Alterations in ZENK Expression in Zebra Finches Reared by a Single Parent Tareq Yousef, Jordan Fisk,...
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Transcript of Alterations in ZENK Expression in Zebra Finches Reared by a Single Parent Tareq Yousef, Jordan Fisk,...
Alterations in ZENK Expression in Zebra Finches Reared by a Single Parent
Tareq Yousef, Jordan Fisk, Tara S. Perrot, and Leslie S. PhillmoreDepartment of Psychology and Neuroscience
Introduction
Dalhousie University, Halifax, NS Canada
Conclusions
Future Directions
References
Acknowledgments
Bolhuis JJ, Zijlstra GCO, Den Boer-Visser SM, Van der Zee EA. (2000). Localized neuronal activation in the zebra finch brain is related to the strength of song learning. Proc Natl Acad Sci USA 97:2282-2285.MacDougall-Shackleton SA, Hulse SH, Ball GF. (1998). Neural bases of song preferences in female zebra finches (Taeniopygia guttata). NeuroReport 9:3047–3052.Maney D. & Pinaud R. (2011). Estradiol-dependent modulation of auditory processing and selectivity in songbirds. Frontiers in Neuroendocrinology, 32:287-302.
Results
Methods ZENK-ir - Effects of Having a Single Parent
Song Learning Song Learning:• Having a single father did not
compromise song learning in male
offspring.
• Having a single mother did not reduce
similarity to father’s song to zero,
indicating there must be a minimal
level of similarity among all male
songs.
• All birds sang consistently.
ZENK-ir in NCM and CMM:• CMM, important for conspecific
recognition in males and song
preference in females, had the
greatest ZENK-ir in birds reared
without their father in response to
hearing playback of their father’s song
for the first time. Birds reared with a
father present (biparental or single
father) were familiar with father’s song,
possibly affecting ZENK-ir.
• In NCM, rearing group had no effect on
ZENK-ir in males, but appears to affect
ZENK-ir in females.
NCM and Song Learning:• Amount of ZENK-ir was positively
correlated with how well father’s song
was copied (Bolhuis et al. 2000), but
only when birds were reared by both
parents.
Determine if other factors, such as
number of male siblings, may affect
amount of ZENK-ir in finches reared
without a father.
Finches paired and allowed to breed
Biparental Father or mother removed 4-5d post hatch
Allow birds to mature in natural nest groups
Single female broods separated from other groups with adult males
Record directed song of adult male
offspring ZENK-ir – Song Learning and Response in NCM
Song Analyses
As part of a larger project examining the effects of having a single parent in a normally biparental species, we wondered how rearing offspring with only a father or mother might affect song learning in males and neural response to father’s song in males and females.In this study we removed one parent of a brood and examined:
•How well a son copied their father’s song•Whether zenk protein immunoreactivity (ZENK-ir) to playback of
father’s song was correlated with strength of learning in males (as in Bolhuis et al. 2000)
•Whether ZENK-ir to father’s song varied in females reared with one or both parents
• Recorded father’s directed song before breeding
• Recorded offspring’s directed song after PHD 90
• Used Sound Analysis Pro (Tchernichovski) to assess:
• Similarity to father’s song (% of song copied)
• Of the song copied from father, how accurate that copy was
• Similarity to self (stability)
Sons with single fathers did not have lower song similarity or accuracy to father’s song than sons in biparental nests.Sons without fathers had lower accuracy copying father’s song, and a trend toward lower similarity (p=0.055).
Despite differences in song quality, all offspring sang their own song consistently, and were not different from fathers’ consistency.
The authors wish to thank Ashley Lockyer, Dominique Shephard, and Jaya Wadhawan for help with bird care, video recording, and project management.
*
Recording of fathers and male offspring were collected by exposing birds to live females and vocal recordings after isolating the male overnight in a sound-attenuated chamber. IHC for zenk protein (SC-189) followed standard protocols after playback of father’s song to offspring. Images captured at 20x and ROI placed as in Maney et al. 2011.
Fathers Biparental Single Mother Single Father0
10
20
30
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60
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80
90
100
Group
% S
imila
rity
Am
on
g R
en
dit
ion
s
Playback father’s song to male and female offspring Isolation overnight, 30 min playback, sac 60 min after end playback,
Record responses to playback.
Section brain tissue (coronal) and process in immunohistochemistry for zenk. Count number of labeled cells within ROI placed in NCM and CMM.
Biparental Single Mother Single Father0
10
20
30
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100
Group
% S
imila
rity
to
Fa
the
r's
So
ng
Biparental Single mother Single Father0
10
20
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50
60
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80
90
100
Group
% A
cc
ura
cy
of
No
tes
Co
pie
d
* *
Analyzing all male offspring together, there was no correlation between ZENK-ir and similarity to father’s song (Spearman’s rho (14)=0.02, p=0.29) or accuracy of copy (rho(14) = 0.04, p=0.89).
Males reared by both parents: ZENK-ir and similarity were positively but not significantly correlated (rho(4)=0.32, p=-.68); same for ZENK-ir and accuracy (rho(4)=0.95,p=0.051).
Males reared by a single mother: ZENK-ir was negatively but not significantly correlated with similarity (rho(3)=-0.50, p=0.67).
Males reared by a single father: ZENK-ir was negatively but not significantly correlated with similarity (rho(7)=-0.23, p=0.61).
Biparental Single Mother Single Father0
10
20
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50
60
70
80
90
100
Male
Female
Group
# Z
EN
K-I
R c
ells
NCMImage captured at 4x, coronal section, ROI placed within NCM.
A group x sex ANOVA showed no differences between rearing groups or between sexes. A group x sex interaction F(2,15) = 3.58, p=0.054) indicated a trend that ZENK-ir differed between males and females in the single mother group but did not differ between males and females in the other groups.
Biparental Single Mother Single Father0
10
20
30
40
50
60
70
80
Group
# Z
EN
K-I
R c
ells
**
A group x sex ANOVA showed no difference between sexes nor a group x sex interaction. However, there was a significant effect of group F(2,15)=6.59, p = 0.01; offspring reared by single mothers had greater ZENK-ir to playback of their father than birds reared with their father.
CMMImage captured at 4x, coronal section, ROI placed within CMM.
0 5 10 15 20 25 30 35 400
10
20
30
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50
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90
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R² = 0.000526485506956442
Number of ZENK-ir Cells in NCM
% S
imil
ari
ty t
o F
ath
er'
s S
on
g
0 5 10 15 20 25 30 35 400
10
20
30
40
50
60
70
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90
100
R² = 0.00171002121608044
Number of ZENK-ir Cells in NCM
Ac
cu
rac
y o
f C
op
y
0 5 10 15 20 25 30 35 400
10
20
30
40
50
60
70
80
90
100
R² = 0.0382433076513931
Number of ZENK-ir Cells in NCM
% S
imil
ari
ty t
o F
ath
er'
s S
on
g
5 10 15 20 25 30 35 400
10
20
30
40
50
60
70
80
90
100
R² = 0.0167905797980418
Number of ZENK-ir Cells in NCM
% S
imil
ari
ty t
o F
ath
er'
s S
on
g
5 10 15 20 25 30 350
10
20
30
40
50
60
70
80
90
100
R² = 0.0380518104969466
Number of ZENK-ir Cells in NCM
% S
imil
ari
ty t
o F
ath
er'
s S
on
g
Biparental Single Mother Single Father