T Panagiotaropoulos

Cortical Microcircuit Dynamics Mediating Binocular Rivalry: The Role of Adaptation in Inhibition

Perceptual bistability arises when two conflicting interpretations of an ambiguous stimulus or images in binocular rivalry (BR) compete for perceptual dominance. From a computational point of view, competition models based on cross-inhibition and adaptation have shown that noise is a crucial force for rivalry, and operates in balance with adaptation. In particular, noise-driven transitions and adaptation-driven oscillations define two dynamical regimes and the system explains the observed alternations in perception when it operates near their boundary. In order to gain insights into the microcircuit dynamics mediating spontaneous perceptual alternations, we used a reduced recurrent attractor-based biophysically realistic spiking network, well known for working memory, attention, and decision making, where a spike-frequency adaptation mechanism is implemented to account for perceptual bistability. We thus derived a consistently reduced four-variable population rate model using mean-field techniques, and we tested it on BR data collected from human subjects. Our model accounts for experimental data parameters such as mean time dominance, coefficient of variation, and gamma distribution fit. In addition, we show that our model operates near the bifurcation that separates the noise-driven transitions regime from the adaptation-driven oscillations regime, and agrees with Levelt’s second revised and fourth propositions. These results demonstrate for the first time that a consistent reduction of a biophysically realistic spiking network of leaky integrate-and-fire neurons with spike-frequency adaptation could account for BR. Moreover, we demonstrate that BR can be explained only through the dynamics of competing neuronal pools, without taking into account the adaptation of inhibitory interneurons. However, the adaptation of interneurons affects the optimal parametric space of the system by decreasing the overall adaptation necessary for the bifurcation to occur, and introduces oscillations in the spontaneous state.

Introduction to research topic – binocular rivalry: a gateway to studying consciousness

In 1593, Neapolitan polymath Giambattista della Porta publicly lamented that he was unable to improve his impressive productivity (he had published in areas as diverse as cryptography, hydraulics, pharmacology, optics, and classic fiction). Della Porta was trying to read two books simultaneously by placing both volumes side-by-side, and using each eye independently. To his great surprise, his setup allowed him to only read one book at a time. This discovery arguably marks the first written account of binocular rivalry (Wade, 2000) – a perceptual phenomenon that more than 400 years later still both serves to intrigue as well as to illuminate the limits of scientific knowledge. At first glance, binocular rivalry is an oddball. In every day vision, our eyes receive largely matching views of the world. The brain combines the two images into a cohesive scene, and concurrently, perception is stable. However, when showing two very different images (such as two different books) to each eye, the brain resolves the conflict by adopting a “diplomatic” strategy. Rather than mixing the views of the two eyes into an insensible visual percept, observers perceive a dynamically changing series of perceptual snapshots, with one eye’s view dominating for a few seconds before being replaced by its rival from the other eye. With prolonged viewing of a rivalrous stimulus, one inevitably experiences a sequence of subjective perceptual reversals, separated by random time intervals, and this process continues for as long as the sensory conflict is present.

