Distinct Circuits for the Formation and Retrieval of an Imprinted Olfactory Memory.

Memories formed early in life are particularly stable and influential, representing privileged experiences that shape enduring behaviors. We show that exposing newly hatched C. elegans to pathogenic bacteria results in persistent aversion to those bacterial odors, whereas adult exposure generates only transient aversive memory. Long-lasting imprinted aversion has a critical period in the first larval stage and is specific to the experienced pathogen. Distinct groups of neurons are required during formation (AIB, RIM) and retrieval (AIY, RIA) of the imprinted memory. RIM synthesizes the neuromodulator tyramine, which is required in the L1 stage for learning. AIY memory retrieval neurons sense tyramine via the SER-2 receptor, which is essential for imprinted, but not for adult-learned, aversion. Odor responses in several neurons, most notably RIA, are altered in imprinted animals. These findings provide insight into neuronal substrates of different forms of memory, and lay a foundation for further understanding of early learning.

Behavioral evidence for geomagnetic imprinting and transgenerational inheritance in fruit flies.

Certain long-distance migratory animals, such as salmon and sea turtles, are thought to imprint on the magnetic field of their natal area and to use this information to help them return as adults. Despite a growing body of indirect support for such imprinting, direct experimental evidence thereof remains elusive. Here, using the fruit fly as a magnetoreceptive model organism, we demonstrate that exposure to a specific geographic magnetic field during a critical period of early development affected responses to a matching magnetic field gradient later in life. Specifically, hungry flies that had imprinted on a specific magnetic field from 1 of 3 widely separated geographic locations responded to the imprinted field, but not other magnetic fields, by moving downward, a geotactic behavior associated with foraging. This same behavior occurred spontaneously in the progeny of the next generation: female progeny moved downward in response to the field on which their parents had imprinted, whereas male progeny did so only in the presence of these females. These results represent experimental evidence that organisms can learn and remember a magnetic field to which they were exposed during a critical period of development. Although the function of the behavior is not known, one possibility is that imprinting on the magnetic field of a natal area assists flies and their offspring in recognizing locations likely to be favorable for foraging and reproduction.

Imprinting on time-structured acoustic stimuli in ducklings.

Filial imprinting is a dedicated learning process that lacks explicit reinforcement. The phenomenon itself is narrowly heritably canalized, but its content, the representation of the parental object, reflects the circumstances of the newborn. Imprinting has recently been shown to be even more subtle and complex than previously envisaged, since ducklings and chicks are now known to select and represent for later generalization abstract conceptual properties of the objects they perceive as neonates, including movement pattern, heterogeneity and inter-component relationships of same or different. Here, we investigate day-old Mallard (Anas platyrhynchos) ducklings' bias towards imprinting on acoustic stimuli made from mallards' vocalizations as opposed to white noise, whether they imprint on the temporal structure of brief acoustic stimuli of either kind, and whether they generalize timing information across the two sounds. Our data are consistent with a strong innate preference for natural sounds, but do not reliably establish sensitivity to temporal relations. This fits with the view that imprinting includes the establishment of representations of both primary percepts and selective abstract properties of their early perceptual input, meshing together genetically transmitted prior pre-dispositions with active selection and processing of the perceptual input.

Steps towards a computational ethology: an automatized, interactive setup to investigate filial imprinting and biological predispositions.

Soon after hatching, the young of precocial species, such as domestic chicks or ducklings, learn to recognize their social partner by simply being exposed to it (imprinting process). Even artificial objects or stimuli displayed on monitor screens can effectively trigger filial imprinting, though learning is canalized by spontaneous preferences for animacy signals, such as certain kinds of motion or a face-like appearance. Imprinting is used as a behavioural paradigm for studies on memory formation, early learning and predispositions, as well as number and space cognition, and brain asymmetries. Here, we present an automatized setup to expose and/or test animals for a variety of imprinting experiments. The setup consists of a cage with two high-frequency screens at the opposite ends where stimuli are shown. Provided with a camera covering the whole space of the cage, the behaviour of the animal is recorded continuously. A graphic user interface implemented in Matlab allows a custom configuration of the experimental protocol, that together with Psychtoolbox drives the presentation of images on the screens, with accurate time scheduling and a highly precise framerate. The setup can be implemented into a complete workflow to analyse behaviour in a fully automatized way by combining Matlab (and Psychtoolbox) to control the monitor screens and stimuli, DeepLabCut to track animals' behaviour, Python (and R) to extract data and perform statistical analyses. The automated setup allows neuro-behavioural scientists to perform standardized protocols during their experiments, with faster data collection and analyses, and reproducible results.

