Yannick Wurm's public defense on 18 december 2009

1. SOCIOGENETICS OF FIRE ANTS
Yannick Wurm
supervised by Laurent Keller
December 18th. 2009

3. © Alex Wild & others

4. © Alex Wild & others

5. © Alex Wild & others

6. © Alex Wild & others

7. © Alex Wild & others

8. © Alex Wild & others

9. © Alex Wild & others

10. © Alex Wild & others

11. © Alex Wild & others

12. © Alex Wild & others

13. © Alex Wild & others

14. © Alex Wild & others

16. Coolest thing about ants: social life

17. Coolest thing about ants: social life
•key to their success

18. Coolest thing about ants: social life
•key to their success
•diverse & sophisticated

19. © National Geographic

20. © National Geographic

21. © National Geographic

22. © BBC

23. © ameisenforum.de

24. © ameisenforum.de

25. © ameisenforum.de

26. Solenopsis invicta fire ant
Workers
Queen

29. Solenopsis invicta fire ants are
a big problem!

30. Solenopsis invicta fire ants are
a big problem!

31. Solenopsis invicta fire ants are
a big problem!
a great model system!

40. After 1 to 5 days

41. Over the next weeks

42. Over the next weeks

43. Over the next weeks

44. Over the next weeks

45. Over the next weeks

46. Over the next weeks

47. Over the next weeks

48. Over the next weeks

49. Over the next weeks

50. Over the next weeks

51. Over the next weeks

52. Main questions

53. Main questions
What changes occur within the young queens after
orphaning?

54. Main questions
What changes occur within the young queens after
orphaning?
How do workers choose queens?

55. Molecular tools for ant research

56. Molecular tools for ant research
DNA? RNA? WTF?
Ant gene discovery
Microarrays

57. DNA is made up of As, Cs, Gs & Ts
DNA is stored in chromosomes

58. DNA is made up of As, Cs, Gs & Ts
DNA is stored in chromosomes

59. DNA is made up of As, Cs, Gs & Ts
DNA is stored in chromosomes

60. DNA is made up of As, Cs, Gs & Ts
DNA is stored in chromosomes

61. DNA is made up of As, Cs, Gs & Ts
DNA is stored in chromosomes

62. DNA is made up of As, Cs, Gs & Ts
DNA is stored in chromosomes
RNA indicates
gene activity

63. Fire ant gene discovery

64. Extraction
Fire ant gene discovery
Fire ants

65. Extraction
Fire ant gene discovery
+
Fire ants

66. Extraction
Fire ant gene discovery
+ +
Fire ants

67. Extraction
Fire ant gene discovery
+ +
Fire ants RNA

68. Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
RNA

69. Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

70. Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

71. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

72. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer
CAGATAGGCGATTACA
GATTACACGTGGATGC

73. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer
CAGATAGGCGATTACA
TTACACGTGGATGC

74. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer
CAGATAGGCGATTACA
GATTACACGTGGATGC

75. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer
CAGATAGGCGATTACA
GATTACACGTGGATGC

76. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

77. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

78. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

79. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

80. Assembly Sequencing Extraction
Fire ant gene discovery
+ +
Fire ants RNA
+
RNA Sequencer

81. Fire ant gene discovery
Assembly:
is complex because 10,000s of sequences

82. Fire ant gene discovery
Assembly:
is complex because 10,000s of sequences
Annotation:
inferring functions by comparing to sequences from
from other organisms

83. Fire ant gene discovery
Assembly:
is complex because 10,000s of sequences
Annotation:
inferring functions by comparing to sequences from
from other organisms
Fourmidable:
process for assembly & annotation
server for ant genetic & genomic data
Wurm et al (2009)

84. Microarrays measure gene activity
gene A gene B gene C
Wang et al (2007)

85. Microarrays measure gene activity
RNA RNA
gene A gene B gene C
Wang et al (2007)

86. Microarrays measure gene activity
RNA RNA
gene A gene B gene C
Wang et al (2007)

87. Microarrays measure gene activity
RNA RNA
gene A gene B gene C
Wang et al (2007)

88. Microarrays measure gene activity
RNA RNA
gene A gene B gene C
Wang et al (2007)

89. Microarrays measure gene activity
RNA RNA
gene A gene B gene C
Wang et al (2007)

90. Main questions
What changes occur within the young
queens after orphaning?
How do workers choose queens?

91. Gene activity after orphaning

92. Gene activity after orphaning
Field colonies Brought into lab

93. Gene activity after orphaning
Field colonies Brought into lab
Orphaning simulated
×15 simulations per colony
× 8 replicated colonies

