Altered pupil light and darkness reflex and eye-blink responses in late-life depression.

BACKGROUND: Late-life depression (LLD) is a prevalent neuropsychiatric disorder in the older population. While LLD exhibits high mortality rates, depressive symptoms in older adults are often masked by physical health conditions. In younger adults, depression is associated with deficits in pupil light reflex and eye blink rate, suggesting the potential use of these responses as biomarkers for LLD. METHODS: We conducted a study using video-based eye-tracking to investigate pupil and blink responses in LLD patients (n = 25), older (OLD) healthy controls (n = 29), and younger (YOUNG) healthy controls (n = 25). The aim was to determine whether there were alterations in pupil and blink responses in LLD compared to both OLD and YOUNG groups. RESULTS: LLD patients displayed significantly higher blink rates and dampened pupil constriction responses compared to OLD and YOUNG controls. While tonic pupil size in YOUNG differed from that of OLD, LLD patients did not exhibit a significant difference compared to OLD and YOUNG controls. GDS-15 scores in older adults correlated with light and darkness reflex response variability and blink rates. PHQ-15 scores showed a correlation with blink rates, while MoCA scores correlated with tonic pupil sizes. CONCLUSIONS: The findings demonstrate that LLD patients display altered pupil and blink behavior compared to OLD and YOUNG controls. These altered responses correlated differently with the severity of depressive, somatic, and cognitive symptoms, indicating their potential as objective biomarkers for LLD.

Linking tonic and phasic pupil responses to P300 amplitude in an emotional face-word Stroop task.

The locus coeruleus-norepinephrine (LC-NE) system, which regulates arousal levels, is important for cognitive control, including emotional conflict resolution. Additionally, the LC-NE system is implicated in P300 generation. If the P300 is mediated by the LC-NE system, and considering the established correlations between LC activity and pupil dilation, P300 amplitude should correlate with task-evoked (phasic) pupil dilation on a trial-by-trial basis. However, prior studies, predominantly utilizing oddball-type paradigms, have not demonstrated correlations between concurrently recorded task-evoked pupil dilation and P300 responses. Using a recently developed emotional face-word Stroop task that links pupil dilation to the LC-NE system, here, we examined both intra- and inter-individual correlations between task-evoked pupil dilation and P300 amplitude. We found that lower accuracy, slower reaction times, and larger task-evoked pupil dilation were obtained in the incongruent compared to the congruent condition. Furthermore, we observed intra-individual correlations between task-evoked pupil dilation and P300 amplitude, with larger pupil dilation correlating with a greater P300 amplitude. In contrast, pupil dilation did not exhibit consistent correlations with N450 and N170 amplitudes. Baseline (tonic) pupil size also showed correlations with P300 and N170 amplitudes, with smaller pupil size corresponding to larger amplitude. Moreover, inter-individual differences in task-evoked pupil dilation between the congruent and incongruent conditions correlated with differences in reaction time and P300 amplitude, though these effects only approached significance. To summarize, our study provides evidence for a connection between task-evoked pupil dilation and P300 amplitude at the single-trial level, suggesting the involvement of the LC-NE system in P300 generation.

Time-on-task effects on human pupillary and saccadic metrics after theta burst transcranial magnetic stimulation over the frontal eye field.

Pupil size undergoes constant changes primarily influenced by ambient luminance. These changes are referred to as the pupillary light reflex (PLR), where the pupil transiently constricts in response to light. PLR kinematics provides valuable insights into autonomic nervous system function and have significant clinical applications. Recent research indicates that attention plays a role in modulating the PLR, and the circuit involving the frontal eye field (FEF) and superior colliculus is causally involved in controlling this pupillary modulation. However, there is limited research exploring the role of the human FEF in these pupillary responses, and its impact on PLR metrics remains unexplored. Additionally, although the protocol of continuous theta-burst stimulation (cTBS) is well-established, the period of disruption after cTBS is yet to be examined in pupillary responses. Our study aimed to investigate the effects of FEF cTBS on pupillary and saccadic metrics in relation to time spent performing a task (referred to as time-on-task). We presented a bright stimulus to induce the PLR in visual- and memory-delay saccade tasks following cTBS over the right FEF or vertex. FEF cTBS, compared to vertex cTBS, resulted in decreased baseline pupil size, peak constriction velocities, and amplitude. Furthermore, the time-on-task effects on baseline pupil size, peak amplitude, and peak time differed between the two stimulation conditions. In contrast, the time-on-task effects on saccadic metrics were less pronounced between the two conditions. In summary, our study provides the first evidence that FEF cTBS affects human PLR metrics and that these effects are modulated by time-on-task.

