[en] The song control nucleus HVC of songbirds has emerged as a widespread model system to study adult neurogenesis and the factors that modulate the incorporation of new neurons, including seasonal state, sex differences or sex steroid hormone concentrations. However, the specific function of these new neurons born in adulthood remains poorly understood. We implemented a new procedure based on focal X-ray irradiation to deplete neural progenitors in the ventricular zone adjacent to HVC and study the functional consequences. A 23 Gy dose depleted by more than 50 percent the incorporation of BrdU in neural progenitors, a depletion that was confirmed by a significant decrease in doublecortin positive neurons. This depletion of neurogenesis significantly increased the variability of testosterone-induced songs in females and decreased their bandwidth. Expression of the immediate early gene ZENK in secondary auditory areas of the telencephalon that respond to song was also inhibited. These data provide evidence that new neurons in HVC play a role in both song production and perception and that X-ray focal irradiation represents an excellent tool to advance our understanding of adult neurogenesis.
Research Center/Unit :
GIGA Neurosciences-Neuroendocrinology - ULiège
Disciplines :
Neurosciences & behavior
Author, co-author :
Chiver, Ioana ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques
Dos Santos, Ednei B; GIGA Neurosciences, University of Liege, 15 Avenue Hippocrate, 4000, Liège, Belgium
NINDS - National Institute of Neurological Disorders and Stroke F.R.S.-FNRS - Fonds de la Recherche Scientifique
Funding text :
This work was supported by a Grant from the National Institute of Neurological Disorders and Stroke (Grant RO1NS104008) to G.F.B., J.B., and C.A.C. We thank Pr. Robert Dooling and Ed. Smith, department of Psychology, University of Maryland in College Park for providing the MATLAB script used to analyze canary song. C.A.C. is F.R.S.-FNRS Research Director.
Gage, F. H., Kemperman, G. & Song, H. Adult Neurogenesis (Cold Spring Harbor Laboratory Press, 2007).
Kemperman, G. Adult Neurogenesis. Stem Cells and Neuronal Development in the Adult Brain (Oxford University Press, 2011).
Brenowitz, E. A. & Larson, T. A. Neurogenesis in the adult avian song-control system. Cold Spring Harbor Perspect. Biol. 7, a019000. 10.1101/cshperspect.a019000 (2015). DOI: 10.1101/cshperspect.a019000
Kirn, J. R. & Nottebohm, F. Direct evidence for loss and replacement of projection neurons in adult canary brain. J. Neurosci. 13, 1654–1663 (1993). DOI: 10.1523/JNEUROSCI.13-04-01654.1993
Kirn, J., O’Loughlin, B., Kasparian, S. & Nottebohm, F. Cell death and neuronal recruitment in the high vocal center of adult male canaries are temporally related to changes in song. Proc. Natl. Acad. Sci. USA 91, 7844–7848 (1994). DOI: 10.1073/pnas.91.17.7844
Goldman, S. A. Adult neurogenesis: From canaries to the clinic. J. Neurobiol. 36, 267–286 (1998). DOI: 10.1002/(SICI)1097-4695(199808)36:2<267::AID-NEU12>3.0.CO;2-B
Nottebohm, F. Neuronal replacement in adulthood. Ann. N.Y. Acad. Sci. 457, 143–161 (1985). DOI: 10.1111/j.1749-6632.1985.tb20803.x
Nottebohm, F. Neuroscience of Birdsong (eds. Zeigler, H.P. & Marler, P.). 425–448 (Cambridge University Press, 2008).
