Knockdown of the Non-canonical Wnt Gene Prickle2 Leads to Cerebellar Purkinje Cell Abnormalities While Cerebellar-Mediated Behaviors Remain Intact.

Autism spectrum disorders (ASD) involve brain wide abnormalities that contribute to a constellation of symptoms including behavioral inflexibility, cognitive dysfunction, learning impairments, altered social interactions, and perceptive time difficulties. Although a single genetic variation does not cause ASD, genetic variations such as one involving a non-canonical Wnt signaling gene, Prickle2, has been found in individuals with ASD. Previous work looking into phenotypes of Prickle2 knock-out (Prickle2-/-) and heterozygous mice (Prickle2-/+) suggest patterns of behavior similar to individuals with ASD including altered social interaction and behavioral inflexibility. Growing evidence implicates the cerebellum in ASD. As Prickle2 is expressed in the cerebellum, this animal model presents a unique opportunity to investigate the cerebellar contribution to autism-like phenotypes. Here, we explore cerebellar structural and physiological abnormalities in animals with Prickle2 knockdown using immunohistochemistry, whole-cell patch clamp electrophysiology, and several cerebellar-associated motor and timing tasks, including interval timing and eyeblink conditioning. Histologically, Prickle2-/- mice have significantly more empty spaces or gaps between Purkinje cells in the posterior lobules and a decreased propensity for Purkinje cells to fire action potentials. These structural cerebellar abnormalities did not impair cerebellar-associated behaviors as eyeblink conditioning and interval timing remained intact. Therefore, although Prickle-/- mice show classic phenotypes of ASD, they do not recapitulate the involvement of the adult cerebellum and may not represent the pathophysiological heterogeneity of the disorder.

Cerebellar D1DR-expressing neurons modulate the frontal cortex during timing tasks.

Converging lines of evidence suggest that the cerebellum plays an integral role in cognitive function through its interactions with association cortices like the medial frontal cortex (MFC). It is unknown precisely how the cerebellum influences the frontal cortex and what type of information is reciprocally relayed between these two regions. A subset of neurons in the cerebellar dentate nuclei, or the homologous lateral cerebellar nuclei (LCN) in rodents, express D1 dopamine receptors (D1DRs) and may play a role in cognitive processes. We investigated how pharmacologically blocking LCN D1DRs influences performance in an interval timing task and impacts neuronal activity in the frontal cortex. Interval timing requires executive processes such as working memory, attention, and planning and is known to rely on both the frontal cortex and cerebellum. In our interval timing task, male rats indicated their estimates of the passage of a period of several seconds by making lever presses for a water reward. We have shown that a cue-evoked burst of low-frequency activity in the MFC initiates ramping activity (i.e., monotonic increases or decreases of firing rate over time) in single MFC neurons. These patterns of activity are associated with successful interval timing performance. Here we explored how blocking right LCN D1DRs with the D1DR antagonist SCH23390 influences timing performance and neural activity in the contralateral (left) MFC. Our results indicate that blocking LCN D1DRs impaired some measures of interval timing performance. Additionally, ramping activity of MFC single units was significantly attenuated. These data provide insight into how catecholamines in the LCN may drive MFC neuronal dynamics to influence cognitive function.

Changes in otoacoustic emissions during selective auditory and visual attention.

Previous studies have demonstrated that the otoacoustic emissions (OAEs) measured during behavioral tasks can have different magnitudes when subjects are attending selectively or not attending. The implication is that the cognitive and perceptual demands of a task can affect the first neural stage of auditory processing-the sensory receptors themselves. However, the directions of the reported attentional effects have been inconsistent, the magnitudes of the observed differences typically have been small, and comparisons across studies have been made difficult by significant procedural differences. In this study, a nonlinear version of the stimulus-frequency OAE (SFOAE), called the nSFOAE, was used to measure cochlear responses from human subjects while they simultaneously performed behavioral tasks requiring selective auditory attention (dichotic or diotic listening), selective visual attention, or relative inattention. Within subjects, the differences in nSFOAE magnitude between inattention and attention conditions were about 2-3 dB for both auditory and visual modalities, and the effect sizes for the differences typically were large for both nSFOAE magnitude and phase. These results reveal that the cochlear efferent reflex is differentially active during selective attention and inattention, for both auditory and visual tasks, although they do not reveal how attention is improved when efferent activity is greater.

Properties of a nonlinear version of the stimulus-frequency otoacoustic emission.

