pupillometry-to-assess-auditory-sensation-in-guinea-pigs

Pupillometry to Assess Auditory Sensation in Guinea Pigs

Noise exposure is a leading cause of sensorineural hearing loss. Animal models of noise-induced hearing loss have generated mechanistic insight into the ...

University of Pittsburgh

University of Santiago de Compostela

Humans and vocal animals use vocalizations to communicate with members of their species. A necessary function of auditory perception is to generalize across the high variability inherent in vocalization production and classify them into behaviorally distinct categories ('words' or 'call types'). Here, we demonstrate that detecting mid-level features in calls achieves production-invariant classification. Starting from randomly chosen marmoset call features, we use a greedy search algorithm to determine the most informative and least redundant features necessary for call classification. High classification performance is achieved using only 10-20 features per call type. Predictions of tuning properties of putative feature-selective neurons accurately match some observed auditory cortical responses. This feature-based approach also succeeds for call categorization in other species, and for other complex classification tasks such as caller identification. Our results suggest that high-level neural representations of sounds are based on task-dependent features optimized for specific computational goals.

Optimal features for auditory categorization.

Estimates of detection and discrimination thresholds are often used to explore broad perceptual similarities between human subjects and animal models. Pupillometry shows great promise as a non-invasive, easily-deployable method of comparing human and animal thresholds. Using pupillometry, previous studies in animal models have obtained threshold estimates to simple stimuli such as pure tones, but have not explored whether similar pupil responses can be evoked by complex stimuli, what other stimulus contingencies might affect stimulus-evoked pupil responses, and if pupil responses can be modulated by experience or short-term training. In this study, we used an auditory oddball paradigm to estimate detection and discrimination thresholds across a wide range of stimuli in guinea pigs. We demonstrate that pupillometry yields reliable detection and discrimination thresholds across a range of simple (tones) and complex (conspecific vocalizations) stimuli; that pupil responses can be robustly evoked using different stimulus contingencies (low-level acoustic changes, or higher level categorical changes); and that pupil responses are modulated by short-term training. These results lay the foundation for using pupillometry as a reliable method of estimating thresholds in large experimental cohorts, and unveil the full potential of using pupillometry to explore broad similarities between humans and animal models.

Pupillometry as a reliable metric of auditory detection and discrimination across diverse stimulus paradigms in animal models.

Early in auditory processing, neural responses faithfully reflect acoustic input. At higher stages of auditory processing, however, neurons become selective for particular call types, eventually leading to specialized regions of cortex that preferentially process calls at the highest auditory processing stages. We previously proposed that an intermediate step in how nonselective responses are transformed into call-selective responses is the detection of informative call features. But how neural selectivity for informative call features emerges from nonselective inputs, whether feature selectivity gradually emerges over the processing hierarchy, and how stimulus information is represented in nonselective and feature-selective populations remain open question. In this study, using unanesthetized guinea pigs (GPs), a highly vocal and social rodent, as an animal model, we characterized the neural representation of calls in 3 auditory processing stages-the thalamus (ventral medial geniculate body (vMGB)), and thalamorecipient (L4) and superficial layers (L2/3) of primary auditory cortex (A1). We found that neurons in vMGB and A1 L4 did not exhibit call-selective responses and responded throughout the call durations. However, A1 L2/3 neurons showed high call selectivity with about a third of neurons responding to only 1 or 2 call types. These A1 L2/3 neurons only responded to restricted portions of calls suggesting that they were highly selective for call features. Receptive fields of these A1 L2/3 neurons showed complex spectrotemporal structures that could underlie their high call feature selectivity. Information theoretic analysis revealed that in A1 L4, stimulus information was distributed over the population and was spread out over the call durations. In contrast, in A1 L2/3, individual neurons showed brief bursts of high stimulus-specific information and conveyed high levels of information per spike. These data demonstrate that a transformation in the neural representation of calls occurs between A1 L4 and A1 L2/3, leading to the emergence of a feature-based representation of calls in A1 L2/3. Our data thus suggest that observed cortical specializations for call processing emerge in A1 and set the stage for further mechanistic studies.

Neuronal selectivity to complex vocalization features emerges in the superficial layers of primary auditory cortex.

For gaining insight into general principles of auditory processing, it is critical to choose model organisms whose set of natural behaviors encompasses the processes being investigated. This reasoning has led to the development of a variety of animal models for auditory neuroscience research, such as guinea pigs, gerbils, chinchillas, rabbits, and ferrets; but in recent years, the availability of cutting-edge molecular tools and other methodologies in the mouse model have led to waning interest in these unique model species. As laboratories increasingly look to include in-vivo components in their research programs, a comprehensive description of procedures and techniques for applying some of these modern neuroscience tools to a non-mouse small animal model would enable researchers to leverage unique model species that may be best suited for testing their specific hypotheses. In this manuscript, we describe in detail the methods we have developed to apply these tools to the guinea pig animal model to answer questions regarding the neural processing of complex sounds, such as vocalizations. We describe techniques for vocalization acquisition, behavioral testing, recording of auditory brainstem responses and frequency-following responses, intracranial neural signals including local field potential and single unit activity, and the expression of transgenes allowing for optogenetic manipulation of neural activity, all in awake and head-fixed guinea pigs. We demonstrate the rich datasets at the behavioral and electrophysiological levels that can be obtained using these techniques, underscoring the guinea pig as a versatile animal model for studying complex auditory processing. More generally, the methods described here are applicable to a broad range of small mammals, enabling investigators to address specific auditory processing questions in model organisms that are best suited for answering them.

Article	Year
Optimal features for auditory categorization. Nature communications\| PubMed ID: 30899018	2019
Pupillometry as a reliable metric of auditory detection and discrimination across diverse stimulus paradigms in animal models. Scientific reports\| PubMed ID: 33542266	2021
Neuronal selectivity to complex vocalization features emerges in the superficial layers of primary auditory cortex. PLoS biology\| PubMed ID: 34133413	2021
Updates to the guinea pig animal model for in-vivo auditory neuroscience in the low-frequency hearing range. Hearing research\| PubMed ID: 36099806	2022

Pilar Montes-Lourido

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