Human microsaccade cueing modulation in visual- and memory-delay saccade tasks after theta burst transcranial magnetic stimulation over the frontal eye field.

Microsaccades that occur during periods of fixation are modulated by various cognitive processes and have an impact on visual processing. A network of brain areas is involved in microsaccade generation, including the superior colliculus and frontal eye field (FEF) which are involved in modulating microsaccade rate and direction after the appearance of a visual cue (referred to as microsaccade cueing modulation). Although the neural mechanisms underlying microsaccade cueing modulations have been demonstrated in monkeys, limited research has investigated a causal role of these areas in humans. By applying continuous theta-burst transcranial magnetic stimulation (cTBS) over the right FEF and vertex, we investigated the role of human FEF in modulating microsaccade responses after the appearance of a visual target in a visual- and memory-delay saccade task. After target appearance, microsaccade rate was initially suppressed but then increased in both cTBS conditions. More importantly, in the visual-delay task, microsaccades after target appearance were directed to the ipsilateral side more often with FEF, compared to vertex stimulation. Moreover, microsaccades were directed towards the target location, then to the opposite location of the target in both tasks, with larger effects in the visual-, compared to, memory-delay task. This microsaccade direction modulation was delayed after FEF stimulation in the memory-delay task. Overall, some microsaccade cueing modulations were moderately disrupted after FEF cTBS, suggesting a causal role for involvement of the human FEF in microsaccade generation after presentation of salient stimuli.

Investigating the role of human frontal eye field in the pupil light reflex modulation by saccade planning and working memory.

The pupil constricts in response to an increase in global luminance level, commonly referred to as the pupil light reflex. Recent research has shown that these reflex responses are modulated by high-level cognition. There is larger pupil constriction evoked by a bright stimulus when the stimulus location spatially overlaps with the locus of attention, and these effects have been extended to saccade planning and working memory (here referred to as pupil local-luminance modulation). Although research in monkeys has further elucidated a central role of the frontal eye field (FEF) and superior colliculus in the pupil local-luminance modulation, their roles remain to be established in humans. Through applying continuous theta-burst transcranial magnetic stimulation over the right FEF (and vertex) to inhibit its activity, we investigated the role of the FEF in human pupil local-luminance responses. Pupil light reflex responses were transiently evoked by a bright patch stimulus presented during the delay period in the visual- and memory-delay tasks. In the visual-delay task, larger pupil constriction was observed when the patch location was spatially aligned with the target location in both stimulation conditions. More interestingly, after FEF stimulation, larger pupil constriction was obtained when the patch was presented in the contralateral, compared to the ipsilateral visual field of the stimulation. In contrast, FEF stimulation effects were absence in the memory-delay task. Linear mixed model results further found that stimulation condition, patch location consistency, and visual field significantly modulated observed pupil constriction responses. Together, our results constitute the first evidence of FEF modulation in human pupil local-luminance responses.

Fatigue and Arousal Modulations Revealed by Saccade and Pupil Dynamics.

Saccadic eye movements are directed to the objects of interests and enable high-resolution visual images in the exploration of the visual world. There is a trial-to-trial variation in saccade dynamics even in a simple task, possibly attributed to arousal fluctuations. Previous studies have showed that an increase of fatigue level over time, also known as time-on-task, can be revealed by saccade peak velocity. In addition, pupil size, controlled by the autonomic nervous system, has long been used as an arousal index. However, limited research has been done with regards to the relation between pupil size and saccade behavior in the context of trial-to-trial variation. To investigate fatigue and arousal effects on saccadic and pupillary responses, we used bright and emotional stimuli to evoke pupillary responses in tasks requiring reactive and voluntary saccade generation. Decreased voluntary saccade peak velocities, reduced tonic pupil size and phasic pupillary responses were observed as time-on-task increased. Moreover, tonic pupil size affected saccade latency and dynamics, with steeper saccade main sequence slope as tonic pupil size increased. In summary, saccade dynamics and tonic pupil size were sensitive to fatigue and arousal level, together providing valuable information for the understanding of human behavior.

