自発対話音声に対する叫び声アノテーション

Faculty of Information and Computer Science, Chiba Institute of Technology会議名: 言語資源ワークショップ2022, 開催地: オンライン, 会期: 2022年8月30日-31日, 主催: 国立国語研究所 言語資源開発センター叫び声は突発的な感情表現を示す音声現象の一つである．先行研究では，自発対話音声に含まれる叫び声を感情表出系感動詞と区別して定義していた．しかし，先行研究の定義を基に叫び声と感情表出系感動詞のアノテーションを行っても，言語表現が似ている音声現象を音響的特性のみで区別する必要があるため，この二つの音声現象を区別することは困難であった．そこで，叫び声と感情表出系感動詞を区別するために改めて叫び声(scream) の定義を行った．また，発話の特徴と叫び声の特徴を併せ持った音声を発話と叫びの共起（shout）として区別した．これらの定義を基に自発対話音声に含まれる音声を収録した音声資料に対して叫び声アノテーションを行った．複数人でアノテーションした際の一致率算出を行って新たな定義と先行研究の定義との比較を行う．さらに，叫び声の事例をいくつか示し，自発的な叫び声がどのような音声言語現象として発せられているかについて考察する

Cofilin1 Controls Transcolumnar Plasticity in Dendritic Spines in Adult Barrel Cortex

<div><p>During sensory deprivation, the barrel cortex undergoes expansion of a functional column representing spared inputs (spared column), into the neighboring deprived columns (representing deprived inputs) which are in turn shrunk. As a result, the neurons in a deprived column simultaneously increase and decrease their responses to spared and deprived inputs, respectively. Previous studies revealed that dendritic spines are remodeled during this barrel map plasticity. Because cofilin1, a predominant regulator of actin filament turnover, governs both the expansion and shrinkage of the dendritic spine structure <i>in vitro</i>, it hypothetically regulates both responses in barrel map plasticity. However, this hypothesis remains untested. Using lentiviral vectors, we knocked down cofilin1 locally within layer 2/3 neurons in a deprived column. Cofilin1-knocked-down neurons were optogenetically labeled using channelrhodopsin-2, and electrophysiological recordings were targeted to these knocked-down neurons. We showed that cofilin1 knockdown impaired response increases to spared inputs but preserved response decreases to deprived inputs, indicating that cofilin1 dependency is dissociated in these two types of barrel map plasticity. To explore the structural basis of this dissociation, we then analyzed spine densities on deprived column dendritic branches, which were supposed to receive dense horizontal transcolumnar projections from the spared column. We found that spine number increased in a cofilin1-dependent manner selectively in the distal part of the supragranular layer, where most of the transcolumnar projections existed. Our findings suggest that cofilin1-mediated actin dynamics regulate functional map plasticity in an input-specific manner through the dendritic spine remodeling that occurs in the horizontal transcolumnar circuits. These new mechanistic insights into transcolumnar plasticity in adult rats may have a general significance for understanding reorganization of neocortical circuits that have more sophisticated columnar organization than the rodent neocortex, such as the primate neocortex.</p></div

Effects of CFL1 expression rescue on impaired experience-dependent plasticity.

<p>(A) Design of three mutant CFL1s that are resistant to miR-CFL1_1 (resCFL1s). Seven or eight nucleic acids within the target sequence of miR-CFL1_1 were mutated such that amino acid sequence did not change. (B) An <i>in vitro</i> test of CFL1 expression rescue by each resCFL1 in rat CFL1 (WT)- and miR-CFL1_1-expressing HEK293T cells. The mock group expressed only WT CFL1. <i>n</i> = 4 for all groups. resCFL1_1, <i>p</i> = 5.8 × 10^-6; resCFL1_2, <i>p</i> = 2.2 × 10^-5; resCFL1_3, <i>p</i> = 1.3 × 10^-4 versus miR-CFL1_1 group, Tukey-Kramer’s multiple comparison test. (C) An <i>in vitro</i> test of the effect of miR-CFL1_1 on resCFL1_1 expression. <i>n</i> = 3 for both groups. <i>p</i> = 0.30, <i>t</i>-test. (D) A schematic diagram of the bicistronic lentiviral vector that co-expresses resCFL1_1 and mCherry via a P2A peptide. (E) Targeted injection of Lenti-CaMKIIα-ChR2-eYFP-miR-CFL1_1 and Lenti-CaMKIIα-mCherry-P2A-resCFL1 to D2 column identified with intrinsic signal imaging induced focal expression of ChR2-eYFP and mCherry. Scale bar, 500 μm. (F) Fluorescent images of a coronal section infected with the two vectors. Scale bar, 300 μm. (G) Confocal images of an infected area showing co-expression of ChR2-eYFP and mCherry in L2/3 neurons. Scale bar, 20 μm. (H) A representative raster plot (100 trials are shown in horizontal row) and peristimulus time histogram (PSTH) of a putative ChR2+ neuron recorded from L2/3 in D2 column of the rat showed in E. (I) A representative raster plot (50 trials) and PSTH of the same neuron with (H), showing responses to D1 whisker deflections. (J) Comparison of average responses recorded from ChR2+ neurons in D2 L2/3 of deprived rats expressing miR-CFL1_1 and resCFL1 with those recorded from ChR2+ neurons in deprived rats expressing only miR-CFL1_1 and those recorded from WT deprived rats. Data of miR-CFL1_1 deprived and WT deprived groups were the same with those shown in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1002070#pbio.1002070.g003" target="_blank">Fig. 3D</a>. <i>n</i> = 17 units (from three rats) for the miR-CFL1_1+resCFL1 deprived group. *<i>p</i> = 0.0069, Tukey-Kramer’s multiple comparison test.</p

Effects of CFL1 KD on spine density during sensory deprivation.

