Thyroid Hormone Sensitizes the Imprinting-Associated Induction of Biological Motion Preference in Domestic Chicks

Filial imprinting is associated with induction of predisposed preference to animations that bear visual features of Johansson's biological motion (BM), and the induction is limited to a few days after hatching. As thyroid hormone (3,5,3′-triiodothyronine, T3) plays a critical role in determining the sensitive period of imprinting, we examined if exogenously applied T3 (or iopanoic acid, IOP; a selective inhibitor for converting enzymes) could also sensitize (or desensitize) the BM induction. Chicks were trained by using a non-BM stimulus (rotating red toy) according to a conventional imprinting procedure. Trained chicks were tested for preference to a point-light BM animation (walking chick) over a non-BM animation (linear motion), and for the preference for the familiarized stimulus (red toy) over an unfamiliar one (yellow toy). In 1-day chicks, those injected with IOP showed significantly lower scores than controls on both BM and imprinting tests. In 4-days chicks, those injected with T3 showed higher scores than control, but the difference in BM score was not significant. Imprinting and the accompanying T3 surge may be necessary for the predisposed BM preference to appear in 1-day chicks. Even after the conventional sensitive period is over, exogenous T3 can partly re-sensitize the BM preference as it does imprinting

GABA-A and GABA-B Receptors in Filial Imprinting Linked With Opening and Closing of the Sensitive Period in Domestic Chicks (Gallus gallus domesticus)

Filial imprinting of domestic chicks has a well-defined sensitive (critical) period lasting in the laboratory from hatching to day 3. It is a typical model to investigate the molecular mechanisms underlying memory formation in early learning. We recently found that thyroid hormone 3,5,3′-triiodothyronine (T3) is a determinant of the sensitive period. Rapid increases in cerebral T3 levels are induced by imprinting training, rendering chicks imprintable. Furthermore, the administration of exogenous T3 makes chicks imprintable on days 4 or 6 even after the sensitive period has ended. However, how T3 affects neural transmission to enable imprinting remains mostly unknown. In this study, we demonstrate opposing roles for gamma-aminobutyric acid (GABA)-A and GABA-B receptors in imprinting downstream of T3. Quantitative reverse transcription polymerase chain reaction and immunoblotting showed that the GABA-A receptor expression increases gradually from days 1 to 5, whereas the GABA-B receptor expression gradually decreases. We examined whether neurons in the intermediate medial mesopallium (IMM), the brain region responsible for imprinting, express both types of GABA receptors. Immunostaining showed that morphologically identified putative projection neurons express both GABA-A and GABA-B receptors, suggesting that those GABA receptors interact with each other in these cells to modulate the IMM outputs. The roles of GABA-A and GABA-B receptors were investigated using various agonists and antagonists. Our results show that GABA-B receptor antagonists suppressed imprinting on day 1, while its agonists made day 4 chicks imprintable without administration of exogenous T3. By contrast, GABA-A receptor agonists suppressed imprinting on day 1, while its antagonists induced imprintability on day 4 without exogenous T3. Furthermore, both GABA-A receptor agonists and GABA-B receptor antagonists suppressed T3-induced imprintability on day 4 after the sensitive period has ended. Our data from these pharmacological experiments indicate that GABA-B receptors facilitate imprinting downstream of T3 by initiating the sensitive period, while the GABA-A receptor contributes to the termination of the sensitive period. In conclusion, we propose that opposing roles of GABA-A and GABA-B receptors in the brain during development determine the induction and termination of the sensitive period

Endoplasmic Reticulum Stress Contributes to Helicobacter Pylori VacA-Induced Apoptosis

Vacuolating cytotoxin A (VacA) is one of the important virulence factors produced by H. pylori. VacA induces apoptotic cell death, which is potentiated by ammonia. VacA also causes cell death by mitochondrial damage, via signaling pathways that are not fully defined. Our aim was to determine whether endoplasmic reticulum (ER) stress is associated with VacA-induced mitochondrial dysfunction and apoptosis. We found that C/EBP homologous protein (CHOP), a key signaling protein of ER stress-induced apoptosis, was transcriptionally up-regulated following incubation of gastric epithelial cells with VacA. The effect of VacA on CHOP induction was significantly enhanced by co-incubation with ammonium chloride. Phosphorylation of eukaryotic initiation factor 2 (eIF2)-alpha, which is known to occur downstream of the ER stress sensor PKR-like ERlocalized eIF2-alpha kinase (PERK) and to regulate CHOP expression, was also observed following incubation with VacA in the presence of ammonium chloride. Knockdown of CHOP by siRNA resulted in inhibition of VacA-induced apoptosis. Further studies showed that silencing of the PERK gene with siRNA attenuated VacA-mediated phosphorylation of eIF2-alpha, CHOP induction, expression of BH3-only protein Bim and Bax activation, and cell death induced by VacA with ammonium chloride, indicating that ER stress may lead to mitochondrial dysfunction during VacA-induced toxicity. Activation of ER stress and up-regulation of BH3-only proteins were also observed in human H. pylori-infected gastric mucosa. Collectively, this study reveals a possible association between VacA-induced apoptosis in gastric epithelial cells, and activation of ER stress in H. pylori-positive gastric mucosa