Pupillometry as an integrated readout of distinct attentional networks.

The course of pupillary constriction and dilation provides an easy-to-access, inexpensive, and noninvasive readout of brain activity. We propose a new taxonomy of factors affecting the pupil and link these to associated neural underpinnings in an ascending hierarchy. In addition to two well-established low-level factors (light level and focal distance), we suggest two further intermediate-level factors, alerting and orienting, and a higher-level factor, executive functioning. Alerting, orienting, and executive functioning - including their respective underlying neural circuitries - overlap with the three principal attentional networks, making pupil size an integrated readout of distinct states of attention. As a now widespread technique, pupillometry is ready to provide meaningful applications and constitutes a viable part of the psychophysiological toolbox.

Pupil-linked Arousal Signals in the Midbrain Superior Colliculus.

The orienting response evoked by the appearance of a salient stimulus is modulated by arousal; however, neural underpinnings for the interplay between orienting and arousal are not well understood. The superior colliculus (SC), causally involved in multiple components of the orienting response including gaze and attention shifts, receives not only multisensory and cognitive inputs but also arousal-regulated inputs from various cortical and subcortical structures. To investigate the impact of moment-by-moment fluctuations in arousal on orienting saccade responses, we used microstimulation of the monkey SC to trigger saccade responses, and we used pupil size and velocity to index the level of arousal at stimulation onset because these measures correlate with changes in brain states and locus coeruleus activity. Saccades induced by SC microstimulation correlated with prestimulation pupil velocity, with higher pupil velocities on trials without evoked saccades than with evoked saccades. In contrast, prestimulation absolute pupil size did not correlate with saccade behavior. Moreover, pupil velocity correlated with evoked saccade latency and metrics. Together, our results demonstrated that small fluctuations in arousal, indexed by pupil velocity, can modulate the saccade response evoked by SC microstimulation in awake behaving monkeys.

Pupillary responses to differences in luminance, color and set size.

The pupil responds to a salient stimulus appearing in the environment, in addition to its modulation by global luminance. These pupillary responses can be evoked by visual or auditory stimuli, scaled with stimulus salience, and enhanced by multisensory presentation. In addition, pupil size is modulated by various visual stimulus attributes, such as color, area, and motion. However, research that concurrently examines the influence of different factors on pupillary responses is limited. To explore how presentation of multiple visual stimuli influences human pupillary responses, we presented arrays of visual stimuli and systematically varied their luminance, color, and set size. Saliency level, computed by the saliency model, systematically changed with set size across all conditions, with higher saliency levels in larger set sizes. Pupillary constriction responses were evoked by the appearance of visual stimuli, with larger pupillary responses observed in larger set size. These effects were pronounced even though the global luminance level was unchanged using isoluminant chromatic stimuli. Furthermore, larger pupillary constriction responses were obtained in the blue, compared to other color conditions. Together, we argue that both cortical and subcortical areas contribute to the observed pupillary constriction modulated by set size and color.

Time-frequency analysis of pupil size modulated by global luminance, arousal, and saccade preparation signals using Hilbert-Huang transform.

Pupil size changes constantly and is mainly determined by global luminance signals. In addition, the pupil responds to various cognitive and arousal processes, with larger pupil dilation observed in higher levels of cognitive or arousal processing. Although these task-evoked pupillary responses are extensively used in the pupil research, pupil analysis focusing on the frequency domain, particularly in the context of arousal and cognitive modulations, is less established. Fourier Transform method (FFT) has been used to understand the modulation of task difficulty on pupil oscillations. However, physiological signals are often characterized as non-linear and non-stationary waves, and the conventional spectral analytical method with linearity presumption is less appropriate to reveal modulation dynamics between time and frequency. Here, we used Hilbert-Huang Transform (HHT) to examine the time-frequency modulations on pupil size regulated by arousal, cognitive, and global luminance signals. Consistent with previous research, using FFT, higher spectral densities were obtained with lower luminance background. Moreover, higher spectral densities were found in the high emotional arousal condition. With HHT, we further demonstrated temporal changes on amplitude spectrum and inter-trial phase coherence (ITPC) in each intrinsic mode function (IMF), with stronger amplitudes in higher IMFs (i.e., low frequencies). Moreover, although global luminance, arousal and saccade preparation modulated pupil oscillatory responses, the modulation pattern in different IMFs was different. Together, our results demonstrated dynamics between the time and frequency domain on pupil oscillatory responses, highlighting the importance of examining the time-frequency interactions in the context of various pupil modulations.