A procedure for extracting the nonlinear component of the stimulus-frequency otoacoustic emission (SFOAE) is described. This nSFOAE measures the amount by which the cochlear response deviates from linear additivity when the input stimulus is doubled in amplitude. When a 4.0-kHz tone was presented alone, the magnitude of the nSFOAE response remained essentially constant throughout the 400-ms duration of the tone; response magnitude did increase monotonically with increasing tone level. When a wideband noise was presented alone, nSFOAE magnitude increased over the initial 100- to 200-ms portion of the 400-ms duration of the noise. When the tone and the wideband noise were presented simultaneously, nSFOAE magnitude decreased momentarily, then increased substantially for about the first 100 ms and then remained strong for the remainder of the presentation. Manipulations of the noise bandwidth revealed that the low-frequency components were primarily responsible for this rising, dynamic response; no rising segment was seen with bandpass or highpass noise. The rising, dynamic nSFOAE response is likely attributable to activation of the medial olivocochlear efferent system. This perstimulatory emission appears to have the potential to provide information about the earliest stages of auditory processing for stimuli commonly used in psychoacoustical tasks.

Overshoot using very short signal delays.

The detectability of a 10-ms tone masked by a 400-ms wideband noise was measured as a function of the delay in the onset of the tone compared to the onset of the noise burst. Unlike most studies like this on auditory overshoot, special attention was given to signal delays between 0 and 45 ms. Nine well-practiced subjects were tested using an adaptive psychophysical procedure in which the level of the masking noise was adjusted to estimate 79% correct detections. Tones of both 3.0 and 4.0 kHz, at different levels, were used as signals. For the subjects showing overshoot, detectability remained approximately constant for at least 20-30 ms of signal delay, and then detectability began to improve gradually toward its maximum at about 150-200 ms. That is, there was a "hesitation" prior to detectability beginning to improve, and the duration of this hesitation was similar to that seen in physiological measurements of the medial olivocochlear (MOC) system. This result provides further support for the hypothesis that the MOC efferent system makes a major contribution to overshoot in simultaneous masking.

Selective attention reduces physiological noise in the external ear canals of humans. I: auditory attention.

In this study, a nonlinear version of the stimulus-frequency OAE (SFOAE), called the nSFOAE, was used to measure cochlear responses from human subjects while they simultaneously performed behavioral tasks requiring, or not requiring, selective auditory attention. Appended to each stimulus presentation, and included in the calculation of each nSFOAE response, was a 30-ms silent period that was used to estimate the level of the inherent physiological noise in the ear canals of our subjects during each behavioral condition. Physiological-noise magnitudes were higher (noisier) for all subjects in the inattention task, and lower (quieter) in the selective auditory-attention tasks. These noise measures initially were made at the frequency of our nSFOAE probe tone (4.0 kHz), but the same attention effects also were observed across a wide range of frequencies. We attribute the observed differences in physiological-noise magnitudes between the inattention and attention conditions to different levels of efferent activation associated with the differing attentional demands of the behavioral tasks. One hypothesis is that when the attentional demand is relatively great, efferent activation is relatively high, and a decrease in the gain of the cochlear amplifier leads to lower-amplitude cochlear activity, and thus a smaller measure of noise from the ear.

Selective attention reduces physiological noise in the external ear canals of humans. II: visual attention.

Human subjects performed in several behavioral conditions requiring, or not requiring, selective attention to visual stimuli. Specifically, the attentional task was to recognize strings of digits that had been presented visually. A nonlinear version of the stimulus-frequency otoacoustic emission (SFOAE), called the nSFOAE, was collected during the visual presentation of the digits. The segment of the physiological response discussed here occurred during brief silent periods immediately following the SFOAE-evoking stimuli. For all subjects tested, the physiological-noise magnitudes were substantially weaker (less noisy) during the tasks requiring the most visual attention. Effect sizes for the differences were >2.0. Our interpretation is that cortico-olivo influences adjusted the magnitude of efferent activation during the SFOAE-evoking stimulation depending upon the attention task in effect, and then that magnitude of efferent activation persisted throughout the silent period where it also modulated the physiological noise present. Because the results were highly similar to those obtained when the behavioral conditions involved auditory attention, similar mechanisms appear to operate both across modalities and within modalities. Supplementary measurements revealed that the efferent activation was spectrally global, as it was for auditory attention.

Overshoot measured physiologically and psychophysically in the same human ears.

A nonlinear version of the stimulus-frequency otoacoustic emission (SFOAE) was measured using stimulus waveforms similar to those used for behavioral overshoot. Behaviorally, the seven listeners were as much as 11 dB worse at detecting a brief tonal signal (4.0 kHz, 10 ms in duration) when it occurred soon after the onset of a wideband masking noise (0.1-6.0 kHz; 400 ms in duration) than when it was delayed by about 200 ms, and the nonlinear SFOAE measure exhibited a similar effect. When either lowpass (0.1-3.8 kHz) or bandpass noise (3.8-4.2 kHz) was used instead of the wideband noise, the physiological and behavioral measures again were similar. When a highpass noise (4.2-6.0 kHz) was used, the physiological and behavioral measures both showed no overshoot-like effect for five of the subjects. The physiological response to the tone decayed slowly after the termination of the noise, much like the time course of resetting for behavioral overshoot. One subject exhibited no overshoot behaviorally even though his cochlear responses were like those of the other subjects. Overall, the evidence suggests that some basic characteristics of overshoot are obligatory consequences of cochlear function, as modulated by the olivocochlear efferent system.