MiR-9 regulates basal ganglia-dependent developmental vocal learning and adult vocal performance in songbirds.

Zhimin Shi,Hannah Jarrell,Zhaoqian Teng,Xiaofang Zhang,Zoe Piccus,Huidi Yang,Yan Ding,Ofer Tchernichovski,Xiaoching Li

doi:10.7554/elife.29087

Abstract

miR-9 is an evolutionarily conserved miRNA that is abundantly expressed in Area X, a basal ganglia nucleus required for vocal learning in songbirds. Here, we report that overexpression of miR-9 in Area X of juvenile zebra finches impairs developmental vocal learning, resulting in a song with syllable omission, reduced similarity to the tutor song, and altered acoustic features. miR-9 overexpression in juveniles also leads to more variable song performance in adulthood, and abolishes social context-dependent modulation of song variability. We further show that these behavioral deficits are accompanied by downregulation of FoxP1 and FoxP2, genes that are known to be associated with language impairments, as well as by disruption of dopamine signaling and widespread changes in the expression of genes that are important in circuit development and functions. These findings demonstrate a vital role for miR-9 in basal ganglia function and vocal communication, suggesting that dysregulation of miR-9 in humans may contribute to language impairments and related neurodevelopmental disorders.

Highlights

Like humans developing speech and language, juvenile songbirds learn to sing by imitating the songs of an adult tutor early in life (Doupe and Kuhl, 1999; Immelmann and Hinde, 1969; Marler and Tamura, 1964; Tchernichovski et al, 2001)
We found that the expression levels of both FoxP1 and FoxP2 mRNAs were significantly downregulated in Area X injected with lenti-miR-9 compared to Area X injected with lenti-control (Figure 7A, p = 0.006 for FoxP1 and p < 0.0001 for FoxP2, n = 7), whereas no change in expression of either FoxP1 or FoxP2 mRNA was found in tissue adjacent to Area X
We found that both FoxP1 and FoxP2 protein levels were downregulated in Area X injected with the lenti-miR-9 virus compared to controls (Figure 7B, p = 0.0007 for FoxP1 and p = 0.0037 for FoxP2, n = 4)

Summary

Introduction

Like humans developing speech and language, juvenile songbirds learn to sing by imitating the songs of an adult tutor early in life (Doupe and Kuhl, 1999; Immelmann and Hinde, 1969; Marler and Tamura, 1964; Tchernichovski et al, 2001). Through thousands of trial-and-error practice sessions guided by auditory feedback, the juvenile song gradually matures into a stereotyped adult song that resembles the tutor’s song (Immelmann and Hinde, 1969; Konishi, 1965; Tchernichovski et al, 2001). Depending on the social context, adult males sing a courtship song directed toward a female (DS) or an undirected song (UDS) when singing alone (Sossinka and Bohner, 1980). These two types of songs exhibit subtle differences in acoustic features, and the DS is more stereotyped than the UDS (Hessler and Doupe, 1999; Jarvis et al, 1998; Kao and Brainard, 2006; Murugan et al, 2013)

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Results

Conclusion