Role of the frontal eye field in human microsaccade responses: A TMS study.

Microsaccade is a type of fixational eye movements that is modulated by various sensory and cognitive processes, and impact our visual perception. Although studies in monkeys have demonstrated a functional role for the superior colliculus and frontal eye field (FEF) in controlling microsaccades, our understanding of the neural mechanisms underlying the generation of microsaccades is still limited. By applying continuous theta-burst stimulation (cTBS) over the right FEF and the vertex, we investigated the role of the FEF in generating human microsaccade responses evoked by salient stimuli or by changes in background luminance. We observed higher microsaccade rates prior to target appearance, and larger rebound in microsaccade occurrence following salient stimuli, when disruptive cTBS was applied over FEF compared to vertex stimulation. Moreover, the microsaccade direction modulation after changes in background luminance was disrupted with FEF stimulation. Together, our results constitute the first evidence of FEF modulation in human microsaccade responses.

Differentiating global luminance, arousal and cognitive signals on pupil size and microsaccades.

Pupil size reflects a proxy for neural activity associated with global luminance, arousal and cognitive processing. Microsaccades are also modulated by arousal and cognitive processing. Are these effects of arousal and cognitive signals on pupil size and microsaccades coordinated? If so, via what neural mechanisms? We hypothesized that if pupil size and microsaccades are coordinately modulated by these processes, pupil size immediately before microsaccade onset, as an index for ongoing cognitive and arousal processing, should correlate with microsaccade responses during tasks alternating these signals. Here, we examined the relationship between pupil size and microsaccade responses in tasks that included variations in global luminance, arousal and inhibitory control. Higher microsaccade peak velocities correlated with larger pre-microsaccade pupil response related to arousal and inhibitory control signals. In contrast, pupil responses evoked by global luminance signals did not correlate with microsaccade responses. Given the central role of the superior colliculus in microsaccade generation, these results suggest the critical involvement of the superior colliculus to coordinate pupil and microsaccade responses for arousal and inhibitory control modulations, but not for the pupil luminance modulation.

Linking Pupil Size Modulated by Global Luminance and Motor Preparation to Saccade Behavior.

Saccades are rapid eye movements that are used to move the high acuity fovea in a serial manner in the exploration of the visual scene. Stimulus contrast is known to modulate saccade latency and metrics possibly via changing visual activity in the superior colliculus (SC), a midbrain structure causally involved in saccade generation. However, the quality of visual signals should also be modulated by the amount of lights projected onto the retina, which is gated by the size of the pupil. Although absolute pupil size should modulate visual signals and in turn affect saccade responses, research examining this relationship is very limited. Besides, pupil size is associated with motor preparation. However, the role of pupil dilation in saccade metrics remains unexplored. Through varying peripheral background luminance level and target visual contrast in the saccade task, we investigated the role of absolute pupil size and baseline-corrected pupil dilation in saccade latency and metrics. Higher target detection accuracy was obtained with lower background luminance level, and larger absolute pupil diameter correlated with smaller saccade amplitude and higher saccade peak velocities. More interestingly, the comparable modulation between pupil dilation and stimulus contrast was obtained, showing larger pupil dilation (or higher contrast stimuli) correlating with faster saccade latencies, larger amplitude, higher peak velocities, and smaller endpoint deviation. Together, our results demonstrated the influence of absolute pupil size induced by global luminance level and baseline-corrected pupil dilation associated with motor preparation on saccade latency and metrics, implicating the role of the SC in this behavior.

Coordination of Pupil and Saccade Responses by the Superior Colliculus.

