Name: operant conditioning task to measure song preference in zebra finches
Uploaded: 2019-12-26T15:00:00
Description: Watch this Scientific Journal Video about Operant Conditioning Task to Measure Song Preference in Zebra Finches Video at JoVE.com

We thank Petra Grutzik for construction of the behavioral cage, Dr. Thomas Borowski for help designing the experiment, and Dr. Stephanie White for her generous support. Funding from an HHMI undergraduate summer research grant (HHMI #30052007536 to D.S. and L.H).

All experiments were approved by the Institutional Care and Use Committee of the W.M. Keck Science Department, in accordance to NIH guidelines. All animals used were adults (&#62;90 days post hatch).
1. Construction of Operant Conditioning Chamber
<ol>
	<li>Build a testing cage composed of two identical chambers, 15 x 15 x 17 inch3 each (Figure 1A). Make a small door (as an entrance to each chamber) on the side of the cage near the pe...

<ol>
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J., Wang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neurobiology+of+pair+bonding.">The neurobiology of pair bonding.</a> Nature Neuroscience. 7 (10), 1048-1054 (2004).</li><li>Aragona, B. J., Liu, Y., Curtis, J. T., Stephan, F. K., Wang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+critical+role+for+nucleus+accumbens+dopamine+in+partner-preference+formation+in+male+prairie+voles.">A critical role for nucleus accumbens dopamine in partner-preference formation in male prairie voles.</a> Journal of Neuroscience. 23 (8), 3483-3490 (2003).</li><li>Lippert, R. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxytocin+antagonist+treatments+alter+the+formation+of+pair+relationships+in+zebra+finches+of+both+sexes.">Oxytocin antagonist treatments alter the formation of pair relationships in zebra finches of both sexes.</a> Hormones and Behavior. 62 (2), 113-119 (2012).</li><li>Adkins-Regan, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroendocrinology+of+social+behavior.">Neuroendocrinology of social behavior.</a> Ilar Journal. 50 (1), 5-14 (2009).</li><li>Goodson, J. L., Kabelik, D., Kelly, A. M., Rinaldi, J., Klatt, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Midbrain+dopamine+neurons+reflect+affiliation+phenotypes+in+finches+and+are+tightly+coupled+to+courtship.">Midbrain dopamine neurons reflect affiliation phenotypes in finches and are tightly coupled to courtship.</a> Proceedings of the National Academy of Sciences the United States of America. 106 (21), 8737-8742 (2009).</li><li>Iwasaki, M., Poulsen, T. M., Oka, K., Hessler, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexually+dimorphic+activation+of+dopaminergic+areas+depends+on+affiliation+during+courtship+and+pair+formation.">Sexually dimorphic activation of dopaminergic areas depends on affiliation during courtship and pair formation.</a> Frontiers in Behavioral Neuroscience. 8 (210), (2014).</li><li>Gess, A., Schneider, D. M., Vyas, A., Woolley, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+auditory+recognition+training+and+testing.">Automated auditory recognition training and testing.</a> Animal Behavior. 82 (2), 285-293 (2011).</li><li>Woolley, S. C., Doupe, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+context-induced+song+variation+affects+female+behavior+and+gene+expression.">Social context-induced song variation affects female behavior and gene expression.</a> PLoS Biology. 6 (3), 62 (2008).</li><li>Day, N. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=D2+dopamine+receptor+activation+induces+female+preference+for+male+song+in+the+monogamous+zebra+finch.">D2 dopamine receptor activation induces female preference for male song in the monogamous zebra finch.</a> Journal of Experimental Biology. 222, (2019).</li><li>Hahn, A. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discrimination+of+male+black-capped+chickadee+songs:+relationship+between+acoustic+preference+and+performance+accuracy.">Discrimination of male black-capped chickadee songs: relationship between acoustic preference and performance accuracy.</a> Animal Behaviour. 126, 107-121 (2017).</li><li>Leit&#227;o, A., ten Cate, C., Riebel, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Within-song+complexity+in+a+songbird+is+meaningful+to+both+male+and+female+receivers.">Within-song complexity in a songbird is meaningful to both male and female receivers.</a> Animal Behaviour. 71 (6), 1289-1296 (2006).</li><li>Day, N. F., Hobbs, T. G., Heston, J. B., White, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+Critical+Period+Learning:+Striatal+FoxP2+Affects+the+Active+Maintenance+of+Learned+Vocalizations+in+Adulthood.">Beyond Critical Period Learning: Striatal FoxP2 Affects the Active Maintenance of Learned Vocalizations in Adulthood.</a> eNeuro. 6 (2), (2019).</li><li>Chen, Y., Clark, O., Woolley, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Courtship+song+preferences+in+female+zebra+finches+are+shaped+by+developmental+auditory+experience.">Courtship song preferences in female zebra finches are shaped by developmental auditory experience.</a> Proceedings of the Royal Society B: Biological Sciences. 284 (1855), 20170054 (2017).</li><li>Heston, J. B., White, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavior-linked+FoxP2+regulation+enables+zebra+finch+vocal+learning.">Behavior-linked FoxP2 regulation enables zebra finch vocal learning.</a> Journal of Neuroscience. 35 (7), 2885-2894 (2015).</li><li>Adkins-Regan, E., Tomaszycki, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monogamy+on+the+fast+track.">Monogamy on the fast track.</a> Biological Letters. 3 (6), 617-619 (2007).</li><li>Young, R. 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We describe a method to test the song preference of zebra finches. We used this assay to test the preference of a paired female for her partner's song. In addition, the assay was used to test the effect of dopamine on induction of song preference in naive females. This method is relatively inexpensive and was designed and used by undergraduate students, making it an excellent training tool for students. Several other studies have tested song preference in several song species, making this method useful to many investigat...