Role of sexual imprinting in assortative mating and premating isolation in Darwin's finches.

Global biodiversity is being degraded at an unprecedented rate, so it is important to preserve the potential for future speciation. Providing for the future requires understanding speciation as a contemporary ecological process. Phylogenetically young adaptive radiations are a good choice for detailed study because diversification is ongoing. A key question is how incipient species become reproductively isolated from each other. Barriers to gene exchange have been investigated experimentally in the laboratory and in the field, but little information exists from the quantitative study of mating patterns in nature. Although the degree to which genetic variation underlying mate-preference learning is unknown, we provide evidence that two species of Darwin's finches imprint on morphological cues of their parents and mate assortatively. Statistical evidence of presumed imprinting is stronger for sons than for daughters and is stronger for imprinting on fathers than on mothers. In combination, morphology and species-specific song learned from the father constitute a barrier to interbreeding. The barrier becomes stronger the more the species diverge morphologically and ecologically. It occasionally breaks down, and the species hybridize. Hybridization is most likely to happen when species are similar to each other in adaptive morphological traits, e.g., body size and beak size and shape. Hybridization can lead to the formation of a new species reproductively isolated from the parental species as a result of sexual imprinting. Conservation of sufficiently diverse natural habitat is needed to sustain a large sample of extant biota and preserve the potential for future speciation.

What can a worm learn in a bacteria-rich habitat?

With a nervous system that has only a few hundred neurons, Caenorhabditis elegans was initially not regarded as a model for studies on learning. However, the collective effort of the C. elegans field in the past several decades has shown that the worm displays plasticity in its behavioral response to a wide range of sensory cues in the environment. As a bacteria-feeding worm, C. elegans is highly adaptive to the bacteria enriched in its habitat, especially those that are pathogenic and pose a threat to survival. It uses several common forms of behavioral plasticity that last for different amounts of time, including imprinting and adult-stage associative learning, to modulate its interactions with pathogenic bacteria. Probing the molecular, cellular and circuit mechanisms underlying these forms of experience-dependent plasticity has identified signaling pathways and regulatory insights that are conserved in more complex animals.

Combined predisposed preferences for colour and biological motion make robust development of social attachment through imprinting.

To study how predisposed preferences shape the formation of social attachment through imprinting, newly hatched domestic chicks (Gallus gallus domesticus) were simultaneously exposed to two animations composed of comparable light points in different colours (red and yellow), one for a walking motion and another for a linear motion. When a walking animation in red was combined with a linear one in yellow, chicks formed a learned preference for the former that represented biological motion (BM). When the motion-colour association was swapped, chicks failed to form a preference for a walking in yellow, indicating a bias to a specific association of motion and colour. Accordingly, experiments using realistic walking chicken videos revealed a preference for a red video over a yellow one, when the whole body or the head was coloured. On the other hand, when the BM preference had been pre-induced using an artefact moving rigidly (non-BM), a clear preference for a yellow walking animation emerged after training by the swapped association. Even if the first-seen moving object was a nonbiological artefact such as the toy, the visual experience would induce a predisposed BM preference, making chicks selectively memorize the object with natural features. Imprinting causes a rapid inflow of thyroid hormone in the telencephalon leading to the induction of the BM preference, which would make the robust formation of social attachment selectively to the BM-associated object such as the mother hen.

Cross-continental test of natal philopatry and habitat-imprinting hypotheses to explain host specificity in an obligate brood parasite.