The appearance of a salient stimulus evokes saccadic eye movements and pupil dilation as part of the orienting response. Although the role of the superior colliculus (SC) in saccade and pupil dilation has been established separately, whether and how these responses are coordinated remains unknown. The SC also receives global luminance signals from the retina, but whether global luminance modulates saccade and pupil responses coordinated by the SC remains unknown. Here, we used microstimulation to causally determine how the SC coordinates saccade and pupil responses and whether global luminance modulates these responses by varying stimulation frequency and global luminance in male monkeys. Stimulation frequency modulated saccade and pupil responses, with trial-by-trial correlations between the two responses. Global luminance only modulated pupil, but not saccade, responses. Our results demonstrate an integrated role of the SC on coordinating saccade and pupil responses, characterizing luminance independent modulation in the SC, together elucidating the differentiated pathways underlying this behavior.

Role of the frontal eye field in human pupil and saccade orienting responses.

The appearance of a salient stimulus evokes a series of orienting responses including saccades and pupil size to prepare the body for appropriate action. The midbrain superior colliculus (SC) that receives critical control signals from the frontal eye field (FEF) is hypothesized to coordinate all components of orienting. It has shown recently that the FEF, together with the SC, is also importantly involved in the control of pupil size, in addition to its well-documented role in eye movements. Although the role of the FEF in pupil size is demonstrated in monkeys, its role in human pupil responses and the coordination between pupil size and saccades remains to be established. Through applying continuous theta-burst stimulation over the right FEF and vertex, we investigated the role of the FEF in human pupil and saccade responses evoked by a salient stimulus, and the coordination between pupil size and saccades. Our results showed that neither saccade reaction times (SRT) nor pupil responses evoked by salient stimuli were modulated by FEF stimulation. In contrast, the correlation between pupil size and SRTs in the contralateral stimulus condition was diminished with FEF stimulation, but intact with vertex stimulation. Moreover, FEF stimulation effects between saccade and pupil responses associated with salient stimuli correlated across participants. This is the first transcranial magnetic stimulation (TMS) study on the pupil orienting response, and our findings suggest that human FEF was involved in coordinating pupil size and saccades, but not involved in the control of pupil orienting responses.

Investigating Arousal, Saccade Preparation, and Global Luminance Effects on Microsaccade Behavior.

Microsaccades, small saccadic eye movements occurring during fixation, have been suggested to be modulated by various sensory, cognitive, and affective processes relating to arousal. Although the modulation of fatigue-related arousal on microsaccade behavior has previously been characterized, the influence of other aspects of arousal, such as emotional arousal, is less understood. Moreover, microsaccades are modulated by cognitive processes (e.g., voluntary saccade preparation) that could also be linked to arousal. To investigate the influence of emotional arousal, saccade preparation, and global luminance levels on microsaccade behavior, emotional auditory stimuli were presented prior to the onset of a fixation cue whose color indicated to look either at the peripheral stimulus (pro-saccade) or in the opposite direction of the stimulus (anti-saccade). Microsaccade behavior was found to be significantly modulated by saccade preparation and global luminance level, but not emotional arousal. In the pro- and anti-saccade task, microsaccade rate was lower during anti-saccade preparation as compared to pro-saccade preparation, though microsaccade dynamics were comparable during both trial types. Our results reveal a differential role of arousal linked to emotion, fatigue, saccade preparation, and global luminance level on microsaccade behavior.

Effects of pupillary light and darkness reflex on the generation of pro- And anti-saccades.

Saccades are often directed toward a stimulus that provides useful information for observers to navigate the visual world. The quality of visual signals of a stimulus is influenced by global luminance, and the pupil constricts or dilates after a luminance increase or decrease, respectively, to optimize visual signals for further information processing. Although luminance level changes regularly in the real environment, saccades are mostly studied in the luminance-unchanged setup. Whether pupillary responses triggered by global luminance changes modulate saccadic behavior are yet to be explored. Through varying background luminance level in an interleaved pro- and anti-saccade paradigm, we investigated the modulation of pupillary luminance responses on the generation of reflexive and voluntary saccades. Subjects were instructed to either automatically look at the peripheral stimulus (pro-saccade) or to suppress the automatic response and voluntarily look in the opposite direction from the stimulus (anti-saccade). Level of background luminance was increased (light), decreased (dark), or unchanged (control) during the instructed fixation period. Saccade reaction time distributions of correct pro- and anti-saccades in the light and dark conditions were differed significantly from those in the control condition. Moreover, the luminance condition modulated saccade kinematics, showing reduced performances in the light condition than in the control condition, particularly in pro-saccades. Modeling results further suggested that both pupil diameter and pupil size derivative significantly modulated saccade behavior, though effect sizes were small and mainly mediated by intersubject differences. Together, our results demonstrated the influence of pupillary luminance responses on the generation of pro- and anti-saccades.

