Animal cognition: how archer fish learn to down rapidly moving targets

In extremely rapid maneuvers, animals including man can launch ballistic motor patterns that cannot immediately be corrected. Such patterns are difficult to direct at targets that move in three-dimensional space, and it is presently unknown how animals learn to acquire the precision required. Archer fish live in groups and are renowned for their ballistic hunting technique in which they knock down stationary aerial insect prey with a precisely aimed shot of water. Here we report that these fish can learn to release their shots so as to hit prey that moves rapidly at great height, a remarkable accomplishment in which the shooter must take both the target's three-dimensional motion as well as that of its rising shot into account. To successfully perform in the three-dimensional task, training with horizontal motion suffices. Moreover, all archer fish of a group were able to learn the complex sensomotor skill from watching a performing group member, without having to practice. This instance of social learning in a fish is most remarkable as it could imply that observers can "change their viewpoint," mapping the perceived shooting characteristics of a distant team member into angles and target distances that they later must use to hit.     

Coding "what" and "when" in the Archer fish retina

Traditionally, the information content of the neural response is quantified using statistics of the responses relative to stimulus onset time with the assumption that the brain uses onset time to infer stimulus identity. However, stimulus onset time must also be estimated by the brain, making the utility of such an approach questionable. How can stimulus onset be estimated from the neural responses with sufficient accuracy to ensure reliable stimulus identification? We address this question using the framework of colour coding by the archer fish retinal ganglion cell. We found that stimulus identity, “what”, can be estimated from the responses of best single cells with an accuracy comparable to that of the animal''s psychophysical estimation. However, to extract this information, an accurate estimation of stimulus onset is essential. We show that stimulus onset time, “when”, can be estimated using a linear-nonlinear readout mechanism that requires the response of a population of 100 cells. Thus, stimulus onset time can be estimated using a relatively simple readout. However, large nerve cell populations are required to achieve sufficient accuracy.

Authors Summary

In our interaction with the environment we are flooded with a stream of numerous objects and events. Our brain needs to understand the nature of these complex and rich stimuli in order to react. Research has shown ways in which a ‘what’ stimulus was presented can be encoded by the neural responses. However, to understand ‘what was the nature of the stimulus’ the brain needs to know ‘when’ the stimulus was presented. Here, we investigated how the onset of visual stimulus can be signalled by the retina to higher brain regions. We used archer fish as a framework to test the notion that the answer to the question of ‘when’ something has been presented lies within the larger cell population, whereas the answer to the question of ‘what’ has been presented may be found at the single-neuron level. The utility of the archer fish as model animal stems from its remarkable ability to shoot down insects settling on the foliage above the water level, and its ability to distinguish between artificial targets. Thus, the archer fish can provide the fish equivalent of a monkey or a human that can report psychophysical decisions.     

Mechanism of cell-mediated cytotoxicity at the single-cell level: VII. Trigger of the lethal hit event is distinct for NK/K and LDCC effector cells as measured in the two-target conjugate assay

Normal human peripheral blood lymphocytes (PBL) express several in vitro cytotoxic functions, among which are natural killer (NK), antibody-dependent cellular cytotoxicity (ADCC), and lectin-dependent cellular cytotoxicity (LDCC). The relationship of these various cytotoxic functions and the identity of cells involved has been a subject of controversy. Recently it was reported that NK and K for ADCC can be mediated by the same cell, suggesting that they constitute in large part a single subpopulation with multiple cytotoxic functions. The ability of this NK/K effector cell to mediate LDCC was examined here using the two target conjugate assay. The effector cells were Ficoll-Hypaque PBL or LGL-enriched fractions. The targets used were K562 or MOLT for NK, RAJI coated with antibody for ADCC, and RAJI coated with PHA or Con A or modified by NaIO₄ for LDCC. In the two-target conjugate assay, one of the targets is fluorescein labeled for identification. The results show that (a) LDCC copurifies with NK/K and is enriched in the LGL fraction, as measured in both the ⁵¹Cr-release assay and the single-cell assay for cytotoxicity; (b) single effector cells simultaneously bind to NK or ADCC and LDCC targets, revealing that single cells bear binding receptors for all targets; and (c) single lymphocytes were not able to kill both bound NK/K and LDCC targets. However, significant two-target killing was obtained when both targets were NK targets, ADCC targets, LDCC targets, or one NK and one ADCC target. These results demonstrate that the NK and LDCC effector cells are distinct subpopulations copurified in the LGL fraction. In addition, the results show that lectin is unable to trigger globally an NK effector cell to mediate cytotoxicity against a bound NK insensitive target. Thus, although both NK and LDCC effector cells are present in the LGL fraction and can bind to both types of targets, the trigger of the lethal hit event is the function of specialized effector cells.     

