebrate species also, the same rules of functional organization of the pallium as those found for the mammalian cortex seem to operate and underlie similar sophisticated behaviors. In particular, the identification in some birds of a pallial structure exhibiting most of the main characteristics of the mammalian PFC is striking. Since this pallial structure is not homologous to the PFC in any respect, it strongly suggests that these executive structures have emerged independently during evolution [3]. Thus, very likely, similar functional constraints led the pallium of mammals and birds to converge on similar – yet independent – rules to build “executive” brain areas. Similar convergence holds true for the neural circuits underlying speech and languages, also in birds and primates. These types of convergences certainly exist in other groups of vertebrates such as teleost fishes. The frontal cortex is thus unique to mammals, but not the only one of its kind to have emerged during the evolution of the different vertebrate groups.

Cerebrospinal fluid‐contacting (CSF‐c) cells containing monoamines such as dopamine (DA) and serotonin (5‐HT) occur in the periventricular zones of the hypothalamic region of most vertebrates except for placental mammals. Here we compare the organization of the CSF‐c cells in chicken, Xenopus, and zebrafish, by analyzing the expression of synthetic enzymes of DA and 5‐HT, respectively, tyrosine hydroxylase (TH) and tryptophan hydroxylase (TPH), and draw an evolutionary scenario for this cell population. Due to the lack of TH immunoreactivity in this region, the hypothalamic CSF‐c cells have been thought to take up DA from the ventricle instead of synthesizing it. We demonstrate that a second TH gene (TH2) is expressed in the CSF‐c cells of all the three species, suggesting that these cells do indeed synthetize DA. Furthermore, we found that many CSF‐c cells coexpress TH2 and TPH1 and contain both DA and 5‐HT, a dual neurotransmitter phenotype hitherto undescribed in the brain of any vertebrate. The similarities of CSF‐c cells in chicken, Xenopus, and zebrafish suggest that these characteristics are inherited from the common ancestor of the Osteichthyes. A significant difference between tetrapods and teleosts is that teleosts possess an additional CSF‐c cell population around the posterior recess (PR) that has emerged in specific groups of Actinopterygii. Our comparative analysis reveals that the hypothalamus in mammals and teleosts has evolved in a divergent manner: placental mammals have lost the monoaminergic CSF‐c cells, while teleosts have increased their relative number.

In the current model, the most anterior part of the forebrain (secondary prosencephalon) is subdivided into the telencephalon dorsally and the hypothalamus ventrally. Our recent study identified a new morphogenetic unit named the optic recess region (ORR) between the telencephalon and the hypothalamus. This modification of the forebrain regionalization based on the ventricular organization resolved some previously unexplained inconsistency about regional identification in different vertebrate groups. The ventricular-based comparison also revealed a large diversity within the subregions (notably in the hypothalamus and telencephalon) among different vertebrate groups. In tetrapods there is only one hypothalamic recess, while in teleosts there are two recesses. Most notably, the mammalian and teleost hypothalami are two extreme cases: the former has lost the cerebrospinal fluid-contacting (CSF-c) neurons, while the latter has increased them. Thus, one to one homology of hypothalamic subregions in mammals and teleosts requires careful verification. In the telencephalon, different developmental processes between Sarcopterygii (lobe-finned fish) and Actinopterygii (ray-finned fish) have already been described: the evagination and the eversion. Although pallial homology has been long discussed based on the assumption that the medial-lateral organization of the pallium in Actinopterygii is inverted from that in Sarcopterygii, recent developmental data contradict this assumption. Current models of the brain organization are largely based on a mammalian-centric point of view, but our comparative analyses shed new light on the brain organization of Osteichthyes.

The quantitative and systematic analysis of embryonic cell dynamics from in vivo 3D+time image data sets is a major challenge at the forefront of developmental biology. Despite recent breakthroughs in the microscopy imaging of living systems, producing an accurate cell lineage tree for any developing organism remains a difficult task. We present here the BioEmergences workflow integrating all reconstruction steps from image acquisition and processing to the interactive visualization of reconstructed data. Original mathematical methods and algorithms underlie image filtering, nucleus centre detection, nucleus and membrane segmentation, and cell tracking. They are demonstrated on zebrafish, ascidian and sea urchin embryos with stained nuclei and membranes. Subsequent validation and annotations are carried out usingMov-IT, a custom-made graphical interface. Compared with eight other software tools, our workflow achieved the best lineage score. Delivered in standalone or web service mode, BioEmergences and Mov-IT offer a unique set of tools for in silico experimental embryology.

