Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Jul 7;106(27):11364-9.
doi: 10.1073/pnas.0812998106. Epub 2009 Jun 18.

Discrete molecular states in the brain accompany changing responses to a vocal signal

Shu Dong  1 , Kirstin L Replogle, Linda Hasadsri, Brian S Imai, Peter M Yau, Sandra Rodriguez-Zas, Bruce R Southey, Jonathan V Sweedler, David F Clayton
Affiliations

Discrete molecular states in the brain accompany changing responses to a vocal signal

Shu Dong et al. Proc Natl Acad Sci U S A. .

Abstract

New experiences can trigger changes in gene expression in the brain. To understand this phenomenon better, we studied zebra finches hearing playbacks of birdsong. Earlier research had shown that initial playbacks of a novel song transiently increase the ZENK (ZIF-268, EGR1, NGFIA, KROX-24) mRNA in the auditory forebrain, but the response selectively habituates after repetition of the stimulus. Here, using DNA microarray analysis, we show that novel song exposure induces rapid changes in thousands of RNAs, with even more RNAs decreasing than increasing. Habituation training leads to the emergence of a different gene expression profile a day later, accompanied by loss of essentially all of the rapid "novel" molecular responses. The novel molecular profile is characterized by increases in genes involved in transcription and RNA processing and decreases in ion channels and putative noncoding RNAs. The "habituated" profile is dominated by changes in genes for mitochondrial proteins. A parallel proteomic analysis [2-dimensional difference gel electrophoresis (2D-DIGE) and sequencing by mass spectrometry] also detected changes in mitochondrial proteins, and direct enzyme assay demonstrated changes in both complexes I and IV in the habituated state. Thus a natural experience, in this case hearing the sound of birdsong, can lead to major shifts in energetics and macromolecular metabolism in higher centers in the brain.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Experimental treatment groups. Following a balanced design, each bird was placed alone in a sound isolation chamber on the afternoon of day 0. On day 1, birds in the habituated and trained-only groups heard repeated playback of the song stimulus for 3 h. On day 2, birds in the novel and habituated groups heard the song stimulus for 30 min. The initial combined microarray and proteomic analysis focused just on the first 3 groups (silence, novel, and habituated). A subsequent microarray experiment compared a separate trained-only group matched to a new habituated group collected at the same time. New cohorts representing all 4 groups were then prepared and assayed for mitochondrial enzyme activities.
Fig. 2.
Fig. 2.
Behavioral validation of treatment groups. The birds from novel and habituated groups used in the combined microarray/proteomics experiment (n = 24) were videotaped during the test song presentation period and their responses to the onset of the song stimulus were scored as described (13) (SI). Upon onset of song playback, zebra finches arrest other behaviors and sit silently for periods of seconds to many minutes as though listening intently to the song. The duration from first song onset until the resumption of other visible activities is termed “response latency.” The data were square-root transformed to approximate normality. The birds in the habituated group had significantly shorter response latencies than those in the novel group (*, P < 0.01, Student's t test). Error bars represent SEM.
Fig. 3.
Fig. 3.
Distinct gene sets define 4 major response patterns. (A) Venn diagram of differential expression. After pairwise comparisons of habituated and novel groups against the silence group, significant probes (Student's t test, FDR P < 0.05) were classified according to direction of change (up ↑ or down ↓) relative to the silence group. Probes present in multiple classes are indicated by overlaps, with the number of probes specific to each distinct subdivision indicated (see also Table S1). (B) Principal component analysis (PCA) of biological samples, using all probes on each array; axes represent the top 3 principal components (P1, P2, P3), which capture 54% of the overall variance. (C) PCA using only probes significant by ANOVA at FDR P < 0.05 (n = 4,341); the top 3 principal components capture 71% of the overall variance. Star, habituated group; circle, novel group; square, silence group; each symbol represents 1 bird. Note that birds naturally group according to treatment.
Fig. 4.
Fig. 4.
Mitochondrial complex activity analysis. Activities were measured (SI) in extracts from auditory lobule and anterior forebrain of birds in 4 treatment groups (n = 3 birds in each group): S, silence; N, novel; H, habituated; T, trained. All activities were normalized against the mean activity of the silence group within the same brain region. (A) complex IV activity; (B) complex I activity. ANOVA followed by post hoc test: **, P < 0.01; *, P < 0.05.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Clayton DF. The genomic action potential. Neurobiol Learn Mem. 2000;74:185–216. - PubMed
    1. Robinson GE, Fernald RD, Clayton DF. Genes and social behavior. Science. 2008;322(5903):896–900. - PMC - PubMed
    1. Dong S, Clayton DF. Habituation in songbirds. Neurobiol Learn Mem. 2008;92:183–188. - PMC - PubMed
    1. Mello CV, Vicario DS, Clayton DF. Song presentation induces gene expression in the songbird forebrain. Proc Natl Acad Sci USA. 1992;89:6818–6822. - PMC - PubMed
    1. Mello CV, Clayton DF. Song-induced ZENK gene expression in auditory pathways of songbird brain and its relation to the song control system. J Neurosci. 1994;14:6652–6666. - PMC - PubMed

Publication types

MeSH terms