Balthazart, J. & Ball, G. F. Endocrine and social regulation of adult neurogenesis in songbirds. Front. Neuroendocrinol. 41, 3–22. 10.1016/j.yfrne.2016.03.003 (2016). DOI: 10.1016/j.yfrne.2016.03.003
Nottebohm, F., Nottebohm, M. E., Crane, L. A. & Wingfield, J. C. Seasonal changes in gonadal hormone levels of adult male canaries and their relation to song. Behav. Neural Biol. 47, 197–211 (1987). DOI: 10.1016/S0163-1047(87)90327-X
Alvarez-Buylla, A., Theelen, M. & Nottebohm, F. Birth of projection neurons in the higher vocal center of the canary forebrain before, during, and after song learning. Proc. Natl. Acad. Sci. USA 85, 8722–8726 (1988). DOI: 10.1073/pnas.85.22.8722
Rasika, S., Nottebohm, F. & Alvarez-Buylla, A. Testosterone increases the recruitment and/or survival of new high vocal center neurons in adult female canaries. Proc. Natl. Acad. Sci. USA 91, 7854–7858 (1994). DOI: 10.1073/pnas.91.17.7854
Barker, J. M., Ball, G. F. & Balthazart, J. Anatomically discrete sex differences and enhancement by testosterone of cell proliferation in the telencephalic ventricle zone of the adult canary brain. J. Chem. Neuroanat. 55, 1–8. 10.1016/j.jchemneu.2013.10.005 (2014). DOI: 10.1016/j.jchemneu.2013.10.005
Nottebohm, F. A brain for all seasons: Cyclical anatomical changes in song-control nuclei of the canary brain. Science 214, 1368–1370 (1981). DOI: 10.1126/science.7313697
Bernard, D. J., Eens, M. & Ball, G. F. Age- and behavior-related variation in volumes of song control nuclei in male European starlings. J. Neurobiol. 30, 329–339 (1996). DOI: 10.1002/(SICI)1097-4695(199607)30:3<329::AID-NEU2>3.0.CO;2-6
Brenowitz, E., Lent, K. & Kroodsma, D. E. Brain space for learned song in birds develops independently of song learning. J. Neurosci. 15, 6281 (1995). DOI: 10.1523/JNEUROSCI.15-09-06281.1995
Smith, G. T., Brenowitz, E. A. & Wingfield, J. C. Roles of photoperiod and testosterone in seasonal plasticity of the avian song control system. J. Neurobiol. 32, 426–442 (1997). DOI: 10.1002/(SICI)1097-4695(199704)32:4<426::AID-NEU6>3.0.CO;2-8
Ball, G. F., Riters, L. V. & Balthazart, J. Neuroendocrinology of song behavior and avian brain plasticity: Multiple sites of action of sex steroid hormones. Front. Neuroendocrinol. 23, 137–178 (2002). DOI: 10.1006/frne.2002.0230
Cohen, R. E., Macedo-Lima, M., Miller, K. E. & Brenowitz, E. A. Adult neurogenesis leads to the functional reconstruction of a telencephalic neural circuit. J. Neurosci. 36, 8947–8956. 10.1523/JNEUROSCI.0553-16.2016 (2016). DOI: 10.1523/JNEUROSCI.0553-16.2016
Pytte, C. L. Adult neurogenesis in the songbird: Region-specific contributions of new neurons to behavioral plasticity and stability. Brain Behav. Evol. 87, 191–204. 10.1159/000447048 (2016). DOI: 10.1159/000447048
Scharff, C., Kirn, J. R., Grossman, M., Macklis, J. D. & Nottebohm, F. Targeted neuronal death affects neuronal replacement and vocal behavior in adult songbirds. Neuron 25, 481–492 (2000). DOI: 10.1016/S0896-6273(00)80910-1
Hall, Z. J., Delaney, S. & Sherry, D. F. Inhibition of cell proliferation in black-capped chickadees suggests a role for neurogenesis in spatial learning. Dev. Neurobiol. 74, 1002–1010. 10.1002/dneu.22180 (2014). DOI: 10.1002/dneu.22180
Chen, G. & Cheng, M. F. Inhibition of lesion-induced neurogenesis impaired behavioral recovery in adult ring doves. Behav. Brain Res. 177, 358–363. 10.1016/j.bbr.2006.11.041 (2007). DOI: 10.1016/j.bbr.2006.11.041