Traditional yam-based cropping systems (shifting cultivation, slash-and-burn, and short fallow) often result in deforestation and soil nutrient depletion. The objective of this study was to determine the impact of yam-based systems with herbaceous legumes on dry matter (DM) production (tubers, shoots), nutrients removed and recycled, and the soil fertility changes. We compared smallholders’ traditional systems (1-year fallow of Andropogon gayanus-yam rotation, maize-yam rotation) with yam-based systems integrated herbaceous legumes (Aeschynomene histrix/maize intercropping-yam rotation, Mucuna pruriens/maize intercropping-yam rotation). The experiment was conducted during the 2002 and 2004 cropping seasons with 32 farmers, eight in each site. For each of them, a randomized complete block design with four treatments and four replicates was carried out using a partial nested model with five factors: Year, Replicate, Farmer, Site, and Treatment. Analysis of variance (ANOVA) using the general linear model (GLM) procedure was applied to the dry matter (DM) production (tubers, shoots), nutrient contribution to the systems, and soil properties at depths 0–10 and 10–20 cm. DM removed and recycled, total N, P, and K recycled or removed, and soil chemical properties (SOM, N, P, K, and pH water) were significantly improved on yam-based systems with legumes in comparison with traditional systems.

Dopamine neurotransmission regulates various brain functions, and its regulatory roles are mediated by two families of G protein-coupled receptors: the D1 and D2 receptor families. In mammals, the D1 family comprises two receptor subtypes (D1 and D5), while the D2 family comprises three receptor subtypes (D2, D3 and D4). Phylogenetic analyses of dopamine receptor genes strongly suggest that the common ancestor of Osteichthyes (bony jawed vertebrates) possessed four subtypes in the D1 family and five subtypes in the D2 family. Mammals have secondarily lost almost half of the ancestral dopamine receptor genes, whereas nonmammalian species kept many of them. Although the mammalian situation is an exception among Osteichthyes, the current classification and characterization of dopamine receptors are based on mammalian features, which have led to confusion in the identification of dopamine receptor subtypes in nonmammalian species. Here we begin by reviewing the history of the discovery of dopamine receptors in vertebrates. The recent genome sequencing of coelacanth, gar and elephant shark led to the proposal of a refined scenario of evolution of dopamine receptor genes. We also discuss a current problem of nomenclature of dopamine receptors. Following the official nomenclature of mammalian dopamine receptors from D1 to D5, we propose to name newly identified receptor subtypes from D6 to D9 in order to facilitate the use of an identical name for orthologous genes among different species. To promote a nomenclature change which allows distinguishing the two dopamine receptor families, a nomenclature consortium is needed. This comparative perspective is crucial to correctly interpret data obtained in animal studies on dopamine-related brain disorders, and more fundamentally, to understand the characteristics of dopamine neurotransmission in vertebrates.

Regionalization is a critical, highly conserved step in the development of the vertebrate brain. Discrepancies exist in how regionalization of the anterior vertebrate forebrain is conceived since the `` preoptic area'' is proposed to be a part of the telencephalon in tetrapods but not in teleost fish. To gain insight into this complex morphogenesis, formation of the anterior forebrain was analyzed in 3D over time in zebrafish embryos, combining visualization of proliferation and differentiation markers, with that of developmental genes. We found that the region containing the preoptic area behaves as a coherent morphogenetic entity, organized around the optic recess and located between telencephalon and hypothalamus. This optic recess region (ORR) makes clear borders with its neighbor areas and expresses a specific set of genes (dlx2a, sim1a and otpb). We thus propose that the anterior forebrain (secondary prosencephalon) in teleosts contains three morphogenetic entities (telencephalon, ORR and hypothalamus), instead of two (telencephalon and hypothalamus). The ORR in teleosts could correspond to "telencephalic stalk area'' and `` alar hypothalamus'' in tetrapods, resolving current inconsistencies in the comparison of basal forebrain among vertebrates.

Dopaminergic (DA) neurons located in the preoptico-hypothalamic region of the brain exert a major neuroendocrine control on reproduction, growth, and homeostasis by regulating the secretion of anterior pituitary (or adenohypophysis) hormones. Here, using a retrograde tract tracing experiment, we identified the neurons playing this role in the zebrafish. The DA cells projecting directly to the anterior pituitary are localized in the most anteroventral part of the preoptic area, and we named them preoptico-hypophyseal DA (POHDA) neurons. During development, these neurons do not appear before 72 hours postfertilization (hpf) and are the last dopaminergic cell group to differentiate. We found that the number of neurons in this cell population continues to increase throughout life proportionally to the growth of the fish. 5-Bromo-2'-deoxyuridine incorporation analysis suggested that this increase is due to continuous neurogenesis and not due to a phenotypic change in already-existing neurons. Finally, expression profiles of several genes (foxg1a, dlx2a, and nr4a2a/b) were different in the POHDA compared with the adjacent suprachiasmatic DA neurons, suggesting that POHDA neurons develop as a distinct DA cell population in the preoptic area. This study offers some insights into the regional identity of the preoptic area and provides the first bases for future functional genetic studies on the development of DA neurons controlling anterior pituitary functions.