Is the frontal lobe involved in conscious perception

When studying the neural mechanisms underlying conscious perception we should be careful not to misinterpret evidence, and delineate these mechanisms from activity which could reflect the prerequisites or consequences of conscious experiences (Aru et al., 2012; De Graaf et al., 2012). However, at the same time, we need to be careful not to exclude any relevant evidence about the phenomenon. Recently, novel paradigms have attempted to dissociate activity related to conscious perception from activity reflecting its prerequisites and consequences. In particular, one of these studies focused on resolving the role of frontal lobe in conscious perception (Frassle et al., 2014). Through a clever experimental design that contrasted blood-oxygen-level-dependent (BOLD) activity elicited during binocular rivalry with and without behavioral reports, Frassle et al. (2014) suggested that frontal lobe, or a large part of it, may not be necessary for conscious perception per se. Rather frontal areas are involved in processing the consequences of conscious perception like monitoring the perceptual content in order to elicit the appropriate report of the subjective experience. In particular, Frassle et al. showed that behavioral reports of conscious experiences resulted in increased and more widespread activity of the frontal lobe compared to a condition without behavioral reports, where spontaneous transitions in the content of consciousness were estimated through the objective measures like optokinetic nystagmus (OKN) and pupil dilation. The authors of this study concluded that “frontal areas are associated with active report and introspection rather than with rivalry per se.” Therefore, activity in prefrontal regions could be considered as a consequence rather than a direct neural correlate of conscious experience. However, a previous study (Panagiotaropoulos et al., 2012) that measured directly neural activity in the macaque lateral prefrontal cortex (LPFC) using extracellular electrophysiological recordings could help to narrow down the role of frontal activity in conscious perception and exclude the contribution of cognitive or motor consequences in prefrontal neural activity during visual awareness. Specifically, the activity of feature selective neurons in the macaque LFPC was shown to be modulated in accordance with the content of subjective perception, without any confound from motor action (i.e., behavioral reports). Using binocular flash suppression (BFS), a paradigm of robust, externally induced perceptual suppression and without any requirement of behavioral reports, neurons in the LPFC were found to increase or decrease their discharge activity when their preferred stimulus was perceptually dominant or suppressed, respectively. Therefore, since neuronal discharges in the LPFC follow the content of conscious perception even without any motor action, the conclusion of Frassle et al. (2014) about the role of frontal lobe activity in rivalrous perception needs to be refined. Prefrontal activity can indeed reflect the content of conscious perception under conditions of rivalrous stimulation and this activity should not be necessarily considered as the result of a motor action or self-monitoring required for active report. Moreover, the results obtained by Frassle et al. (2014) do not anatomically preclude the entire prefrontal cortex from having a role in conscious perception. Specifically, the BOLD activity related to rivalry in their experiment is still present in the right inferior frontal lobe and right superior frontal lobe (Zaretskaya and Narinyan, 2014). Further, activation of dorso- LPFC in conscious perception of Mooney images was also reported in a study that explicitly controlled for activity elicited by motor action (Imamoglu et al., 2012). It is true that the BFS-related prefrontal activity cannot conclude on a mechanistic, causal involvement of prefrontal activity in driving spontaneous transitions in conscious perception. This is because BFS is a paradigm of externally induced perceptual suppression and is therefore not directly informative about the role of recorded activity in spontaneous transitions. Therefore, the possibility remains open that the kind of prefrontal activity observed in the macaque LPFC during BFS is not a causal factor for conscious perception but rather reflects some other aspects of monitoring that are not directly related to motor action. For example, prefrontal activity could just reflect a read-out from other areas like the inferior temporal cortex (Sheinberg and Logothetis, 1997) that also reliably reflects the content of conscious perception. However, if this is the case, it triggers the question why this activity that closely follows the content of subjective perception is observed in the LPFC even in the absence of any behavioral report. Overall, it motivates further investigation to understand whether prefrontal activity has a mechanistic role in conscious perception or it might underlie some monitoring functions that are not necessarily bound to motor action. Similar to this debate on the role of LPFC in visual awareness, the last decade witnessed disagreement on whether activity in primary visual cortex reflects subjective perception as monitored with electrophysiology and fMRI (Leopold and Logothetis, 1996; Tong, 2003; Maier et al., 2008; Keliris et al., 2010; Leopold, 2012). Measuring both electrophysiological activity and the BOLD signal in the same macaques engaged in an identical task of perceptual suppression finally provided the solution (Maier et al., 2008; Leopold, 2012). Therefore, in order to investigate and resolve the role of PFC in visual perception, one must take a similar approach that utilizes multiple measurement techniques simultaneously or in the same animal along with a careful experimental design. The experimental tasks should not only segregate the effect of various cognitive processes such as attention or introspection in comparison to awareness (Watanabe et al., 2011; Frassle et al., 2014), but also use an objective criterion to decode the content of conscious experience (Frassle et al., 2014), therefore separating perception-related activities from the subsequent behavioral report. Such an approach could therefore robustly delineate the prerequisites and consequences of conscious experience and reveal the true correlates of conscious perception. Lastly, although such a multimodal approach could provide us substantial insights into the activity underlying the representation of conscious content, whether or not this activity has a causal role in mediating perception remains to be understood. Although a number of studies indeed point to a causal involvement of prefrontal cortex in conscious perception (reviewed in Dehaene and Changeux, 2011), a systematic study which directly interferes with prefrontal activity during a task of subjective perception is currently, to the best of our knowledge, missing. While utilizing objective criteria as indicators of perceptual transitions, systematic perturbation of the PFC (such as cooling, transcranial magnetic stimulation, microstimulation, or optogenetics) and observing concomitant changes in the temporal dynamics of perceptual transitions could reveal its causal contribution. Indeed, patients with frontal lesions are impaired in their ability to switch from one subjective view of an ambiguous figure to the other (for example see Ricci and Blundo, 1990, but also see a different case study from Valle-Inclan and Gallego, 2006). We would like to conclude that in formulating our conclusions related to prerequisites, consequences and true correlates of conscious experiences, we need to have an integrative view on the available evidence. Our investigations and conclusions about the neural correlates of consciousness must not only entail better-designed experiments but also diverse experimental techniques (e.g., BOLD fMRI, electrophysiology) that could measure brain activity on different spatial and temporal scales (Panagiotaropoulos et al., 2014). Such a multi-modal approach holds great promise in refining our current understanding of conscious processing.