Several hypotheses have been proposed to explain the mechanisms responsible for maintenance of host-specific gentes in the common cuckoo (Cuculus canorus). Some of them expect that when adult cuckoos return to lay their eggs to their natal site (natal philopatry hypothesis) or habitat in which they were reared (habitat-imprinting hypothesis), there is a higher probability of finding nests of the host species by which they were reared. Since published evidence is ambiguous, we here evaluate the natal philopatry and habitat-imprinting hypotheses using information on habitat homogeneity and cross-continental long-term ringing data. We found no evidence for the natal philopatry hypothesis-instead of returning to their natal site, juvenile cuckoos exhibited longer dispersal movements than adults, and the difference was even larger in comparison with a wide array of cuckoo host species. On the contrary, we found support for the habitat-imprinting hypothesis-juvenile cuckoos followed similar levels of natal habitat homogeneity at 5- and 25-km scale when returning to breed in the next years. Our results suggest that preference for the particular habitat structures may help cuckoos to find appropriate hosts.

The cow-calf relationship: from maternal responsiveness to the maternal bond and the possibilities for fostering.

The relationship between the cow and calf develops over time after birth. The behavioural mechanisms underlying its development are important and comparisons with other species may increase our understanding. In nature the cow will separate herself from the herd to give birth and then the cow-calf relationship will develop with the ability to recognise each other. While twinning levels are low in cows, they do rear their twin calves. If the calf is lost at or after birth the cow can be responsive towards other calves and in specific circumstances the cow can develop a maternal bond with an alien calf, i.e. foster. In this Research Reflection a distinction is made between the development of, on the one hand, maternal responsiveness (the tendency of the cow to care for a calf which occurs before birth) and, on the other hand, the development of the maternal-filial bond or relationship which is reciprocal, occurs after birth and is characterised by the ability to discriminate the mother's own calf from alien calves. These processes can overlap and the relationship between cow and calf in this 'hider' species is more plastic than in some other mammals. For example, a cow might form an attachment with an alien calf before she gives birth. After the cow has given birth the loss of her own calf may result in the state of maternal responsiveness being maintained, such that developing a maternal bond with one or several appropriate alien calves is possible. Viable fostering techniques are possible. If a maternal relationship to the mother's own calf has developed then fostering will be more difficult. If the cow's relationship with her own calf is not exclusive, and she is in a state of maternal responsiveness then fostering of calves of an appropriate age and status can be achieved.

Behavioural and neuronal basis of olfactory imprinting and kin recognition in larval fish.

Imprinting is a specific form of long-term memory of a cue acquired during a sensitive phase of development. To ensure that organisms memorize the right cue, the learning process must happen during a specific short time period, mostly soon after hatching, which should end before irrelevant or misleading signals are encountered. A well-known case of olfactory imprinting in the aquatic environment is that of the anadromous Atlantic and Pacific salmon, which prefer the olfactory cues of natal rivers to which they return after migrating several years in the open ocean. Recent research has shown that olfactory imprinting and olfactory guided navigation in the marine realm are far more common than previously assumed. Here, we present evidence for the involvement of olfactory imprinting in the navigation behaviour of coral reef fish, which prefer their home reef odour over that of other reefs. Two main olfactory imprinting processes can be differentiated: (1) imprinting on environmental cues and (2) imprinting on chemical compounds released by kin, which is based on genetic relatedness among conspecifics. While the first process allows for plasticity, so that organisms can imprint on a variety of chemical signals, the latter seems to be restricted to specific genetically determined kin signals. We focus on the second, elucidating the behavioural and neuronal basis of the imprinting process on kin cues using larval zebrafish (Danio rerio) as a model. Our data suggest that the process of imprinting is not confined to the central nervous system but also triggers some changes in the olfactory epithelium.

Ducklings imprint on chromatic heterogeneity.

