electrophysiology-of-laminar-cortical-activity-in-the-common-marmoset

Electrophysiology of Laminar Cortical Activity in the Common Marmoset

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with ...

University of Rochester

The visual pathways that guide actions do not necessarily mediate conscious perception. Patients with primary visual cortex (V1) damage lose conscious perception but often retain unconscious abilities (e.g. blindsight). Here, we asked if saccade accuracy and post-saccadic following responses (PFRs) that automatically track target motion upon saccade landing are retained when conscious perception is lost. We contrasted these behaviors in the blind and intact fields of 11 chronic V1-stroke patients, and in 8 visually intact controls. Saccade accuracy was relatively normal in all cases. Stroke patients also had normal PFR in their intact fields, but no PFR in their blind fields. Thus, V1 damage did not spare the unconscious visual processing necessary for automatic, post-saccadic smooth eye movements. Importantly, visual training that recovered motion perception in the blind field did not restore the PFR, suggesting a clear dissociation between pathways mediating perceptual restoration and automatic actions in the V1-damaged visual system.

Perceptual restoration fails to recover unconscious processing for smooth eye movements after occipital stroke.

The common marmoset (Callithrix jacchus) has been valuable as a primate model in biomedical research. Interest in this species has grown recently, in part due to the successful demonstration of transgenic marmosets. Here we examine the prospects of the marmoset model for visual neuroscience research, adopting a comparative framework to place the marmoset within a broader evolutionary context. The marmoset's small brain bears most of the organizational features of other primates, and its smooth surface offers practical advantages over the macaque for areal mapping, laminar electrode penetration, and two-photon and optical imaging. Behaviorally, marmosets are more limited at performing regimented psychophysical tasks, but do readily accept the head restraint that is necessary for accurate eye tracking and neurophysiology, and can perform simple discriminations. Their natural gaze behavior closely resembles that of other primates, with a tendency to focus on objects of social interest including faces. Their immaturity at birth and routine twinning also makes them ideal for the study of postnatal visual development. These experimental factors, together with the theoretical advantages inherent in comparing anatomy, physiology, and behavior across related species, make the marmoset an excellent model for visual neuroscience.

The marmoset monkey as a model for visual neuroscience.

Smooth pursuit eye movements stabilize slow-moving objects on the retina by matching eye velocity with target velocity. Two critical components are required to generate smooth pursuit: first, because it is a voluntary eye movement, the subject must select a target to pursue to engage the tracking system; and second, generating smooth pursuit requires a moving stimulus. We examined whether this behavior also exists in the common marmoset, a New World primate that is increasingly attracting attention as a genetic model for mental disease and systems neuroscience. We measured smooth pursuit in two marmosets, previously trained to perform fixation tasks, using the standard Rashbass step-ramp pursuit paradigm. We first measured the aspects of visual motion that drive pursuit eye movements. Smooth eye movements were in the same direction as target motion, indicating that pursuit was driven by target movement rather than by displacement. Both the open-loop acceleration and closed-loop eye velocity exhibited a linear relationship with target velocity for slow-moving targets, but this relationship declined for higher speeds. We next examined whether marmoset pursuit eye movements depend on an active engagement of the pursuit system by measuring smooth eye movements evoked by small perturbations of motion from fixation or during pursuit. Pursuit eye movements were much larger during pursuit than from fixation, indicating that pursuit is actively gated. Several practical advantages of the marmoset brain, including the accessibility of the middle temporal (MT) area and frontal eye fields at the cortical surface, merit its utilization for studying pursuit movements.

Motion dependence of smooth pursuit eye movements in the marmoset.

Conversational turn-taking is an integral part of language development, as it reflects a confluence of social factors that mitigate communication. Humans coordinate the timing of speech based on the behaviour of another speaker, a behaviour that is learned during infancy. While adults in several primate species engage in vocal turn-taking, the degree to which similar learning processes underlie its development in these non-human species or are unique to language is not clear. We recorded the natural vocal interactions of common marmosets (Callithrix jacchus) occurring with both their sibling twins and parents over the first year of life and observed at least two parallels with language development. First, marmoset turn-taking is a learned vocal behaviour. Second, marmoset parents potentially played a direct role in guiding the development of turn-taking by providing feedback to their offspring when errors occurred during vocal interactions similarly to what has been observed in humans. Though species-differences are also evident, these findings suggest that similar learning mechanisms may be implemented in the ontogeny of vocal turn-taking across our Order, a finding that has important implications for our understanding of language evolution.

Vocal turn-taking in a non-human primate is learned during ontogeny.