Visually Driven Activation in Macaque Areas V2 and V3 without Input from the Primary Visual Cortex

Creating focal lesions in primary visual cortex (V1) provides an opportunity to study the role of extra-geniculo-striate pathways for activating extrastriate visual cortex. Previous studies have shown that more than 95% of neurons in macaque area V2 and V3 stop firing after reversibly cooling V1 [1], [2], [3]. However, no studies on long term recovery in areas V2, V3 following permanent V1 lesions have been reported in the macaque. Here we use macaque fMRI to study area V2, V3 activity patterns from 1 to 22 months after lesioning area V1. We find that visually driven BOLD responses persist inside the V1-lesion projection zones (LPZ) of areas V2 and V3, but are reduced in strength by ∼70%, on average, compared to pre-lesion levels. Monitoring the LPZ activity over time starting one month following the V1 lesion did not reveal systematic changes in BOLD signal amplitude. Surprisingly, the retinotopic organization inside the LPZ of areas V2, V3 remained similar to that of the non-lesioned hemisphere, suggesting that LPZ activation in V2, V3 is not the result of input arising from nearby (non-lesioned) V1 cortex. Electrophysiology recordings of multi-unit activity corroborated the BOLD observations: visually driven multi-unit responses could be elicited inside the V2 LPZ, even when the visual stimulus was entirely contained within the scotoma induced by the V1 lesion. Restricting the stimulus to the intact visual hemi-field produced no significant BOLD modulation inside the V2, V3 LPZs. We conclude that the observed activity patterns are largely mediated by parallel, V1-bypassing, subcortical pathways that can activate areas V2 and V3 in the absence of V1 input. Such pathways may contribute to the behavioral phenomenon of blindsight.

Effect of neonatal handling and sex on basal and chronic stress-induced corticosterone and leptin secretion.

Neonatal handling is an experimental paradigm for early experiences. It affects the programming of hypothalamo-pituitary-adrenal (HPA) axis function, known to be sexually dimorphic. Recently leptin, a hormone related to energy balance and secreted mainly by adipocytes, has been implicated in the stress response. We thus determined the effect of neonatal handling on plasma concentrations of corticosterone and leptin of male and female rats under basal conditions and after two consecutive chronic stressors: chronic forced swimming stress and chronic restraint. Handling resulted in lower basal corticosterone levels in both males and females and in a more efficient HPA response, with a large corticosterone surge following the first chronic stressor and a return to basal levels following the second. Handling also resulted in decreased plasma leptin concentrations in males, thus abolishing the sex difference in leptin levels. Furthermore, handling increased body weight while it decreased food intake under basal conditions. Food intake and body weight gain during chronic forced swimming was lower in handled than in non-handled males, while in females these parameters were not influenced by handling. In both males and females, handling resulted in decreased food intake and increased body weight loss during chronic restraint stress. Body weight loss during chronic restraint stress, which is considered an index of maladaptation and ‘depression’, was particularly pronounced in the handled females. Our results also showed that non-handled females had higher corticosterone and lower leptin levels than males under basal conditions and following each of the two chronic stressors. The present work suggests that early experiences, such as the mother–infant relationship, interact with endogenous factors, such as gonadal hormones, to determine the organism’s response to stressful stimuli during adulthood.

Effect of neonatal handling on adult rat spatial learning and memory following acute stress

Brief neonatal handling permanently alters hypothalamic-pituitary-adrenal axis function resulting in increased ability to cope with stress. Since stress is known to affect cognitive abilities, in the present study we investigated the effect of brief (15 min) handling on learning and memory in the Morris water maze, following exposure to an acute restraint stress either before training or recall. Exposure of non-handled rats to the acute stress prior to training resulted in quicker learning of the task, than in the absence of the stressor. When acute stress preceded acquisition, male handled rats showed an overall better learning performance, and both sexes of handled animals were less impaired in the subsequent memory trial, compared to the respective non-handled. In addition, the number of neurons immunoreactive for GR was higher in all areas of Ammons horn of the handled rats during the recall. In contrast, the number of neurons immunoreactive for MR was higher in the CA1 and CA2 areas of the non-handled males. When the acute restraint stress was applied prior to the memory test, neonatal handling was not effective in preventing mnemonic impairment, as all animal groups showed a similar deficit in recall. In this case, no difference between handled and non-handled rats was observed in the number of GR positive neurons in the CA2 and CA3 hippocampal areas during the memory test. These results indicate that early experience interacts with sex and acute stress exposure in adulthood to affect performance in the water maze. Hippocampal corticosterone receptors may play a role in determining the final outcome.