94. Gene activity after orphaning
Field colonies Brought into lab
Orphaning simulated
Collected for RNA
0h 6h 24h
×15 simulations per colony 5 5 5
× 8 replicated colonies

95. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Wurm et al (accepted)

96. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
Wurm et al (accepted)

97. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
significant activity changes for 297 genes
Wurm et al (accepted)

98. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
significant activity changes for 297 genes
• most after 24h
Wurm et al (accepted)

99. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
significant activity changes for 297 genes
• most after 24h
• Increased activity:
Wurm et al (accepted)

100. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
significant activity changes for 297 genes
• most after 24h
• Increased activity:
• proteasomal genes
Wurm et al (accepted)

101. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
significant activity changes for 297 genes
• most after 24h
• Increased activity:
• proteasomal genes
• genes for reproductive development
Wurm et al (accepted)

102. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
significant activity changes for 297 genes
• most after 24h
• Increased activity:
• proteasomal genes
• genes for reproductive development
• Decreased activity:
Wurm et al (accepted)

103. Gene activity after orphaning
RNA labelled & hybridized to microarray of 10,000 genes
Analysis: 10,0000 linear models
significant activity changes for 297 genes
• most after 24h
• Increased activity:
• proteasomal genes
• genes for reproductive development
• Decreased activity:
• detoxification genes (GSTs, Cytochrome P450s)
Wurm et al (accepted)

104. JH: Juvenile Hormone
relative
level
0h 6h 24h Time since orphaning
0h 6h 24h

105. JH: Juvenile Hormone
relative
level
JH-degradation genes
(esterases & epoxide hydrolases)
0h 6h 24h Time since orphaning
0h 6h 24h

106. JH: Juvenile Hormone
relative
level
JH-degradation genes
(esterases & epoxide hydrolases)
0h 6h 24h Time since orphaning
JH suppressed by
degradation
0h 6h 24h before orphaning

107. Main questions
What changes occur within the young queens after
orphaning?
How do workers choose queens?

109. Competition, anyone?

110. Competition, anyone?

111. Competition, anyone?

112. Competition, anyone?

113. Competition, anyone?

114. Competition, anyone?

117. Which queen is chosen?
Known factor:
weight ≈ fertility [Fletcher & Blum, Science 1981 &1983]
vs.

118. Which queen is chosen?
Known factor:
weight ≈ fertility [Fletcher & Blum, Science 1981 &1983]
vs.
Factors we investigate:
speed of wing loss
gene activity

119. Field colonies Brought into lab
17×

120. Field colonies Brought into lab
17×
10× per
colony:

121. 17 Colonies
Time to
wing loss

122. 17 Colonies ranked
Time to
wing loss

123. 17 Colonies ranked & paired
Time to
wing loss

124. 17 Colonies ranked & paired
Time to
wing loss
• 8 “fast-slow” pairs

125. Competition setup
For each “fast-slow” pair (8 pairs total):
10 from 10 from
fast colony slow colony
Unrelated, orphaned workers

126. Competition setup
For each “fast-slow” pair (8 pairs total):
10 from 10 from
fast colony slow colony
Unrelated, orphaned workers
3 weeks later: genotyped & dissected surviving queens

127. Start of Competition Setup
Number of 10
fast
5
0
-5
Number of -10
slow A B C D E F G H
8 Competition pairs

128. Alive after 3 weeks
Number of 8
live
4
0
8 Competition pairs
-4
-8
A B C D E F G H

129. Alive after 3 weeks
Number of 8
live
4
0
8 Competition pairs
-4
-8
Prediction: moreB
A C from fast G H
D E F colonies
less from slow colonies

130. Alive after 3 weeks
Number of 8
fast
4
0
-4
Number of
slow -8
A B C D E F G H
Wurm et al (in prep)

131. Alive after 3 weeks
Higher survival of
Number of 8 from faster
fast colonies(binomial GLM likelihood
ratio test p=0.009)
4
0
-4
Number of
slow -8
A B C D E F G H
Wurm et al (in prep)

132. Alive after 3 weeks
Higher survival of
Number of 8 from faster
fast colonies(binomial GLM likelihood
ratio test p=0.009)
4
0
-4
Number of
slow -8
A B C D E F G H
Winner origin: × × Wurm et al (in prep)

133. Alive after 3 weeks
Higher survival of
Number of 8 from faster
fast colonies(binomial GLM likelihood
ratio test p=0.009)
4
0
-4
Number of But not whole story...
slow -8
A B C D E F G H
Winner origin: × × Wurm et al (in prep)

134. Gene activity analysis
Are there differences in gene activity?

135. Gene activity analysis
Are there differences in gene activity?
1. between queens from faster & slower colonies?

136. Gene activity analysis
Are there differences in gene activity?
1. between queens from faster & slower colonies?
2. between queens from winning & losing colonies?