The brain reward circuitry plays a key role in emotional and motivational behaviors, and its dysfunction underlies neuropsychiatric disorders such as schizophrenia, depression and drug addiction. Here, we characterized the neuronal activity pattern induced by acute amphetamine administration and during drug-seeking behavior in the zebrafish, and demonstrate the existence of conserved underlying brain circuitry. Combining quantitative analyses of cfos expression with neuronal subtype-specific markers at single-cell resolution, we show that acute d-amphetamine administration leads to both increased neuronal activation and the recruitment of neurons in the medial (Dm) and the lateral (Dl) domains of the adult zebrafish pallium, which contain homologous structures to the mammalian amygdala and hippocampus, respectively. Calbindin-positive and glutamatergic neurons are recruited in Dm, and glutamatergic and γ-aminobutyric acid (GABAergic) neurons in Dl. The drug-activated neurons in Dm and Dl are born at juvenile stage rather than in the embryo or during adulthood. Furthermore, the same territory in Dm is activated during both drug-seeking approach and light avoidance behavior, while these behaviors do not elicit activation in Dl. These data identify the pallial territories involved in acute psychostimulant response and reward formation in the adult zebrafish. They further suggest an evolutionarily conserved function of amygdala-like structures in positive emotions and motivated behavior in zebrafish and mammals.

In contrast with the wealth of data involving bHLH and homeodomain transcription factors in retinal cell type determination, the molecular bases underlying neurotransmitter subtype specification is far less understood. Using both gain and loss of function analyses in Xenopus, we investigated the putative implication of the bHLH factor Ascl1 in this process. We found that in addition to its previously characterized proneural function, Ascl1 also contributes to the specification of the GABAergic phenotype. We showed that it is necessary for retinal GABAergic cell genesis and sufficient in overexpression experiments to bias a subset of retinal precursor cells towards a GABAergic fate. We also analysed the relationships between Ascl1 and a set of other bHLH factors using an in vivo ectopic neurogenic assay. We demonstrated that Ascl1 has unique features as a GABAergic inducer and is epistatic over factors endowed with glutamatergic potentialities such as Neurog2, NeuroD1 or Atoh7. This functional specificity is conferred by the basic DNA binding domain of Ascl1 and involves a specific genetic network, distinct from that underlying its previously demonstrated effects on catecholaminergic differentiation. Our data show that GABAergic inducing activity of Ascl1 requires the direct transcriptional regulation of Ptf1a, providing therefore a new piece of the network governing neurotransmitter subtype specification during retinogenesis.

The receptors of the dopamine neurotransmitter belong to two unrelated classes named D-1 and D-2. For the D-1 receptor class, only two subtypes are found in mammals, the D-1A and D-1B,D- receptors, whereas additional subtypes, named D-1C, D-1D, and D-1X, have been found in other vertebrate species. Here, we analyzed molecular phylogeny, gene synteny, and gene expression pattern of the D-1 receptor subtypes in a large range of vertebrate species, which leads us to propose a new view of the evolution of D-1 dopamine receptor genes. First, we show that D-1C and D-1D receptor sequences are encoded by orthologous genes. Second, the previously identified Cypriniform D-1X sequence is a teleost-specific paralog of the D-1B sequences found in all groups of jawed vertebrates. Third, zebrafish and several sauropsid species possess an additional D-1-like gene, which is likely to form another orthology group of vertebrate ancestral genes, which we propose to name D-1E. Ancestral jawed vertebrates are thus likely to have possessed four classes of D-1 receptor genes-D-1A, D-1B(X), D-1C(D), and D-1E-which arose from large-scale gene duplications. The D-1C receptor gene would have been secondarily lost in the mammalian lineage, whereas the D-1E receptor gene would have been lost independently in several lineages of modern vertebrates. The D-1A receptors are well conserved throughout jawed vertebrates, whereas sauropsid D-1C receptors have rapidly diverged, to the point that they were misidentified as D-1D. The functional significance of the D-1C receptor loss is not known. It is possible that the function may have been substituted with D-1A or D-1B receptors in mammals, following the disappearance of D-1C receptors in these species.