Prey catching in the archer fish: does the fish use a learned correction for refraction?

An answer to the question of how the archer fish hits an aerial insect, despite the refraction of light at the surface of the water has not yet been found. The aims of the present studies are to find out: (1) whether the fish applies a learned correction with the virtual image as a point of reference; (2) whether deprivation of practice in squirting affects performance. For the first aim the accuracy of squirts was measured in 30 subjects. Contrary to suggestions from the literature, elevation failures were prominent but the frequencies of over- and under-squirting did not differ, which does not support the idea that the fishes applied a learned correction for refraction by using feedback from the efficacy of squirts. For the second aim, five experimental subjects were deprived of practice, whereas six control subjects got daily practice, during 6 months. The only significant difference, found thereafter, was that during the first session experimental subjects aimed more often before squirting than control subjects did, but hitting was not affected. A number of subjects developed abnormal mandibles which inevitably led to squirting too high. Our findings do not support the hypothesis that the archer fish uses learned corrections for refraction.     

The Effect of Visual Features on Jumping Spider Movements Across Gaps

Our objective was to determine whether an animal’s decisions to cross inhospitable open space are influenced by the visual characteristics of targets it can see across the space. We studied jumping spiders (Salticidae) in the genus Phidippus. We considered the effect of target size (short vs. tall) and distance (close vs. distant) in no-choice experiments. How often spiders approached close targets, regardless of target size, was not significantly different from how often they approached tall, distant targets, but they approached close targets of either size significantly more often than short, distant targets. When presented simultaneously with short, close and tall, distant targets the spiders’ choices did not differ significantly from random. We also tested for the effects of the contrast of targets with their background and found that the spiders crossed open space to reach green, but not white, targets, regardless of background. Finally, spiders were more likely to approach a green grass-like target rather than a target composed of geometric shapes. We conclude that target size, distance and appearance all influence the spiders’ willingness to cross open space.     

Effect of salinity on the refractive index of water: considerations for archer fish aerial vision

The change in the refractive index between fresh and salt water was shown to cause a small shift in the apparent image location of an aerial object viewed from below the water's surface. The impact of this shift may be nominal with respect to archer fish Toxotes spp. spitting accuracy as its magnitude in the direction perpendicular to the spit is close to the width of the spit itself.     

Factors Associated with Shooting Accuracy and Wounding Rate of Four Managed Wild Deer Species in the UK,Based on Anonymous Field Records from Deer Stalkers

The amount of wounding during routine culling is an important factor in the welfare of wild deer. Little information exists on factors determining shooting accuracy and wounding rates under field conditions in the UK. In this study, 102 anonymous stalkers collected data on the outcomes and circumstances of 2281 shots. Using hot-deck imputation and generalised linear mixed modelling, we related the probability that a shot hit its target, and the probability that the shot killed the deer if it was hit, to 28 variables describing the circumstances of the shot. Overall, 96% of deer were hit, of which 93% were killed outright. A reduced probability of hitting the target was associated with an uncomfortable firing position, too little time available, shooting off elbows or freehand, taking the head or upper neck as point of aim, a heavily obscured target, a distant target, shooting at females, lack of shooting practice and a basic (or no) stalker qualification. An increase in the likelihood of wounding was associated with an uncomfortable firing position, shooting with insufficient time, a distant target (only when time was not sufficient), a bullet weight below 75 grains, a target concealed in thicket or on the move and an area rarely stalked. To maximise stalking success and deer welfare, we recommend that stalkers ensure a comfortable firing position, use a gun rest, aim at the chest, use bullets heavier than 75 grains, avoid taking a rushed shot, shoot a distant animal only if there is plenty of time, fire only when the target is stationary, avoid shooting at an obscured animal, take care when the ground is unfamiliar, and do shooting practice at least once a month. The high miss rate of basic-level stalkers suggests that training should include additional firing practice under realistic shooting conditions.     