Background luminance effects on pupil size associated with emotion and saccade preparation.

Pupil dilation is consistently evoked by affective and cognitive processing, and this dilation can result from sympathetic activation or parasympathetic inhibition. The relative contributions of the sympathetic and parasympathetic systems on the pupillary response induced by emotion and cognition may be different. Sympathetic and parasympathetic activity is regulated by global luminance level. Higher luminance levels lead to greater activation of the parasympathetic system while lower luminance levels lead to greater activation of the sympathetic system. To understand the contributions of the sympathetic and parasympathetic nervous systems to pupillary responses associated with emotion and saccade preparation, emotional auditory stimuli were presented following the fixation cue whose color indicated instruction to perform a pro- or anti-saccade while varying the background luminance level. Pupil dilation was evoked by emotional auditory stimuli and modulated by arousal level. More importantly, greater pupil dilation was observed with a dark background, compared to a bright background. In contrast, pupil dilation responses associated with saccade preparation were larger with the bright background than the dark background. Together, these results suggest that arousal-induced pupil dilation was mainly mediated by sympathetic activation, but pupil dilation related to saccade preparation was primarily mediated by parasympathetic inhibition.

Saliency and priority modulation in a pop-out paradigm: Pupil size and microsaccades.

A salient stimulus can trigger a coordinated orienting response consisting of a saccade, pupil, and microsaccadic responses. Saliency models predict that the degree of visual conspicuity of all visual stimuli guides visual orienting. By presenting a multiple-item array that included an oddball colored item (pop-out), randomly mixed colored items (mixed-color), or single-color items (single-color), we examined the effects of saliency and priority (saliency + relevancy) on pupil size and microsaccade responses. Larger pupil responses were produced in the pop-out compared to the mixed-color or single-color conditions after stimulus presentation. However, the saliency modulation on microsaccades was not significant. Furthermore, although goal-relevancy information did not modulate pupil responses and microsaccade rate, microsaccade direction was biased toward the pop-out item when it was the subsequent saccadic target. Together, our results demonstrate saliency modulation on pupil size and priority effects on microsaccade direction during visual pop-out.

Investigating cognitive load modulation of distractor processing using pupillary luminance responses in the anti-saccade paradigm.

Observers must select goal-directed stimuli in lieu of distractors in the environment for preferential information processing. This selection, according to the load theory of attention, is modulated by cognitive load, involving the frontal cortices, with more significant distractor interference under high cognitive load, with strained executive control resources. Evidence in support of this theory exists; however, working memory tasks were predominately used in these investigations. The influence of other types of cognitive load on distractor processing is largely unknown. An interleaved pro- and anti-saccade task has often been used to investigate executive control in which subjects are instructed in advance to either automatically look at the peripheral stimulus (pro-saccade), or to suppress the automatic response and voluntarily look in the direction opposite of the stimulus (anti-saccade). Distinct frontal preparatory activity has been clearly characterized during preparation for pro- and anti-saccades, with higher inhibition-related activity in preparation for anti-saccades than pro-saccades. Here, we used an interleaved pro- and anti-saccade paradigm to investigate the modulation of distractor interference by cognitive load in a group of 24 healthy young adults. Luminant distractors were used to evoke automatic pupillary responses to evaluate distractor processing. Greater pupillary dilation following dark distractor presentation was observed in the anti-saccade than the pro-saccade preparation. These effects, however, were absent in pupillary constriction following bright distractors. Together, our results support the load theory of attention, importantly highlighting the potential of using involuntary changes in pupil size to objectively investigate attentional selection under load.