Faculty Bibliography

Vocal communication is negatively affected by neurodegenerative diseases, such as Parkinson disease, and by aging. The neurological and sensorimotor mechanisms underlying voice deficits in Parkinson disease and aging are not well-understood. Rat ultrasonic vocalizations provide a unique behavioral model for studying communication deficits and the mechanisms underlying these deficits in these conditions. The purpose of this review was to examine the existing literature for methods using rat ultrasonic vocalization with regard to the primary disease pathology of Parkinson disease, dopamine denervation, and aging. Although only a small amount of papers were found for each of these topics, results suggest that both shared and unique acoustic deficits in ultrasonic vocalizations exist across conditions and that these acoustic deficits are due to changes in either dopamine signaling or denervation and in aging models changes to the nucleus ambiguus, at the level of the neuromuscular junction, and the composition of the vocal folds in the larynx. We conclude that ultrasonic vocalizations are a useful tool for studying biologic mechanisms underlying vocal communication deficits in neurodegenerative diseases and aging.

Aging affects voice production and is associated with reduced communicative ability and quality of life. Voice therapy is a critical component of treatment, but its effects on neuromuscular mechanisms are unknown. The ultrasonic vocalizations (USVs) of rats can be used to test the effects of aging and voice use on the laryngeal neuromuscular system. This study tested the hypothesis that age-related changes in the USVs of rats and laryngeal neuromuscular junctions can be reversed through vocal exercise. Young and old rats were trained for 8 weeks to increase their USVs and were compared with a no intervention group pre- and post-treatment. USV acoustics and aspects of neuromuscular junction (NMJ) morphology were measured in the thyroarytenoid muscle. Vocal training reduced or eliminated some age differences found in both USVs and NMJs. We conclude that vocal exercise may assist in mitigating age-related changes in voice characteristics and underlying neuromuscular adaptations.

OBJECTIVES/HYPOTHESIS: Singing-voice classification is often considered the cornerstone of a classical singer's identity. Traditionally, classification has been a highly subjective, nonstandardized process. As a result, misclassification of the singing voice is thought to be common, especially in young singers. Long-term average spectrum (LTAS) average is an objective measurement that could be used to classify a singer's voice. The purpose of this study was to determine the relationship of LTAS with singing-voice classification. STUDY DESIGN: Descriptive between-subject study. METHODS: Nine professional classical male singers performed the "Star Spangled Banner" in a comfortable key of their choice. LTAS was calculated for the first two phrases, the remainder of the song, and the entire song. The overall LTAS averages of each sample as well as the physiological and singing ranges were compared with self-reported singing-voice classification. RESULTS: Voice classification and overall LTAS average were moderately correlated, but the strength of the correlation varied with each sample. The strongest correlation was with the entire song. Voice classification and singing range were strongly correlated. CONCLUSIONS: LTAS remains a promising tool to aid in singing-voice classification. However, how to best use LTAS in classification remains unclear because of the influence of sample length and phonetic and pitch content on LTAS.

Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting communication abilities. (1, 2) Targeted training (speech/voice therapy) can improve specific voice deficits,(3, 4) although the underlying mechanisms of behavioral interventions are not well understood. Systematic investigation of voice deficits and therapy should consider many factors that are difficult to control in humans, such as age, home environment, age post-onset of disease, severity of disease, and medications. The method presented here uses an animal model of vocalization that allows for systematic study of how underlying sensorimotor mechanisms change with targeted voice training. The ultrasonic recording and analysis procedures outlined in this protocol are applicable to any investigation of rodent ultrasonic vocalizations. The ultrasonic vocalizations of rodents are emerging as a valuable model to investigate the neural substrates of behavior.(5-8) Both rodent and human vocalizations carry semiotic value and are produced by modifying an egressive airflow with a laryngeal constriction.(9, 10) Thus, rodent vocalizations may be a useful model to study voice deficits in a sensorimotor context. Further, rat models allow us to study the neurobiological underpinnings of recovery from deficits with targeted training. To model PD we use Long-Evans rats (Charles River Laboratories International, Inc.) and induce parkinsonism by a unilateral infusion of 7 mug of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle which causes moderate to severe degeneration of presynaptic striatal neurons (for details see Ciucci, 2010).(11, 12) For our aging model we use the Fischer 344/Brown Norway F1 (National Institute on Aging). Our primary method for eliciting vocalizations is to expose sexually-experienced male rats to sexually receptive female rats. When the male becomes interested in the female, the female is removed and the male continues to vocalize. By rewarding complex vocalizations with food or water, both the number of complex vocalizations and the rate of vocalizations can be increased (Figure 1). An ultrasonic microphone mounted above the male's home cage records the vocalizations. Recording begins after the female rat is removed to isolate the male calls. Vocalizations can be viewed in real time for training or recorded and analyzed offline. By recording and acoustically analyzing vocalizations before and after vocal training, the effects of disease and restoration of normal function with training can be assessed. This model also allows us to relate the observed behavioral (vocal) improvements to changes in the brain and neuromuscular system.

Alterations in neuromuscular junction (NMJ) structure in cranial muscles may contribute to age-related deficits in critical sensorimotor actions such as swallowing. Neuromuscular electrical stimulation (NMES) is used in swallowing therapy, but it is unclear how NMJ structure is affected or if NMJ morphology is best measured in two or three dimensions. Two- and three-dimensional measurements of NMJ morphology in the genioglossus muscle were compared in rats that had undergone 8 weeks of hypoglossal nerve stimulation vs. untreated controls. The relationship between motor endplate volume and nerve terminal volume had a mean positive slope in 90% of the young adult controls, but it was positive in only 50% of the old controls; 89% of NMES old rats had a positive slope. NMJ measurements were more accurate when measured in three dimensions. In the NMJ, aging and NMES are associated with changes in the pre- and post-synaptic relationship.

The source of ultrasonic vocalizations (USVs) produced by rats is thought to be within the larynx. The purpose of this investigation was to determine if the rat larynx is capable of producing ultrasounds with the full range of frequencies reported in vivo. Acoustic output of excised rat larynges with and without vocal fold constriction was measured. At biologically-reasonable airflow rates and pressures, only larynges with a constriction produced the full range of ultrasounds reported in vivo, providing support for the hypothesis that a constriction within the larynx is likely the source of rat USVs.