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. 2018 Sep 12;16(9):e2006537. doi: 10.1371/journal.pbio.2006537

Fig 5. Singing experience–mediated dendritic spine pruning in RA projection neurons.

Fig 5

(A) Coinduction of RA cluster I genes (Arc, Nr4a1, Sik1, and Dusp5) after juvenile singing in glutamatergic neurons with Vglut2 (+) but not GABAergic neurons with Gad2 (+). Filled and empty arrowheads: cells that coexpressed or did not coexpress with singing activity–dependent genes and cell marker genes, respectively. Cell nuclei (blue, DAPI). Scale bar = 40 μm. Bar graphs: proportion of each subpopulation in cells that express the mRNA of RA Cluster I genes. (B) Selective induction of Arc mRNA (green) after juvenile singing in RA projection neurons. Diagram of DiI retrograde labeling (red) of RA projection neurons to nXIIts. Cell nuclei (blue, DAPI). (C) Golgi-stained RA projection neurons and RA-surrounding arcopallial neurons in juvenile (55 phd), adult (105 phd), and SP adult (101 phd at 1–2 days after release) birds. Scale bars = 5 μm (upper) and 50 μm (lower). Bar graphs: dendritic spine density of RA projection neurons and RA-surrounding arcopallial neurons in juveniles (n = 18 cells from 6 birds), adults (n = 15 cells from 5 birds), and SP adults. ***P < 0.0001, NS: p > 0, unpaired t test with Bonferroni correction. Error bars: SEM. Supporting data can be found in S5 Data. NS, not significant; phd, post hatching day; RA, robust nucleus of the arcopallium; SP, singing-prevented.