One of the great strengths of the mouse model is the wide array of genetic tools that have been developed. Striking examples include methods for directed modification of the genome, and for regulated expression or inactivation of genes. Within neuroscience, it is now routine to express reporter genes, neuronal activity indicators, and opsins in specific neuronal types in the mouse. However, there are considerable anatomical, physiological, cognitive, and behavioral differences between the mouse and the human that, in some areas of inquiry, limit the degree to which insights derived from the mouse can be applied to understanding human neurobiology. Several recent advances have now brought into reach the goal of applying these tools to understanding the primate brain. Here we describe these advances, consider their potential to advance our understanding of the human brain and brain disorders, discuss bioethical considerations, and describe what will be needed to move forward.

Brains, genes, and primates.

The common marmoset (Callithrix jacchus) has garnered interest recently as a powerful model for the future of neuroscience research. Much of this excitement has centered on the species' reproductive biology and compatibility with gene editing techniques, which together have provided a path for transgenic marmosets to contribute to the study of disease as well as basic brain mechanisms. In step with technical advances is the need to establish experimental paradigms that optimally tap into the marmosets' behavioral and cognitive capacities. While conditioned task performance of a marmoset can compare unfavorably with rhesus monkey performance on conventional testing paradigms, marmosets' social behavior and cognition are more similar to that of humans. For example, marmosets are among only a handful of primates that, like humans, routinely pair bond and care cooperatively for their young. They are also notably pro-social and exhibit social cognitive abilities, such as imitation, that are rare outside of the Apes. In this Primer, we describe key facets of marmoset natural social behavior and demonstrate that emerging behavioral paradigms are well suited to isolate components of marmoset cognition that are highly relevant to humans. These approaches generally embrace natural behavior, which has been rare in conventional primate testing, and thus allow for a new consideration of neural mechanisms underlying primate social cognition and signaling. We anticipate that through parallel technical and paradigmatic advances, marmosets will become an essential model of human social behavior, including its dysfunction in neuropsychiatric disorders.

Marmosets: A Neuroscientific Model of Human Social Behavior.

A 5-y-old multiparous female common marmoset (Callithrix jacchus) presented with acute weight loss of approximately 25% over a 1-wk period. An abdominal mass was apparent on physical examination, and radiographs suggested peritoneal effusion. Exploratory laparotomy revealed hemoperitoneum and an enlarged, gray, hemorrhaging uterus; ovariohysterectomy was performed, and the marmoset recovered. Histologic evaluation of the ovaries and uterus revealed uterine rupture, with invasion of placental villi lined by trophoblasts through the myometrium to the serosal layer. Primary uterine rupture is a rare but serious obstetric event in humans and has been reported only rarely in NHP. This report is the first description of primary uterine rupture during early pregnancy in a common marmoset.

Uterine Rupture in a Common Marmoset (Callithrix jacchus).

Optogenetics has revolutionized the study of functional neuronal circuitry (Boyden ES, Zhang F, Bamberg E, Nagel G, Deisseroth K. Nat Neurosci 8: 1263-1268, 2005; Deisseroth K. Nat Methods 8: 26-29, 2011). Although these techniques have been most successfully implemented in rodent models, they have the potential to be similarly impactful in studies of nonhuman primate brains. Common marmosets (Callithrix jacchus) have recently emerged as a candidate primate model for gene editing, providing a potentially powerful model for studies of neural circuitry and disease in primates. The application of viral transduction methods in marmosets for identifying and manipulating neuronal circuitry is a crucial step in developing this species for neuroscience research. In the present study we developed a novel, chronic method to successfully induce rapid photostimulation in individual cortical neurons transduced by adeno-associated virus to express channelrhodopsin (ChR2) in awake marmosets. We found that large proportions of neurons could be effectively photoactivated following viral transduction and that this procedure could be repeated for several months. These data suggest that techniques for viral transduction and optical manipulation of neuronal populations are suitable for marmosets and can be combined with existing behavioral preparations in the species to elucidate the functional neural circuitry underlying perceptual and cognitive processes.

Optogenetic manipulation of neural circuits in awake marmosets.