Avian filial imprinting is a rapid form of learning occurring just after hatching in precocial bird species. The acquired imprint on either or both parents goes on to affect the young bird's survival and social behaviour later in life (Bateson in Biol Rev 41:177-217, 1966). The imprinting mechanism is specialized but flexible, and causes the hatchling to develop high-fidelity recognition and attraction to any moving stimulus of suitable size seen during a predefined sensitive period. It has been observed (Martinho and Kacelnik in Science 353:286-288, 2016; Versace et al. in Anim Cogn 20:521-529, 2017) that in addition to visual and acoustic sensory inputs, imprinting may incorporate informational rules or abstract concepts. Here we report a study of mallard ducklings (Anas platyrhynchos domesticus) undergoing imprinting on the chromatic heterogeneity of stimuli, with a focus on how this may be transferred to novel objects. Ducklings were exposed to a series of chromatically heterogeneous or homogeneous stimuli and tested for preference between two novel stimuli, one heterogeneous and the other homogeneous. Exposure to heterogeneity significantly enhanced preference for novel heterogeneous stimuli, relative to ducklings exposed to homogeneous stimuli or unexposed controls. These findings support the view that imprinting does not rely solely on exemplars, or snapshot-like representations of visual input, but that instead young precocial animals form complex multidimensional representations of the target object, involving abstract properties, either at the time of learning, or later, through generalization from the learnt exemplars.

Sensory mechanisms of natal stream imprinting and homing in Oncorhynchus spp.

Juvenile Oncorhynchus spp. can memorise their natal stream during downstream migration; juveniles migrate to feed during their growth phase and then they migrate long distances from their feeding habitat to their natal stream to reproduce as adults. Two different sensory mechanisms, olfaction and navigation, are involved in the imprinting and homing processes during short-distance migration within the natal stream and long-distance migration in open water, respectively. Here, olfactory functions are reviewed from both neurophysiological studies on the olfactory discrimination ability of natal stream odours and neuroendocrinological studies on the hormonal controlling mechanisms of olfactory memory formation and retrieval in the brain. These studies revealed that the long-term stability of dissolved free amino-acid composition in the natal stream is crucial for olfactory imprinting and homing. Additionally, the brain-pituitary-thyroid and brain-pituitary-gonadal hormones play important roles in olfactory memory formation and retrieval, respectively. Navigation functions were reviewed from physiological biotelemetry techniques with sensory interference experiments during the homing migration of anadromous and lacustrine Oncorhynchus spp. The experiments demonstrated that Oncorhynchus spp. used compass navigation mechanisms in the open water. These findings are discussed in relation to the sensory mechanisms involved in natal stream imprinting and homing in Oncorhynchus spp.

Wnt-2b in the intermediate hyperpallium apicale of the telencephalon is critical for the thyroid hormone-mediated opening of the sensitive period for filial imprinting in domestic chicks (Gallus gallus domesticus).

Filial imprinting is the behavior observed in chicks during the sensitive or critical period of the first 2-3 days after hatching; however, after this period they cannot be imprinted when raised in darkness. Our previous study showed that temporal augmentation of the endogenous thyroid hormone 3,5,3'-triiodothyronine (T3) in the telencephalon, by imprinting training, starts the sensitive period just after hatching. Intravenous injection of T3 enables imprinting of chicks on days 4 or 6 post-hatching, even when the sensitive period has ended. However, the molecular mechanism of how T3 acts as a determinant of the sensitive period is unknown. Here, we show that Wnt-2b mRNA level is increased in the T3-injected telencephalon of 4-day old chicks. Pharmacological inhibition of Wnt signaling in the intermediate hyperpallium apicale (IMHA), which is the caudal area of the telencephalon, blocked the recovery of the sensitive period following T3 injection. In addition, injection of recombinant Wnt-2b protein into the IMHA helped chicks recover the sensitive period without the injection of T3. Lastly, we showed Wnt signaling to be involved in imprinting via the IMHA region on day 1 during the sensitive period. These results indicate that Wnt signaling plays a critical role in the opening of the sensitive period downstream of T3.

Feeding Imprinting: The Extreme Test Case of Premature Infants Born With Very Low Birth Weight.