Effects of an Early Experience Involving Training in a T-Maze Under either Denial or Receipt of Expected Reward through Maternal Contact

The mother is the most salient stimulus for the developing pups and a number of early experience models employ manipulation of the mother-infant interaction. We have developed a new model which in addition to changes in maternal behavior includes a learning component on the part of the pups. More specifically, pups were trained in a T-maze and either received (RER rats) or were denied (DER) the reward of maternal contact, during postnatal days 10–13. Pups of both experimental groups learn the T-maze, but the RER do so more efficiently utilizing a procedural-type of learning and memory with activation of the dorsal basal ganglia. On the other hand, the DER experience leads to activation of the hippocampus, prefrontal cortex, and amygdala in the pups. In adulthood, male DER animals exhibit better mnemonic abilities in the Morris water maze and higher activation of the hippocampus, while they have decreased brain serotonergic activity, exhibit a depressive-like phenotype and proactive aggressive behavior in the resident-intruder test. While male RER animals assume a reactive coping style in this test, and showed increased freezing during both contextual and cued memory recall following fear conditioning.

Impairment of learning and memory in TAG-1 deficient mice associated with shorter CNS internodes and disrupted juxtaparanodes.

The cell adhesion molecule TAG-1 is expressed by neurons and glial cells and plays a role in axon outgrowth, migration and fasciculation during development. TAG-1 is also required for the clustering of Kv1.1/1.2 potassium channels and Caspr2 at the juxtaparanodes of myelinated fibers. Behavioral examination of TAG-1 deficient mice (Tag-1(-/-)) showed cognitive impairments in the Morris water maze and novel object recognition tests, reduced spontaneous motor activity, abnormal gait coordination and increased response latency to noxious stimulation. Investigation at the molecular level revealed impaired juxtaparanodal clustering of Caspr2 and Kv1.1/1.2 in the hippocampus, entorhinal cortex, cerebellum and olfactory bulb, with diffusion into the internode. Caspr2 and Kv1.1 levels were reduced in the cerebellum and olfactory bulb. Moreover, Tag-1(-/-) mice had shorter internodes in the cerebral and cerebellar white matter. The detected molecular alterations may account for the behavioural deficits and hyperexcitability in these animals.

Neonatal handling and gender modulate brain monoamines and plasma corticosterone levels following repeated stressors in adulthood

Neonatal handling affects the response to repeated stress in a sexually dimorphic manner. In order to elucidate the mechanisms underlying these gender-dependent effects, we investigated the consequences of neonatal androgenization and handling on adult stress reactivity by determining: (a) immobility time during repeated forced swimming, (b) plasma corticosterone levels, and (c) brain serotonin and dopamine levels and turnover after either repeated forced swimming, or repeated forced swimming followed by repeated restraint stress. In neonatally androgenized females, immobility time was lower in the handled than in the non-handled rats, a pattern resembling that of the males, suggesting that the sexually dimorphic effect of handling on immobility time can be attributed to the organizational effects of testosterone. No differences were found between androgenized females and females injected neonatally with vehicle, indicating that the gender differences in circulating corticosterone are not due to the organizational effects of testosterone. The stress of a neonatal injection interacted with neonatal handling resulting in lower plasma corticosterone and hypothalamic dopamine and serotonin levels in the neonatally injected handled animals following repeated forced swimming. The serotonergic system appears to be sensitive to both the organizational actions of testosterone and the effects of handling, since handled androgenized females had higher serotonin levels and decreased turnover following repeated forced swimming stress, compared to those injected neonatally with vehicle. Handling resulted in increased hypothalamic and striatal serotonin levels in both males and females following repeated forced swimming. Our results reveal that handling has gender-dependent effects on adult hypothalamic-pituitary-adrenal axis and brain monoaminergic system reactivity to stress and that these effects can be attributed to both the organizational and activational effects of gonadal hormones.

Learning of a T-maze by rat pups when contact with the mother is either permitted or denied.

Mother-pup interactions constitute an important component of environmental stimulation of the offspring during the neonatal period. Employing maternal contact as either a positive reinforcer or, its denial, as a frustrative, non-rewarding stimulus, we developed a novel experimental paradigm involving learning by rat neonates of a T-maze. When trained under the reward of maternal contact during postnatal days 10-13 Wistar rat pups learned the choice leading to the mother in a T-maze. When tested 2h later, in the absence of the mother, pups showed a clear preference for the arm of the T-maze leading to the position of the mother during training. Furthermore, pups receiving the expected reward of maternal contact had higher numbers of c-Fos immunopositive cells in the dorsal striatum compared to either naïve or pups denied the expected reward. The above behavioral and cellular results indicate that pups receiving the expected reward developed a procedural-like memory. When trained under frustrative non-reward pups learned to make the correct choice in the T-maze, albeit less efficiently than pups receiving the expected reward. Following this training condition c-Fos immunohistochemistry revealed increased activation of the CA1 area of the hippocampus and the orbitofrontal cortex. Expression of the information learned by the pups denied the expected reward was contingent upon the presence of the mother in the experimental setup in exactly the same configuration as during the training.