Mechanism of defective NK cell activity in patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex. II. Normal antibody-dependent cellular cytotoxicity (ADCC) mediated by effector cells defective in natural killer (NK) cytotoxicity

Our studies and other investigations have shown that NK effector cells can also mediate antibody-dependent cellular cytotoxicity (ADCC) through the use of the Fc gamma receptor on the NK cell membrane. Peripheral blood lymphocytes (PBL) derived from patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex exhibit a poor NK activity due to a defective "trigger" required for activation in the lethal hit stage of the NK lytic pathway. Consequently, it was important to delineate whether the defect in AIDS NK cells affected the ADCC function. By using the 51Cr-release assay, the ADCC cytotoxic activity of AIDS PBL was found to be within the normal range, despite the absence of significant NK activity. Several experiments corroborated that the same effector cells mediate both NK CMC and ADCC. Depletion of Fc gamma R-bearing cells resulted in elimination of both the ADCC and NK cytotoxic functions. Single cell analyses, using one- and two-target cell conjugates, revealed that the frequency of ADCC effector:target conjugates and the frequency of killer cells from AIDS PBL were comparable to the frequencies seen in the normal controls. However, when mixtures of NK and ADCC targets were used to form mixed two-target conjugates, the AIDS effector cells lysed only the bound ADCC target, whereas the normal effector cells lysed both the bound NK and ADCC targets. These results demonstrate clearly that the same NK/K effector cells from AIDS PBL, defective in NK activity, are not impaired in mediating ADCC activity. These findings were supported by the demonstration that AIDS PBL stimulated with ADCC targets, but not with NK targets, released NK cytotoxic factors, postulated mediators of the NK CMC reaction. These findings indicate that the NK/K cells in AIDS are triggered normally for ADCC activity but are not triggered for NK activity. Furthermore, the results indicate that the lytic machinery is not impaired in the AIDS NK/K cells.     

Archer fish learn to compensate for complex optical distortions to determine the absolute size of their aerial prey

Many animals, including humans, can visually judge the absolute size of objects regardless of changes in viewing distance and thus despite the resulting dramatic differences in the size of the actual retinal images. For animals that have to judge the size of aerial objects from underwater views, this can be a formidable problem; our calculations show that considerable and strongly viewpoint-dependent corrections are needed to compensate for the effects of light refraction. Archer fish face these optical difficulties because they have to shoot down aerial insects over a wide range of horizontal and vertical distances. We show here that these fish can learn to acquire size constancy with remarkable precision and are thus fully capable of taking complex viewpoint dependency into account. Moreover, we demonstrate that archer fish solve the problem not by interpolating within a set of stored views and distances but by learning the laws that connect apparent size with the fish's relative position to the target. This enables the fish to readily judge the absolute sizes of objects from completely novel views.     

A Superfamily of DNA Transposons Targeting Multicopy Small RNA Genes

Target-specific integration of transposable elements for multicopy genes, such as ribosomal RNA and small nuclear RNA (snRNA) genes, is of great interest because of the relatively harmless nature, stable inheritance and possible application for targeted gene delivery of target-specific transposable elements. To date, such strict target specificity has been observed only among non-LTR retrotransposons. We here report a new superfamily of sequence-specific DNA transposons, designated Dada. Dada encodes a DDE-type transposase that shows a distant similarity to transposases encoded by eukaryotic MuDR, hAT, P and Kolobok transposons, as well as the prokaryotic IS256 insertion element. Dada generates 6–7 bp target site duplications upon insertion. One family of Dada DNA transposons targets a specific site inside the U6 snRNA genes and are found in various fish species, water flea, oyster and polycheate worm. Other target sequences of the Dada transposons are U1 snRNA genes and different tRNA genes. The targets are well conserved in multicopy genes, indicating that copy number and sequence conservation are the primary constraints on the target choice of Dada transposons. Dada also opens a new frontier for target-specific gene delivery application.     

Prediction of Drug-Target Interactions for Drug Repositioning Only Based on Genomic Expression Similarity

Small drug molecules usually bind to multiple protein targets or even unintended off-targets. Such drug promiscuity has often led to unwanted or unexplained drug reactions, resulting in side effects or drug repositioning opportunities. So it is always an important issue in pharmacology to identify potential drug-target interactions (DTI). However, DTI discovery by experiment remains a challenging task, due to high expense of time and resources. Many computational methods are therefore developed to predict DTI with high throughput biological and clinical data. Here, we initiatively demonstrate that the on-target and off-target effects could be characterized by drug-induced in vitro genomic expression changes, e.g. the data in Connectivity Map (CMap). Thus, unknown ligands of a certain target can be found from the compounds showing high gene-expression similarity to the known ligands. Then to clarify the detailed practice of CMap based DTI prediction, we objectively evaluate how well each target is characterized by CMap. The results suggest that (1) some targets are better characterized than others, so the prediction models specific to these well characterized targets would be more accurate and reliable; (2) in some cases, a family of ligands for the same target tend to interact with common off-targets, which may help increase the efficiency of DTI discovery and explain the mechanisms of complicated drug actions. In the present study, CMap expression similarity is proposed as a novel indicator of drug-target interactions. The detailed strategies of improving data quality by decreasing the batch effect and building prediction models are also effectively established. We believe the success in CMap can be further translated into other public and commercial data of genomic expression, thus increasing research productivity towards valid drug repositioning and minimal side effects.     