The common marmoset has attracted increasing interest as a model for visual neuroscience. A measurement of fundamental importance to ensure the validity of visual studies is spatial acuity. The marmoset has excellent acuity that has been reported at the fovea to be nearly half that of the human (Ordy and Samorajski []: Vision Res 8:1205-1225), a value that is consistent with them having similar photoreceptor densities combined with their smaller eye size (Troilo et al. []: Vision Res 33:1301-1310). Of interest, the marmoset exhibits a higher proportion of cones than rods in peripheral vision than human or macaque, which in principle could endow them with better peripheral acuity depending on how those signals are pooled in subsequent processing. Here, we introduce a simple behavioral paradigm to measure acuity and then test how acuity in the marmoset scales with eccentricity. We trained subjects to fixate a central point and detect a peripheral Gabor by making a saccade to its location. First, we found that accurate assessment of acuity required correction for myopia in all adult subjects. This is an important point because marmosets raised in laboratory conditions often have mild to severe myopia (Graham and Judge []: Vision Res 39:177-187), a finding that we confirm, and that would limit their utility for studies of vision if uncorrected. With corrected vision, we found that their acuity scales with eccentricity similar to that of humans and macaques, having roughly half the value of the human and with no clear departure for higher acuity in the periphery. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 300-313, 2017.

Psychophysical measurement of marmoset acuity and myopia.

Saccadic eye movements sample the visual world and ensure high acuity across the visual field. To compensate for delays in processing, saccades to moving targets require predictions: The eyes must intercept the target's future position to then pursue its direction of motion. Although prediction is crucial to voluntary pursuit, it is unclear whether it is an obligatory feature of saccade planning. Saccade planning involves an involuntary enhanced processing of the target, called presaccadic attention. Does this presaccadic attention recruit smooth eye movements automatically? To test this, we had human participants perform a saccade to one of four apertures, which were static, but each contained a random dot field with motion tangential to the required saccade. In this task, saccades were deviated along the direction of target motion, and the eyes exhibited a following response upon saccade landing. This postsaccadic following response (PFR) increased with spatial uncertainty of the target position and persisted even when we removed the motion stimulus in midflight of the saccade, confirming that it relied on presaccadic information. Motion from 50-100 ms prior to the saccade had the strongest influence on PFR, consistent with the time course of perceptual enhancements reported in presaccadic attention. Finally, the PFR magnitude related linearly to the logarithm of stimulus velocity and generally had low gain, similar to involuntary ocular following movements commonly observed after sudden motion onsets. These results suggest that presaccadic attention selects motion features of targets predictively, presumably to ensure successful immediate tracking of saccade targets in motion.

Presaccadic motion integration drives a predictive postsaccadic following response.

Visual motion processing is a well-established model system for studying neural population codes in primates. The common marmoset, a small new world primate, offers unparalleled opportunities to probe these population codes in key motion processing areas, such as cortical areas MT and MST, because these areas are accessible for imaging and recording at the cortical surface. However, little is currently known about the perceptual abilities of the marmoset. Here, we introduce a paradigm for studying motion perception in the marmoset and compare their psychophysical performance with human observers. We trained two marmosets to perform a motion estimation task in which they provided an analog report of their perceived direction of motion with an eye movement to a ring that surrounded the motion stimulus. Marmosets and humans exhibited similar trade-offs in speed versus accuracy: errors were larger and reaction times were longer as the strength of the motion signal was reduced. Reverse correlation on the temporal fluctuations in motion direction revealed that both species exhibited short integration windows; however, marmosets had substantially less nondecision time than humans. Our results provide the first quantification of motion perception in the marmoset and demonstrate several advantages to using analog estimation tasks.

Motion Perception in the Common Marmoset.

Circadian clocks drive cyclic variations in many aspects of physiology, but some daily variations are evoked by periodic changes in the environment or sleep-wake state and associated behaviors, such as changes in posture, light levels, fasting or eating, rest or activity and social interactions; thus, it is often important to quantify the relative contributions of these factors. Yet, circadian rhythms and these evoked effects cannot be separated under typical 24-h day conditions, because circadian phase and the length of time awake or asleep co-vary. Nathaniel Kleitman's forced desynchrony (FD) protocol was designed to assess endogenous circadian rhythmicity and to separate circadian from evoked components of daily rhythms in multiple parameters. Under FD protocol conditions, light intensity is kept low to minimize its impact on the circadian pacemaker, and participants have sleep-wake state and associated behaviors scheduled to an imposed non-24-h cycle. The period of this imposed cycle, Τ, is chosen so that the circadian pacemaker cannot entrain to it and therefore continues to oscillate at its intrinsic period (τ, ~24.15 h), ensuring circadian components are separated from evoked components of daily rhythms. Here we provide detailed instructions and troubleshooting techniques on how to design, implement and analyze the data from an FD protocol. We provide two procedures: one with general guidance for designing an FD study and another with more precise instructions for replicating one of our previous FD studies. We discuss estimating circadian parameters and quantifying the separate contributions of circadian rhythmicity and the sleep-wake cycle, including statistical analysis procedures and an R package for conducting the non-orthogonal spectral analysis method that enables an accurate estimation of period, amplitude and phase.