Yang, C. F. et al. Sexually dimorphic neurons in the ventromedial hypothalamus govern mating in both sexes and aggression in males. Cell 153, 896–909. 10.1016/j.cell.2013.04.017 (2013). DOI: 10.1016/j.cell.2013.04.017
Hellier, V. et al. Female sexual behavior in mice is controlled by kisspeptin neurons. Nat. Commun. 9, 400. 10.1038/s41467-017-02797-2 (2018). DOI: 10.1038/s41467-017-02797-2
Johnston, S. et al. AAV ablates neurogenesis in the adult murine hippocampus. Elife 10.7554/eLife.59291 (2021). DOI: 10.7554/eLife.59291
Wojtowicz, J. M. Irradiation as an experimental tool in studies of adult neurogenesis. Hippocampus 16, 261–266. 10.1002/hipo.20158 (2006). DOI: 10.1002/hipo.20158
Lazarini, F. et al. Cellular and behavioral effects of cranial irradiation of the subventricular zone in adult mice. PLoS ONE 4, e7017. 10.1371/journal.pone.0007017 (2009). DOI: 10.1371/journal.pone.0007017
Manda, K., Ueno, M. & Anzai, K. Cranial irradiation-induced inhibition of neurogenesis in hippocampal dentate gyrus of adult mice: Attenuation by melatonin pretreatment. J. Pineal Res. 46, 71–78. 10.1111/j.1600-079X.2008.00632.x (2009). DOI: 10.1111/j.1600-079X.2008.00632.x
Ford, E. C. et al. Localized CT-guided irradiation inhibits neurogenesis in specific regions of the adult mouse brain. Radiat. Res. 175, 774–783. 10.1667/RR2214.1 (2011). DOI: 10.1667/RR2214.1
Achanta, P. et al. Subventricular zone localized irradiation affects the generation of proliferating neural precursor cells and the migration of neuroblasts. Stem Cells 30, 2548–2560. 10.1002/stem.1214 (2012). DOI: 10.1002/stem.1214
Kirn, J. R., Alvarez-Buylla, A. & Nottebohm, F. Production and survival of projection neurons in a forebrain vocal center of adult male canaries. J. Neurosci. 11, 1756–1762 (1991). DOI: 10.1523/JNEUROSCI.11-06-01756.1991
Kirn, J. R., Fishman, Y., Sasportas, K., Alvarez-Buylla, A. & Nottebohm, F. Fate of new neurons in adult canary high vocal center during the first 30 days after their formation. J. Comp. Neurol. 411, 487–494 (1999). DOI: 10.1002/(SICI)1096-9861(19990830)411:3<487::AID-CNE10>3.0.CO;2-M
Nottebohm, F. Testosterone triggers growth of brain vocal control nuclei in adult female canaries. Brain Res. 189, 429–436 (1980). DOI: 10.1016/0006-8993(80)90102-X
Cornez, G. et al. Testosterone stimulates perineuronal nets development around parvalbumin cells in the adult canary brain in parallel with song crystallization. Hormones Behav. 119, 104643. 10.1016/j.yhbeh.2019.104643 (2020). DOI: 10.1016/j.yhbeh.2019.104643
Madison, F. N., Rouse, M. L. Jr., Balthazart, J. & Ball, G. F. Reversing song behavior phenotype: Testosterone driven induction of singing and measures of song quality in adult male and female canaries (Serinus canaria). Gen. Comp. Endocrinol. 215, 61–75. 10.1016/j.ygcen.2014.09.008 (2015). DOI: 10.1016/j.ygcen.2014.09.008
Shevchouk, O. T., Ghorbanpoor, S., Ball, G. F., Cornil, C. A. & Balthazart, J. Testosterone-induced neuroendocrine changes in the medial preoptic area precede song activation and plasticity in song control nuclei of female canaries. Eur. J. Neurosci. 45, 886–900. 10.1111/ejn.13530 (2017). DOI: 10.1111/ejn.13530
Dos Santos, E. B., Ball, G. F., Cornil, C. A. & Balthazart, J. Treatment with androgens plus estrogens cannot reverse sex differences in song and the song control nuclei in adult canaries. Hormones Behav. 143, 105197. 10.1016/j.yhbeh.2022.105197 (2022). DOI: 10.1016/j.yhbeh.2022.105197
