Songbirds possess a spontaneous ability to discriminate syntactic rules

Transcription

1 Songbirds possess a spontaneous ability to discriminate syntactic rules Kentaro Abe and Dai Watanabe Supplementary Figures 1 4 and Legends 1

2 Supplementary Fig. 1. (a) SMSD-test on a single bird (S24). The raw data of call response number over the 5 min period before and after the song change. Data of 8 trials are plotted. (b) Individual differences of the behavioral response to the song changes. The number of call responses over the 5 min periods before and after the song change are averaged for each individuals. Data of the 17 male subjects used in Fig. 1c are shown. (c) Normalized shift in call response during the stimuli change. Data of birds purchased (n = 54, 19 birds) or raised in our aviary (n = 50, 15 birds) are shown. P-values of paired-t-test of raw call counts before and after the stimuli change are indicated. * P < Data are normalized to the total call counts in 10 min (after + before) for each stimuli change. Mean ± s.e.m. 2

3 Supplementary Fig. 2. Effects of the systematic modification of syllable sequences in SMSD-test. Since we were unaware of whether natural songs of Bengalese finches contain any syllable ordering rules, shared within the community, the initial syllable modification had to be somewhat arbitrary. We initially created 3 sequence modified songs, SEQ1 3 (a), concentrating to characteristic syllable doublings in ORI songs (i.e. AA or GG). Since only the SEQ2 were discriminated among those songs (Fig. 1c d), we further created a systematic modification based on the modifications performed to SEQ2. Since SEQ2 contained two modification to ORI songs, we created SEQ2B,2C and 2D (Fig. 1g) to clarify which sequence modification are responsible for the discrimination of SEQ2. The results were shown in Fig. 1e. Based on the results, we further performed a systematic modification of syllable sequences concentrating on where to insert the GGL sequence. (SEQ2E M b,c). However, as described in the text, this systematic analysis failed to find a simple liner sequence that the birds may use to discriminate the sequence modified songs. Mean ± s.e.m. * P < 0.05, n.s. P > 0.05, paired-t-test of raw call counts before and after the stimuli change. Data from different experiment are summarized in each graph., n = 39, 16 birds., n = birds., n = 44, 16 birds. Same data of SEQ2, SEQ2B, SEQ2C and SEQ2D shown in Fig. 1f were shown. 3

4 Supplementary Fig. 3. Sonogram of songs of the isolate reared siblings (S25M, S26M, S27M) at PHD130. Two songs for each bird are shown. 4

5 Supplementary Fig. 4. Parasagital sections of the anterior nidopallium (a), and arcopallium (b), immunostained with NeuN (green) and DAPI (blue). The birds were sacrificed 2 weeks after the injection of ibotenic acid or PBS as a control. (c) Outlined drawings of the lesions in the anterior nidopallium performed to the other 8 birds used in the experiment in Fig. 5. White lines indicate the border of lesions revealed by NeuN stained images. Scale Bars, 200 m. (d) Relation of lesioned volume of the anterior nidopallium and the normalized shift in call response against SEQ2 in the SMSD-test. Lesioned volumes were calculated by outlining the lesioned area once in five sections throughout both hemispheres. Red line indicates the average volume of MAN in the both hemispheres of control birds, 0.23 ± mm 3. (e,f) Effects of lesioning of anterior nidopallium and RA to motoric behaviors. Raw counts of behavior / min (e, hop, f, call) during the initial song exposure in HAB-test are shown. No significant effects of motoric disturbances were observed in the lesioned birds. Mean ± s.e.m. n = 9, n.s. P > 0.05, one-way ANOVA. 5