Feeding imprinting, considered a survival-enabling process, is not well understood. Infants born very preterm, who first feed passively, are an effective model for studying feeding imprinting. Retrospective analysis of neonatal intensive care unit (NICU) records of 255 infants (Mgestational age  = 29.98 ± 1.64) enabled exploring the notion that direct breastfeeding (DBF) during NICU stay leads to consumption of more mother's milk and earlier NICU discharge. Results showed that DBF before the first bottle feeding is related to shorter transition into oral feeding, a younger age of full oral feeding accomplishment and earlier discharge. Furthermore, the number of DBF meals before first bottle feeding predicts more maternal milk consumption and improved NICU outcomes. Improved performance in response to initial exposure to DBF at the age of budding feeding abilities supports a feeding imprinting hypothesis.

Molecular characterization and gene expression of synaptosome-associated protein-25 (SNAP-25) in the brain during both seaward and homeward migrations of chum salmon Oncorhynchus keta.

It is generally accepted that information about some of the odorants in the natal streams of anadromous Pacific salmon (Genus Oncorhynchus) is imprinted during their seaward migration, and that anadromous Pacific salmon use olfaction to identify their natal streams during the homeward migration. However, little is known about the molecular mechanisms of the various pre-synaptic functions that are important for olfactory imprinting and memory retrieval in the salmon brain. Synaptosome-associated protein-25 (SNAP-25) mediates pre-synaptic vesicle exocytosis and regulates synaptic transmission and neuronal plasticity. Despite the importance of synaptic plasticity for memorization, the expression of SNAP-25 in the salmon brain is not well understood. In this study, snap25 expression was detected in chum salmon (O. keta) brains using molecular biological techniques. Two cDNAs encoding salmon SNAP-25 were isolated and sequenced (SNAP-25a and SNAP-25b). These cDNAs encoded proteins with 204 amino acid residues, which showed marked homology with each other (97%). The protein and nucleotide sequences demonstrated a high level of homology between salmon SNAP-25s and those of other teleost species. By quantitative PCR, the expression of snap25a and snap25b was detected in all regions of the salmon brain, especially in the telencephalon. The expression levels of snap25a in the olfactory blub were higher during seaward migration than in upriver and post-upriver migrations, reflecting synaptogenesis in the olfactory nervous system, and snap25b in the telencephalon was increased during upriver period. Our results indicated that snap25s gene is involved in synaptic plasticity for olfactory imprinting and/or olfactory memory retrieval in Pacific salmon.

Spontaneous generalization of abstract multimodal patterns in young domestic chicks.

From the early stages of life, learning the regularities associated with specific objects is crucial for making sense of experiences. Through filial imprinting, young precocial birds quickly learn the features of their social partners by mere exposure. It is not clear though to what extent chicks can extract abstract patterns of the visual and acoustic stimuli present in the imprinting object, and how they combine them. To investigate this issue, we exposed chicks (Gallus gallus) to three days of visual and acoustic imprinting, using either patterns with two identical items or patterns with two different items, presented visually, acoustically or in both modalities. Next, chicks were given a choice between the familiar and the unfamiliar pattern, present in either the multimodal, visual or acoustic modality. The responses to the novel stimuli were affected by their imprinting experience, and the effect was stronger for chicks imprinted with multimodal patterns than for the other groups. Interestingly, males and females adopted a different strategy, with males more attracted by unfamiliar patterns and females more attracted by familiar patterns. Our data show that chicks can generalize abstract patterns by mere exposure through filial imprinting and that multimodal stimulation is more effective than unimodal stimulation for pattern learning.

Comparative approaches to same/different abstract-concept learning.

Martinho and Kacelnik (2016) imprinted newly hatched ducklings (Anas platyrhynchos domestica) with a moving pair of either same or different objects, and following only one session, the ducklings accurately transferred the same/different relationship to novel object pairs that maintained the training relationship. This rapid learning and transfer of the concepts same and different far outstrips the more gradual learning of these basic concepts by animals in associative-learning tasks in which reinforcement is given for correct responses.