Cornez, G. et al. Perineuronal nets in HVC and plasticity in male canary song. PLoS ONE 16, e0252560. 10.1371/journal.pone.0252560 (2021). DOI: 10.1371/journal.pone.0252560
Shevchouk, O. T. et al. Behavioral evidence for sex steroids hypersensitivity in castrated male canaries. Horm. Behav. 103, 80–96. 10.1016/j.yhbeh.2018.06.004 (2018). DOI: 10.1016/j.yhbeh.2018.06.004
Tchernichovski, O., Nottebohm, F., Ho, C. E., Pesaran, B. & Mitra, P. P. A procedure for an automated measurement of song similarity. Anim. Behav. 59, 1167–1176 (2000). DOI: 10.1006/anbe.1999.1416
Tchernichovski, O., Eisenberg-Edidin, S. & Jarvis, E. D. Balanced imitation sustains song culture in zebra finches. Nat. Commun. 12, 2562. 10.1038/s41467-021-22852-3 (2021). DOI: 10.1038/s41467-021-22852-3
Balthazart, J., Boseret, G., Konkle, A. T., Hurley, L. L. & Ball, G. F. Doublecortin as a marker of adult neuroplasticity in the canary song control nucleus HVC. Eur. J. Neurosci. 27, 801–817. 10.1111/j.1460-9568.2008.06059.x (2008). DOI: 10.1111/j.1460-9568.2008.06059.x
Boseret, G., Ball, G. F. & Balthazart, J. The microtubule-associated protein doublecortin is broadly expressed in the telencephalon of adult canaries. J. Chem. Neuroanat. 33, 140–154. 10.1016/j.jchemneu.2007.02.002 (2007). DOI: 10.1016/j.jchemneu.2007.02.002
Mello, C. V., Vicario, D. S. & Clayton, D. F. Song presentation induces gene expression in the songbird forebrain. Proc. Natl. Acad. Sci. USA 89, 6818–6822 (1992). DOI: 10.1073/pnas.89.15.6818
Ball, G. F., Tlemcani, O. & Balthazart, J. Induction of the Zenk protein after sexual interactions in male Japanese quail. NeuroReport 8, 2965–2970. 10.1097/00001756-199709080-00032 (1997). DOI: 10.1097/00001756-199709080-00032
Mello, C. V. & Ribeiro, S. ZENK protein regulation by song in the brain of songbirds. J. Comp. Neurol. 393, 426–438 (1998). DOI: 10.1002/(SICI)1096-9861(19980420)393:4<426::AID-CNE3>3.0.CO;2-2
Maney, D. L. & Pinaud, R. Estradiol-dependent modulation of auditory processing and selectivity in songbirds. Front. Neuroendocrinol. 32, 287–302. 10.1016/j.yfrne.2010.12.002 (2011). DOI: 10.1016/j.yfrne.2010.12.002
Balthazart, J. & Ball, G. F. Endogenous versus exogenous markers of adult neurogenesis in canaries and other birds: Advantages and disadvantages. J. Comp. Neurol. 522, 4100–4120. 10.1002/cne.23661 (2014). DOI: 10.1002/cne.23661
Barker, J. M., Charlier, T. D., Ball, G. F. & Balthazart, J. A new method for in vitro detection of bromodeoxyuridine in serum: A proof of concept in a songbird species, the canary. PLoS ONE 8, e63692. 10.1371/journal.pone.0063692 (2013). DOI: 10.1371/journal.pone.0063692
Haakenson, C. M., Madison, F. N. & Ball, G. F. Effects of song experience and song quality on immediate early gene expression in female canaries (Serinus canaria). Dev. Neurobiol. 79, 521–535. 10.1002/dneu.22685 (2019). DOI: 10.1002/dneu.22685
Scott, B. B. & Lois, C. Developmental origin and identity of song system neurons born during vocal learning in songbirds. J. Comp. Neurol. 502, 202–214. 10.1002/cne.21296 (2007). DOI: 10.1002/cne.21296
Vellema, M., van der Linden, A. & Gahr, M. Area-specific migration and recruitment of new neurons in the adult songbird brain. J. Comp. Neurol. 518, 1442–1459. 10.1002/cne.22281 (2010). DOI: 10.1002/cne.22281