Involvement of CD11b+ GR-1low cells in autoimmune disorder in MRL-Faslpr mouse

金沢大学医薬保健研究域医学系Objective: Myeloid-derived suppressor cells (MDSCs) have been identified as immunosuppressive cells in tumor-related inflammation. However, the pathogenesis of MDSCs for autoimmune disease has not been investigated as yet. The aim of this study was to address whether MDSCs contribute to autoimmune organ injury in lupus-prone mice. Methods: MDSCs were analyzed by flow cytometric staining of CD11b+ GR-1+ in MRL-Faslpr mice. CD4+ T-cell proliferation assay was performed by coculture with CD11b+ GR-1+ splenocytes. The percentage of immunosuppressive cells was examined during disease progression. Expression of chemokine receptor on immunosuppressive cells was analyzed, and chemotaxis assay was performed. Results: CD11b+ GR-1low cells had a suppressive effect on CD4+ T-cell proliferation, which was restored by an arginase-1 inhibitor. CD11b+ GR-1low cells increased in percentage during disease progression in kidney and blood. The number of migrated CD11b+ GR-1low cells increased in the presence of monocyte chemoattractant protein-1/CCL2. Conclusion: We assessed the involvement of CD11b+ GR-1low cells in autoimmune disorder in MRL-Faslpr mice. These cells regulate immunological responses via CCL2/CCR2 signaling. The regulation of immunosuppressive monocytes may provide novel therapeutic strategy for organ damage in autoimmune diseases. © 2010 Japanese Society of Nephrology.

Preference for biological motion in domestic chicks: sex-dependent effect of early visual experience

To examine the effects of early visual experience on preference for biological motion (BM), newly hatched chicks were exposed to a point-light animation (a visual stimulus composed of identical light points) depicting the following features of a hen: a walking hen (a BM stimulus), a rotating hen (a non-BM stimulus), a pendulum stimulus, a random motion stimulus and a stationary pattern. Chicks were then tested in a binary choice task, choosing between walking-hen and rotating-hen stimuli. Males exhibited a preference for BM if they had been trained with any animation except the stationary pattern stimulus, suggesting that the BM preference was not learned, but induced by motion stimuli. We found a significant positive correlation between the number of approaches in training and the preference in the test, but locomotion alone did not cause preference for BM. In contrast, females exhibited a particularly strong preference for walking-hen stimuli, but only when they had been trained with it. Furthermore, females (but not males) trained with random motion showed a preference for walking hen over walking cat (a biological motion animation depicting a cat), possibly suggesting that females are choosier than males. Chicks trained with a stationary pattern and untrained controls did not show a significant preference. The induction of BM preference is discussed in terms of possible ecological background of the sex differences

Subjective value of risky foods for individual domestic chicks: a hierarchical Bayesian model

For animals to decide which prey to attack, the gain and delay of the food item must be integrated in a value function. However, the subjective value is not obtained by expected profitability when it is accompanied by risk. To estimate the subjective value, we examined choices in a cross-shaped maze with two colored feeders in domestic chicks. When tested by a reversal in food amount or delay, chicks changed choices similarly in both conditions (experiment 1). We therefore examined risk sensitivity for amount and delay (experiment 2) by supplying one feeder with food of fixed profitability and the alternative feeder with high- or low-profitability food at equal probability. Profitability varied in amount (groups 1 and 2 at high and low variance) or in delay (group 3). To find the equilibrium, the amount (groups 1 and 2) or delay (group 3) of the food in the fixed feeder was adjusted in a total of 18 blocks. The Markov chain Monte Carlo method was applied to a hierarchical Bayesian model to estimate the subjective value. Chicks undervalued the variable feeder in group 1 and were indifferent in group 2 but overvalued the variable feeder in group 3 at a population level. Re-examination without the titration procedure (experiment 3) suggested that the subjective value was not absolute for each option. When the delay was varied, the variable option was often given a paradoxically high value depending on fixed alternative. Therefore, the basic assumption of the uniquely determined value function might be questioned

Biological motion facilitates filial imprinting

To study the functional role of the predisposed preference for Johansson's biological motion (BM) at an early stage of life, newly hatched domestic chicks, Gallus gallus domesticus, were exposed to a variety of motion pictures composed of light points (in red or yellow), and then tested for their learned colour preference. Point-light animations depicting the BM of a walking hen successfully facilitated both the approach activity during imprinting and the learned preference in the test, although significant positive correlations did not appear between these at the individual level. Furthermore, scrambling the light points did not significantly reduce the effects, whereas linear motion of a hen-shaped set of points had no effect. If pretreated with the linear motion, those chicks primed with a high BM preference score showed a high learning score in subsequent imprinting. We conclude that the local movement feature of the BM animation is critical in making chicks approach and learn the associated colour. We propose a scenario wherein naïve chicks have an innate preference for BM, which arises prior to imprinting through nonspecific visual experience in the early posthatch period. The induced BM preference then allows chicks to form a learned colour preference for the associated colour more effectively, leading to the development of tighter social attachment

Competitor suppresses neuronal representation of food reward in the nucleus accumbens/medial striatum of domestic chicks

To investigate the role of social contexts in controlling the neuronal representation of food reward, we recorded single neuron activity in the medial striatum/nucleus accumbens of domestic chicks and examined whether activities differed between two blocks with different contexts. Chicks were trained in an operant task to associate light-emitting diode color cues with three trial types that differed in the type of food reward: no reward (S−), a small reward/short-delay option (SS), and a large reward/long-delay alternative (LL). Amount and duration of reward were set such that both of SS and LL were chosen roughly equally. Neurons showing distinct cue-period activity in rewarding trials (SS and LL) were identified during an isolation block, and activity patterns were compared with those recorded from the same neuron during a subsequent pseudo-competition block in which another chick was allowed to forage in the same area, but was separated by a transparent window. In some neurons, cue-period activity was lower in the pseudo-competition block, and the difference was not ascribed to the number of repeated trials. Comparison at neuronal population level revealed statistically significant suppression in the pseudo-competition block in both SS and LL trials, suggesting that perceived competition generally suppressed the representation of cue-associated food reward. The delay- and reward-period activities, however, did not significantly different between blocks. These results demonstrate that visual perception of a competitive forager per se weakens the neuronal representation of predicted food reward. Possible functional links to impulse control are discussed