Natal habitat imprinting counteracts the diversifying effects of phenotype-dependent dispersal in a spatially structured population.

BACKGROUND: Habitat selection may have profound evolutionary consequences, but they strongly depend on the underlying preference mechanism, including genetically-determined, natal habitat and phenotype-dependent preferences. It is known that different mechanisms may operate at the same time, yet their relative contribution to population differentiation remains largely unexplored empirically mainly because of the difficulty of finding suitable study systems. Here, we investigate the role of early experience and genetic background in determining the outcome of settlement by pied flycatchers (Ficedula hypoleuca) breeding in two habitat patches between which dispersal and subsequent reproductive performance is influenced by phenotype (body size). For this, we conducted a cross-fostering experiment in a two-patch system: an oakwood and a conifer plantation separated by only 1 km. RESULTS: Experimental birds mostly returned to breed in the forest patch where they were raised, whether it was that of their genetic or their foster parents, indicating that decisions on where to settle are determined by individuals' experience in their natal site, rather than by their genetic background. Nevertheless, nearly a third (27.6 %) moved away from the rearing habitat and, as previously observed in unmanipulated individuals, dispersal between habitats was phenotype-dependent. Pied flycatchers breeding in the oak and the pine forests are differentiated by body size, and analyses of genetic variation at microsatellite loci now provide evidence of subtle genetic differentiation between the two populations. This suggests that phenotype-dependent dispersal may contribute to population structure despite the short distance and widespread exchange of birds between the study plots. CONCLUSIONS: Taken together, the current and previous findings that pied flycatchers do not always settle in the habitat to which they are best suited suggest that their strong tendency to return to the natal patch regardless of their body size might lead to maladaptive settlement decisions and thus constrain the potential of phenotype-dependent dispersal to promote microgeographic adaptation.

The development of newborn object recognition in fast and slow visual worlds.

Object recognition is central to perception and cognition. Yet relatively little is known about the environmental factors that cause invariant object recognition to emerge in the newborn brain. Is this ability a hardwired property of vision? Or does the development of invariant object recognition require experience with a particular kind of visual environment? Here, we used a high-throughput controlled-rearing method to examine whether newborn chicks (Gallus gallus) require visual experience with slowly changing objects to develop invariant object recognition abilities. When newborn chicks were raised with a slowly rotating virtual object, the chicks built invariant object representations that generalized across novel viewpoints and rotation speeds. In contrast, when newborn chicks were raised with a virtual object that rotated more quickly, the chicks built viewpoint-specific object representations that failed to generalize to novel viewpoints and rotation speeds. Moreover, there was a direct relationship between the speed of the object and the amount of invariance in the chick's object representation. Thus, visual experience with slowly changing objects plays a critical role in the development of invariant object recognition. These results indicate that invariant object recognition is not a hardwired property of vision, but is learned rapidly when newborns encounter a slowly changing visual world.

Imprinting can cause a maladaptive preference for infectious conspecifics.

Recognizing and associating with specific individuals, such as conspecifics or kin, brings many benefits. One mechanism underlying such recognition is imprinting: the long-term memory of cues encountered during development. Typically, juveniles imprint on cues of nearby individuals and may later associate with phenotypes matching their 'recognition template'. However, phenotype matching could lead to maladaptive social decisions if, for instance, individuals imprint on the cues of conspecifics infected with directly transmitted diseases. To investigate the role of imprinting in the sensory ecology of disease transmission, we exposed juvenile guppies,Poecilia reticulata, to the cues of healthy conspecifics, or to those experiencing disease caused by the directly transmitted parasite Gyrodactylus turnbulli In a dichotomous choice test, adult 'disease-imprinted' guppies preferred to associate with the chemical cues of G. turnbulli-infected conspecifics, whereas 'healthy-imprinted' guppies preferred to associate with cues of uninfected conspecifics. These responses were only observed when stimulus fish were in late infection, suggesting imprinted fish responded to cues of disease, but not of infection alone. We discuss how maladaptive imprinting may promote disease transmission in natural populations of a social host.