Alvarez-Buylla, A., Theelen, M. & Nottebohm, F. Proliferation, “hot spots” in adult avian ventricular zone reveal radial cell division. Neuron 5, 101–109 (1990). DOI: 10.1016/0896-6273(90)90038-H
Cornez, G. et al. Development of perineuronal nets during ontogeny correlates with sensorimotor vocal learning in canaries. eNeuro. 10.1523/ENEURO.0361-19.2020 (2020). DOI: 10.1523/ENEURO.0361-19.2020
Alward, B. A., Balthazart, J. & Ball, G. F. Dissociable effects on birdsong of androgen signaling in cortex-like brain regions of canaries. J. Neurosci. 37, 8612–8624. 10.1523/JNEUROSCI.3371-16.2017 (2017). DOI: 10.1523/JNEUROSCI.3371-16.2017
Scotto-Lomassese, S., Rochefort, C., Nshdejan, A. & Scharff, C. HVC interneurons are not renewed in adult male zebra finches. Eur. J. Neurosci. 25, 1663–1668. 10.1111/j.1460-9568.2007.05418.x (2007). DOI: 10.1111/j.1460-9568.2007.05418.x
Nottebohm, F., Stokes, T. M. & Leonard, C. M. Central control of song in the canary, Serinus canarius. J. Comp. Neurol. 165, 457–486 (1976). DOI: 10.1002/cne.901650405
Margoliash, D. Functional organization of forebrain pathways for song production and perception. J. Neurobiol. 33, 671–693 (1997). DOI: 10.1002/(SICI)1097-4695(19971105)33:5<671::AID-NEU12>3.0.CO;2-C
Clayton, D. F. Role of gene regulation in song circuit development and song learning. J. Neurobiol. 33, 549–571 (1997). DOI: 10.1002/(SICI)1097-4695(19971105)33:5<549::AID-NEU5>3.0.CO;2-4
Ribeiro, S., Cecchi, G. A., Magnasco, M. O. & Mello, C. V. Toward a song code: Evidence for a syllabic representation in the canary brain. Neuron 21, 359–371 (1998). DOI: 10.1016/S0896-6273(00)80545-0
Bauer, E. E. et al. A synaptic basis for auditory-vocal integration in the songbird. J. Neurosci. 28, 1509–1522. 10.1523/JNEUROSCI.3838-07.2008 (2008). DOI: 10.1523/JNEUROSCI.3838-07.2008
Akutagawa, E. & Konishi, M. New brain pathways found in the vocal control system of a songbird. J. Comp. Neurol. 518, 3086–3100. 10.1002/cne.22383 (2010). DOI: 10.1002/cne.22383
Theunissen, F. E. et al. Neuroscience of Birdsong (eds. Ziegler, P. H. & Marler, P.). 157–173 (Cambridge Univerity Press, 2008).
MacDougall-Shackleton, S. A. & Ball, G. F. Comparative studies of sex differences in the song-control system of songbirds. Trends Neurosci. 22, 432–436 (1999). DOI: 10.1016/S0166-2236(99)01434-4
Odom, K. J., Hall, M. L., Riebel, K., Omland, K. E. & Langmore, N. E. Female song is widespread and ancestral in songbirds. Nat. Commun. 5, 3379. 10.1038/ncomms4379 (2014). DOI: 10.1038/ncomms4379
Del Negro, C., Gahr, M., Leboucher, G. & Kreutzer, M. The selectivity of sexual responses to song displays: effects of partial chemical lesion of the HVC in female canaries. Behav. Brain Res. 96, 151–159 (1998). DOI: 10.1016/S0166-4328(98)00009-6
Lynch, K. S., Kleitz-Nelson, H. K. & Ball, G. F. HVC lesions modify immediate early gene expression in auditory forebrain regions of female songbirds. Dev. Neurobiol. 73, 315–323. 10.1002/dneu.22062 (2013). DOI: 10.1002/dneu.22062
Tsoi, S. C. et al. Hemispheric asymmetry in new neurons in adulthood is associated with vocal learning and auditory memory. PLoS ONE 9, e108929. 10.1371/journal.pone.0108929 (2014). DOI: 10.1371/journal.pone.0108929
Aronowitz, J. V. et al. Unilateral vocal nerve resection alters neurogenesis in the avian song system in a region-specific manner. PLoS ONE 16, e0256709. 10.1371/journal.pone.0256709 (2021). DOI: 10.1371/journal.pone.0256709