An erratum was issued for:&#160;<a href="https://www.jove.com/video/60590/operant-conditioning-task-to-measure-song-preference-in-zebra-finches">Operant Conditioning Task to Measure Song Preference in Zebra Finches</a>. An affiliation&#160;was updated.
One of the affiliations was updated from:
W.M. Keck Science Department, Clarmont McKenna College &#8211; Pitzer College &#8211; Scripps College
to:
W.M. Keck Science Department, Claremont McKenna College &#8211; Pitzer College &#8211; Scripps College

An erratum was issued for:&#160;<a href="https://www.jove.com/video/60590/operant-conditioning-task-to-measure-song-preference-in-zebra-finches">Operant Conditioning Task to Measure Song Preference in Zebra Finches</a>. An affiliation&#160;was updated.

Erratum: Operant Conditioning Task to Measure Song Preference in Zebra Finches

One of the fundamental questions in biology is how animals form affiliative bonds. In particular, what are the neural mechanisms by which these bonds are made and are these basic mechanisms conserved across vertebrate species? The prairie vole has given clues to some of the neurotransmitter systems that are important for pair-bond formation1,2,3. In particular, dopamine acting through receptors in the nucleus accumbens can induce partner preference in both male and female voles4,5. It is unclear whether the general principles underlying partner preference formation and maintenance are evolutionarily conserved.
Monogamy is more common in birds than in mammals6. Therefore, comparing mechanisms of affiliative behavior in birds to other species is critical to understanding conserved neural foundations7,8,9. In general, across many species of songbirds, male song is thought to serve as an honest indicator of male fitness10,11. Male zebra finch songbirds sing to attract mates and influence the formation of monogamous pairs12,13. Thus, song can be used to determine partner preference in these birds.
The mechanisms by which females form a preference for their partner's song is unknown. Mesotocin, the avian homologue of oxytocin, appears to play a role in pair-bond formation in finches14,15. In addition, dopamine has also been shown to play a role in pair-bond formation9,16,17,18. For example, dopamine levels are higher in the nucleus accumbens of paired versus non-paired finches9.
To study the role of neurotransmitter systems on pair-bond formation and maintenance, we measured song preference using an operant testing paradigm equipped with infrared sensors to trigger song playbacks19. The ratio of song playbacks identified the female's preference for a male's song. Prior to the operant conditioning task, each female was isolated for up to 48 h in an anechoic chamber with her partner for 'paired females' or with an unfamiliar male, for 'unpaired females'. To test the effect of dopamine on song preference, unpaired females were treated twice with a D2R dopamine agonist while isolated with the unfamiliar male. This behavioral paradigm is based on previous studies19,20 and is amenable to research by undergraduate students.

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operant conditioning task to measure song preference in zebra finches

An operant conditioning paradigm is used to test the song preference of female zebra finches. Finches are placed in a two-chambered cage with a connecting opening and indicate their preference for a song by landing on a perch within each chamber. By interrupting the infrared beam from a photoelectric sensor above each perch, the bird activates the playback of a song through a speaker located on each side of the cage. Freely available software is used to trigger the song playback from each perch. To determine the song preference of each animal, her chamber preference is first identified by triggering no song playback when she lands on each perch. This chamber preference is then compared to her song preference. A minimum activity threshold is set to ensure the preference is real. Using this method, we show that paired females prefer the song of their partner. This method was used to understand the contribution of dopamine to the formation and maintenance of song preference.

Following the protocol, we found that paired females preferred their partner's song (Figure 2A). There was a significant difference between the side chamber preference during silence to that during song playback (t-test corrected for multiple comparisons; p = 0.004; t = 3.35, df = 16). Thus, the female preferentially triggered the song of her partner in comparison to the song of an unfamiliar male.
Females that were paired with an unfamiliar male ...

Watch this Scientific Journal Video about Operant Conditioning Task to Measure Song Preference in Zebra Finches Video at JoVE.com

Operant Conditioning Task to Measure Song Preference in Zebra Finches Video (Video) | JoVE

We describe a technique to evaluate song preference in zebra finches. Females are placed in a two-chambered cage and song preference is measured by the number of times she triggers the playback of one song by landing on a perch within one chamber, compared with triggering a different song in the second chamber. Perch landings are counted using infrared sensors.

Operant Conditioning Task to Measure Song Preference in Zebra Finches

An operant conditioning paradigm is used to test the song preference of female zebra finches. Finches are placed in a two-chambered cage with a connecting ...

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