ATM regulates target switching to escalating doses of radiation in the intestines

Although stem cells succumbing to reproductive death are assumed to be the single relevant targets in radiation tissue damage, recent studies showed intestinal stem cell damage is conditionally linked to crypt endothelial apoptosis, defining a two-target model. Here we report that when mouse intestines were protected against microvascular apoptosis, radiation switched as the dose escalated to a previously unrecognized crypt stem cell target, activating ceramide synthase-mediated apoptosis to initiate intestinal damage. Whereas ataxia telangiectasia-mutated (ATM) kinase normally represses ceramide synthase, its derepression in Atm(-/-) mice increased crypt stem cell radiosensitivity 3.7-fold without sensitizing the microvascular response. Discovery of this intestinal radiosensitivity mechanism allowed design of an antisense Atm oligonucleotide treatment which phenocopied the Atm(-/-) mouse, reordering ceramide synthase-mediated stem cell death to become the first-line gastrointestinal response of wild-type littermates. These experiments indicate that tissues operate multiple potential targets activated consecutively according to their inherent radiosensitivities that may be reordered therapeutically to control radiation tissue responses.     

Haptic Guidance Needs to Be Intuitive Not Just Informative to Improve Human Motor Accuracy

Humans make both random and systematic errors when reproducing learned movements. Intuitive haptic guidance that assists one to make the movements reduces such errors. Our study examined whether any additional haptic information about the location of the target reduces errors in a position reproduction task, or whether the haptic guidance needs to be assistive to do so. Holding a haptic device, subjects made reaches to visible targets without time constraints. They did so in a no-guidance condition, and in guidance conditions in which the direction of the force with respect to the target differed, but the force scaled with the distance to the target in the same way. We examined whether guidance forces directed towards the target would reduce subjects’ errors in reproducing a prior position to the same extent as do forces rotated by 90 degrees or 180 degrees, as it might because the forces provide the same information in all three cases. Without vision of the arm, both the accuracy and precision were significantly better with guidance directed towards the target than in all other conditions. The errors with rotated guidance did not differ from those without guidance. Not surprisingly, the movements tended to be faster when guidance forces directed the reaches to the target. This study shows that haptic guidance significantly improved motor performance when using it was intuitive, while non-intuitively presented information did not lead to any improvements and seemed to be ignored even in our simple paradigm with static targets and no time constraints.     

Do People “Pop Out”?

The human body is a highly familiar and socially very important object. Does this mean that the human body has a special status with respect to visual attention? In the current paper we tested whether people in natural scenes attract attention and “pop out” or, alternatively, are at least searched for more efficiently than targets of another category (machines). Observers in our study searched a visual array for dynamic or static scenes containing humans amidst scenes containing machines and vice versa. The arrays consisted of 2, 4, 6 or 8 scenes arranged in a circular array, with targets being present or absent. Search times increased with set size for dynamic and static human and machine targets, arguing against pop out. However, search for human targets was more efficient than for machine targets as indicated by shallower search slopes for human targets. Eye tracking further revealed that observers made more first fixations to human than to machine targets and that their on-target fixation durations were shorter for human compared to machine targets. In summary, our results suggest that searching for people in natural scenes is more efficient than searching for other categories even though people do not pop out.     

MSLN (Mesothelin), ANTXR1 (TEM8), and MUC3A are the potent antigenic targets for CAR T cell therapy of gastric adenocarcinoma

Gastric adenocarcinoma is usually diagnosed in late stages, necessitating the use of different therapeutic modalities. Currently, antibody-based therapies have also been approved through with limited clinical efficacy. Reinforcing antibody-based immunotherapy by using chimeric antigen receptor (CAR) T cells may enhance the approach. However, the cells can cause severe on-target and off-tumor toxicities owing to their higher sensitivity to low-level antigen expressions. To address the need for safe and reliable targets, we made a bioinformatics pipeline by which we screened overexpressed genes in the disease for off-tumor sites in many normal tissues. Our inspection showed that MSLN (Mesothelin), ANTXR1 (TEM8), and MUC3A are the probable targets of CAR T cell therapy in gastric adenocarcinoma. The proposed antigenic targets might respond to the need to simultaneously target multiple antigens in a tumor matrix to prevent resistance.     