Desynchronizing the sleep---wake cycle from circadian timing to assess their separate contributions to physiology and behaviour and to estimate intrinsic circadian period.

Primates have evolved sophisticated, visually guided reaching behaviors for interacting with dynamic objects, such as insects, during foraging. Reaching control in dynamic natural conditions requires active prediction of the target's future position to compensate for visuo-motor processing delays and to enhance online movement adjustments. Past reaching research in non-human primates mainly focused on seated subjects engaged in repeated ballistic arm movements to either stationary targets or targets that instantaneously change position during the movement. However, those approaches impose task constraints that limit the natural dynamics of reaching. A recent field study in marmoset monkeys highlights predictive aspects of visually guided reaching during insect prey capture among wild marmoset monkeys. To examine the complementary dynamics of similar natural behavior within a laboratory context, we developed an ecologically motivated, unrestrained reach-to-grasp task involving live crickets. We used multiple high-speed video cameras to capture the movements of common marmosets (Callithrix jacchus) and crickets stereoscopically and applied machine vision algorithms for marker-free object and hand tracking. Contrary to estimates under traditional constrained reaching paradigms, we find that reaching for dynamic targets can operate at incredibly short visuo-motor delays around 80 ms, rivaling the speeds that are typical of the oculomotor systems during closed-loop visual pursuit. Multivariate linear regression modeling of the kinematic relationships between the hand and cricket velocity revealed that predictions of the expected future location can compensate for visuo-motor delays during fast reaching. These results suggest a critical role of visual prediction facilitating online movement adjustments for dynamic prey.

Fast prediction in marmoset reach-to-grasp movements for dynamic prey.

Fixation constraints in visual tasks are ubiquitous in visual and cognitive neuroscience. Despite its widespread use, fixation requires trained subjects, is limited by the accuracy of fixational eye movements, and ignores the role of eye movements in shaping visual input. To overcome these limitations, we developed a suite of hardware and software tools to study vision during natural behavior in untrained subjects. We measured visual receptive fields and tuning properties from multiple cortical areas of marmoset monkeys who freely viewed full-field noise stimuli. The resulting receptive fields and tuning curves from primary visual cortex (V1) and area MT match reported selectivity from the literature which was measured using conventional approaches. We then combined free viewing with high-resolution eye tracking to make the first detailed 2D spatiotemporal measurements of foveal receptive fields in V1. These findings demonstrate the power of free viewing to characterize neural responses in untrained animals while simultaneously studying the dynamics of natural behavior.

Article	Year
Perceptual restoration fails to recover unconscious processing for smooth eye movements after occipital stroke. eLife\| PubMed ID: 35730931	2022
The marmoset monkey as a model for visual neuroscience. Neuroscience research\| PubMed ID: 25683292	2015
Motion dependence of smooth pursuit eye movements in the marmoset. Journal of neurophysiology\| PubMed ID: 25867740	2015
Vocal turn-taking in a non-human primate is learned during ontogeny. Proceedings. Biological sciences\| PubMed ID: 25904663	2015
Brains, genes, and primates. Neuron\| PubMed ID: 25950631	2015
Marmosets: A Neuroscientific Model of Human Social Behavior. Neuron\| PubMed ID: 27100195	2016
Uterine Rupture in a Common Marmoset (Callithrix jacchus). Comparative medicine\| PubMed ID: 27298252	2016
Optogenetic manipulation of neural circuits in awake marmosets. Journal of neurophysiology\| PubMed ID: 27334951	2016
Psychophysical measurement of marmoset acuity and myopia. Developmental neurobiology\| PubMed ID: 27804251	2017
Presaccadic motion integration drives a predictive postsaccadic following response. Journal of vision\| PubMed ID: 31557762	2019
Motion Perception in the Common Marmoset. Cerebral cortex (New York, N.Y. : 1991)\| PubMed ID: 31828299	2020
Desynchronizing the sleep---wake cycle from circadian timing to assess their separate contributions to physiology and behaviour and to estimate intrinsic circadian period. Nature protocols\| PubMed ID: 36376588	2023
Fast prediction in marmoset reach-to-grasp movements for dynamic prey. Current biology : CB\| PubMed ID: 37279754	2023
Detailed characterization of neural selectivity in free viewing primates. Nature communications\| PubMed ID: 37339973	2023

Jude F. Mitchell

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