In many teleosts, the stimulatory control of gonadotrope axis by GnRH is opposed by an inhibitory control by dopamine (DA). The functional importance of this inhibitory pathway differs widely from one teleostean species to another. The zebrafish (Danio rerio) is a teleost fish that has become increasingly popular as an experimental vertebrate model. However, the role of DA in the neuroendocrine control of its reproduction has never been studied. Here the authors evaluated in sexually regressed female zebrafish the effects of in vivo treatments with a DA D2 receptor (D2-R) antagonist domperidone, or a GnRH agonist, alone and in combination, on the pituitary level of FSHβ and LHβ transcripts, the gonadosomatic index, and the ovarian histology. Only the double treatment with GnRH agonist and domperidone could induce an increase in the expression of LHβ, in the gonadosomatic index, and a stimulation of ovarian vitellogenesis, indicating that removal of dopaminergic inhibition is required for the stimulatory action of GnRH and reactivation of ovarian function to occur. Using double immunofluorescent staining on pituitary, the authors showed in this species the innervation of LH cells by tyrosine-hydroxylase immunoreactive fibers. Finally, using in situ hybridization and immunofluorescence, the authors showed that the three subtypes of zebrafish DA D2-R (D2a, D2b, and D2c) were expressed in LH-producing cells, suggesting that they all may be involved in mediating this inhibition. These results show for the first time that, in zebrafish, DA has a direct and potent inhibitory action capable of opposing the stimulatory effect of GnRH in the neuroendocrine control of reproduction.

A fundamental duty of any efficient memory system is to prevent long-lasting storage of poorly relevant information. However, little is known about dedicated mechanisms that appropriately trigger production of long-term memory (LTM). We examined the role of Drosophila dopaminergic neurons in the control of LTM formation and found that they act as a switch between two exclusive consolidation pathways leading to LTM or anesthesia-resistant memory (ARM). Blockade, after aversive olfactory conditioning, of three pairs of dopaminergic neurons projecting on mushroom bodies, the olfactory memory center, enhanced ARM, whereas their overactivation conversely impaired ARM. Notably, blockade of these neurons during the intertrial intervals of a spaced training precluded LTM formation. Two pairs of these dopaminergic neurons displayed sustained calcium oscillations in naive flies. Oscillations were weakened by ARM-inducing massed training and were enhanced during LTM formation. Our results indicate that oscillations of two pairs of dopaminergic neurons control ARM levels and gate LTM.

The principal driving force in agricultural research is to increase the yield of food crops. For farming to remain productive, it will be necessary to replenish the nutrients removed or lost from the soil. The objective of this study was to determine the impact of yam-based systems on soil productivity (dry matter production, nutrients recycled or removed, profitability and soil fertility changes). We compared smallholders’ traditional systems (1-year fallow of Andropogonon gayanus -yam rotation; maize-yam rotation) with yam-based systems with legumes (intercropped Aeschynomene histrix with maize-yam rotation; intercropped Mucuna pruriens with maize-yam rotation). The production of dry matter (tubers, shoots), nutrients removed or recycled, and soil properties were significantly improved on yam-based systems with legumes in comparison with traditional systems. Year × Treatment interactions influenced significantly the tuber dry matter production. Site × Treatment and Treatment × Farmer interactions affected significantly nutrients removed or recycled. The amount of nutrients recycled or removed was dependent on the dry matter production that, in turn, depended on soil fertility, rainfall and farmers’ effect.Yam-based systems with legumes brought a higher present value than traditional systems in the first 4 years and appeared attractive for land, labour and cash productivities.

Widely used in biomedical research, zebrafish (Danio rerio) are steadily gaining popularity as a model organism for studying neurobehavioral phenomena.Here, we focus on to the growing spectrum of zebrafish behavioral phenotypes and the 'bigger' biological problems these models help to address.Emphasizing the developing potential of zebrafish as a model organism in biological psychiatry, we discuss several questions related to this field: Do zebrafish have 'emotional'-like behaviors?What are their neural circuits, biomarkers, and ontogenetic origins?And, finally, how can we use this knowledge to build translational bridges to understand human emotions, motivation and personality?Representing a joint effort of several established neurobehavioral laboratories, this article outlines the mounting evidence to support emotionality in zebrafish and other fishes.This conclusion is important to justify the validity of zebrafish 'affective' paradigms and their utility for basic/translational research of normal and pathological behaviors.

Attention-deficit hyperactivity disorder (ADHD) is a developmental disorder characterized by hyperactivity, impulsivity and inattention. It currently affects around 5% of children worldwide and frequently has long-term consequences. However, although there is a significant genetic component to ADHD, relatively few risk genes have been identified and characterized [1-3]. The drug treatments available for the disease are poor with variable efficacy and significant side effects. Furthermore the etiology of ADHD is poorly understood and there are relatively few animal models for the disease. Recently, Arcos-Burgos and colleagues [4] reported evidence for a risk haplotype in the gene encoding Latrophilin 3 (LPHN3), revealed by a linkage scan in a genetic isolate population and subsequent fine mapping. LPHN3 is a potential G protein-coupled receptor with the propensity to moderate cell-cell interaction (adhesion-GPCR). However the physiological function of LPHN3 is not well understood and endogenous ligands have yet to be identified.