Effect of archer density on elk pregnancy rates and conception dates

Archery hunting in Oregon has increased dramatically over the past 2 decades. At the same time, spring juvenile to adult female ratios of Rocky Mountain elk (Cervus elaphus) have been declining. This has raised concern that archery seasons may be disrupting elk breeding and contributing to the decline in recruitment. Two mechanisms could contribute to reduced juvenile:female ratios: 1) reduced pregnancy rates, and 2) delayed conception dates because of human disturbance during the rut. We varied the number of archery hunters at the Starkey Experimental Forest and Range over 13 years to evaluate effects of archer density on reproduction of elk. Archer densities were maintained at high densities during 4 years , low densities during 3 years , and no archers during 6 years. We determined pregnancy status, age, kidney fat index (KFI), lactation status, and fetus conception dates for 622 female elk harvested in December. We found pregnancy rate differences of 0.105, 0.080, and 0.021 between high and no archer density years (P = 0.004), high and low archer density years (P = 0.054), and low and no archer density years (P = 0.616), respectively. Conception dates were 4 days later for high archer density compared to low archer density (P = 0.006), but did not differ between high and no archer years (2 days; P = 0.136) or between low and no archer years (2 days; P = 0.108). We compared generalized linear model estimates of pregnancy rates and determined pregnancy rates for 28% of the lactating female elk to be affected by high archer density, whereas archer densities had no significant affect on pregnancy rate estimates for non-lactating females. We found no difference in conception dates among archer densities when comparing model estimates. Our results suggest that archer density and its interaction with nutritional condition of elk influence pregnancy rates of lactating females with low KFI levels; however, the effect of archer density alone does not explain the magnitude of decline in juvenile to female ratios observed in Oregon. © 2012 The Wildlife Society.     

Developmental programmes of growth and survival in early sensory neurons.

In the developing vertebrate nervous system the survival of neurons becomes dependent on the supply of a neurotrophic factor from their targets when their axons reach these targets. To determine how the onset of neurotrophic factor dependency is coordinated with the arrival of axons in the target field, we have studied the growth and survival of four populations of cranial sensory neurons whose axons have markedly different distances to grow to reach their targets. Axonal growth rate both in vivo and in vitro is related to target distance; neurons with more distant targets grow faster. The onset trophic factor dependency in culture is also related to target distance; neurons with more distant targets survive longer before becoming trophic factor dependent. These data suggest that programmes of growth and survival in early neurons play an important role in coordinating the timing of trophic interactions in the developing nervous system.     

Multivariate assessments of activity-related skeletal changes: Interpreting Bell Beaker specialized male archery and social organization in Central Europe

Objectives

The Bell Beaker period witnessed the rise of individual inhumations with “wealthy” burial contexts containing archery-related grave goods, leading archaeologists to label the individuals in these tombs as “archers.” This study looks to (1) compare the skeletons from male “archer” burials with those from male “non-archer” burials—those not having archery-related grave goods—in order to assess a possible link between burial context and physical activity, and (2) apply a biomechanics profile to evaluate whether the individuals associated with these “archer” burials practiced specialized archer activity.

Materials and Methods

The corpus (males only) included 46 “archers” and 40 “non-archers” from Bell Beaker individual inhumations. Osteological data included measurements, scores of entheseal changes, and a diagnosis of certain pathologies. Data analyses involved visual observations, hypothesis tests, dimension reduction, and MANOVA, with approaches aimed at exploring the treatment of data missingness.

Results

Measurement data revealed no differences between the two groups. Evaluations of entheseal changes found that “non-archers” had consistently more instances of bone surface modifications than “archers.” Individual assessments of specialized archer occupation identified 11 possible specialized archers.

Discussion

These findings indicate a possible labor differentiation represented through the presence of a probably prestigious “archer” burial context. This suggests a link between grave good presence and labor, but not between a Bell Beaker archery occupation and an “archer” burial context. Data analyses support the application of biomechanics to osteological analyses in order to assess specialized activity on the skeleton.