<p>(A, B) Lentiviral vectors employed for co-expressing eGFP and miRNAs (A) and for expressing tdTomato (B). (C) The D1 and D2 barrel columns were identified via intrinsic signal optical imaging. (D) Virus injection was targeted to L2/3 of the D1 and D2 columns. (E) A representative parasagittal section showing expression of tdTomato (D1) and eGFP (D2). Scale bar, 300 μm. (F) Magnified view of an eGFP-expressing region in the parasagittal section shown in (E). Scale bar, 100 μm. (G) Confocal images of a rectangle region shown in (F). Scale bar, 20 μm. (H) A representative dendritic branch in the D2 column is shown that made a putative synaptic connection with a tdTomato+ D1 axonal bouton. Dendritic spines were counted that were localized at a distance less than 15 μm from an identified putative synaptic connection. A magnified view of the putative synaptic connection is shown in the inset. Scale bar, 10 μm. (I) Representative images of the dendritic branches within the distal portion in the D2 column. Scale bar, 10 μm. (J) Spine densities measured in the distal portion. <i>n</i> = 18, 17, 19, and 19 branch segments for miR-Neg non-deprived (ND), miR-Neg deprived (D), miR-CFL1_1 ND, and miR-CFL1_1 D groups, respectively. WT ND, <i>p</i> = 1.1 × 10^-4; miR-CFL1_1 ND, <i>p</i> = 1.8 × 10^-5; miR-CFL1_1 D, <i>p</i> = 0.0027 versus WT D group, Tukey-Kramer’s multiple comparison test. (K) Cumulative frequency histogram of spine density. WT ND, <i>p</i> = 0.037; miR-CFL1_1 ND, <i>p</i> = 3.6 × 10^-4; miR-CFL1_1 D, <i>p</i> = 3.6 × 10^-4 versus WT D group, Kolmogorov-Smirnov test with Bonferroni’s correction. (L−N) Same as (I−K) but of dendritic branches measured within the proximal portion. <i>n</i> = 14, 18, 16, and 10 branch segments for miR-Neg ND, miR-Neg D, miR-CFL1_1 ND, and miR-CFL1_1 D groups, respectively. WT ND, <i>p</i> = 0.86; miR-CFL1_1 ND, <i>p</i> = 0.97; miR-CFL1_1 D, <i>p</i> = 0.93 versus WT D group, Tukey-Kramer’s multiple comparison test.</p

Efficiency and specificity of CFL1 KD through miR-CFL1.

<p>(A) CFL1 mRNA KD efficiency of two miRNAs (miR-CFL1_1 and miR-CFL1_2) targeted against different sequences within the CFL1 gene, assessed by using CFL1-overexpressing HEK 293T-cells. CFL1 mRNA levels were normalized to those of the miR-Neg group. <i>n</i> = 3 for all groups; miR-CFL1_1, <i>p</i> = 2.7 × 10^-8; miR-CFL1_2, <i>p</i> = 2.8 × 10^-8 versus miR-Neg, Dunnett’s multiple comparison test. (B) KD efficiency of miR-CFL1_1 and miR-CFL1_2 for endogenous CFL1 protein, assessed by using PC-12 cells. “WT” (wild type) indicates PC-12 cells that were not infected with lentivirus. (C) Two neighboring coronal sections obtained from a miR-CFL1_1-expressing rat are shown, one stained with an antibody against CFL1 (left) and the other stained with an antibody against NeuN (right). The eYFP fluorescence image (middle) was obtained from the NeuN-stained section. Scale bar, 300 μm. (D) Magnified view of the rectangular region indicated in (C). Scale bar, 150 μm. (E, F) Same as (C and D), but of two neighboring coronal sections derived from a miR-Neg virus-injected rat. (G, H) Confocal images of eYFP+ or eYFP− region in a coronal section obtained from a miR-CFL1_1-expressing rat stained with antibodies against NeuN (blue) and CFL1 (red). Scale bar, 50 μm. (I) Percentage of CFL1+ cells in NeuN+ cells measured in miR-CFL1_1- or miR-Neg-expressing rats. <i>n</i> = 3 for all groups. *<i>p</i> = 0.0003, t-test with Bonferroni’s correction. (J) Effects of miR-CFL1 on mRNA expression of genes related to CFL1, assessed in PC-12 cells. <i>n</i> = 3 for all groups. miR-CFL1_1 of CFL1, <i>p</i> = 2.6 × 10^-7; miR-CFL1_2 of CFL1, <i>p</i> = 5.2 × 10^-7 versus miR-Neg, Dunnett’s multiple comparison test. (K) Effects of miR-CFL1 on expression of ADF protein in PC-12 cells. (L) Three successive coronal sections obtained from a miR-CFL1_1-expressing rat are shown, one stained with an antibody against ADF (middle) and another stained with an antibody against CFL1 (right). Scale bar, 300 μm.</p