137. Gene activity analysis
Are there differences in gene activity?
1. between queens from faster & slower colonies?
2. between queens from winning & losing colonies?
Collected
5× 24h after Microarray
orphaning
×16 colonies

138. Gene activity analysis results
Rank-based analysis (topGO/KS Elim test;
Benjamini Hochberg 95 correction) Wurm et al (in prep)

139. Gene activity analysis results
Queens from fast wing-losing queen colonies have higher:
mitochondrial activity p=10-5
transcriptional activity p=0.04
biosynthetic process activity p=0.04
Rank-based analysis (topGO/KS Elim test;
Benjamini Hochberg 95 correction) Wurm et al (in prep)

140. Gene activity analysis results
Queens from fast wing-losing queen colonies have higher:
mitochondrial activity p=10-5
transcriptional activity p=0.04
biosynthetic process activity p=0.04
Queens from winning colonies have higher:
mitochondrial activity p=0.01
organ development activity p=0.02
anatomical structure morphogenesis p=0.03
cell differentiation p=0.04
Rank-based analysis (topGO/KS Elim test;
Benjamini Hochberg 95 correction) Wurm et al (in prep)

141. Summary of answers
What changes occur within the young queens after
orphaning?
How do workers choose queens?

142. Summary of answers
What changes occur within the young queens after
orphaning?
Within 24 hours:
How do workers choose queens?

143. Summary of answers
What changes occur within the young queens after
orphaning?
Within 24 hours:
•up-regulation of proteosomal & reproductive activity
How do workers choose queens?

144. Summary of answers
What changes occur within the young queens after
orphaning?
Within 24 hours:
•up-regulation of proteosomal & reproductive activity
•down-regulation of detoxification genes
How do workers choose queens?

145. Summary of answers
What changes occur within the young queens after
orphaning?
Within 24 hours:
•up-regulation of proteosomal & reproductive activity
•down-regulation of detoxification genes
•degradation of Juvenile Hormone ceases
How do workers choose queens?

146. Summary of answers
What changes occur within the young queens after
orphaning?
Within 24 hours:
•up-regulation of proteosomal & reproductive activity
•down-regulation of detoxification genes
•degradation of Juvenile Hormone ceases
How do workers choose queens?
Choice linked to:

147. Summary of answers
What changes occur within the young queens after
orphaning?
Within 24 hours:
•up-regulation of proteosomal & reproductive activity
•down-regulation of detoxification genes
•degradation of Juvenile Hormone ceases
How do workers choose queens?
Choice linked to:
•speed of wing shedding

148. Summary of answers
What changes occur within the young queens after
orphaning?
Within 24 hours:
•up-regulation of proteosomal & reproductive activity
•down-regulation of detoxification genes
•degradation of Juvenile Hormone ceases
How do workers choose queens?
Choice linked to:
•speed of wing shedding
•activity of development, morphogenesis & differentiation genes

149. Present and future of ant sociogenetics

150. Present and future of ant sociogenetics
Tools for ant sociogenetics

151. Present and future of ant sociogenetics
Tools for ant sociogenetics
We gained insight into the molecular changes underlying
competition between queens

152. Present and future of ant sociogenetics
Tools for ant sociogenetics
We gained insight into the molecular changes underlying
competition between queens
Exciting things still to come:

153. Present and future of ant sociogenetics
Tools for ant sociogenetics
We gained insight into the molecular changes underlying
competition between queens
Exciting things still to come:
• molecular evolution studies
(impacts special lifestyles...)

154. Present and future of ant sociogenetics
Tools for ant sociogenetics
We gained insight into the molecular changes underlying
competition between queens
Exciting things still to come:
• molecular evolution studies
(impacts special lifestyles...)
• functional testing of genes
(involved in social decisions...)

155. THANKS TO....

156. THANKS TO....

157. THANKS TO....

158. THANKS TO....
Collaborators
Michael, Miguel, Dewayne Shoemaker, Ken Ross,
Vital-IT (Laurent Falquet, Ioannis, Oksana, Chris Iseli)
Those who helped computationally
Darlene Goldstein, Frederic Schütz, Volker Flegel, Marc R-R,
Julien, Fred Ricci
Those who helped in the lab
Chris LaMendola, Francoise D,
Those who make things work
France Pham, Franck, Blaise, Jerome N, Laelia...

159. Classement de la rencontre
POINTS POSITION JOUEURS POINTS
17* 11 Michaël N. 6
en 16 12 May 5
14 13 Julien 4
13 14 Dietrich 3
12 15 Michaël M. 2
ne 11 16 Alan 1
10
ophe 9
8
7

163. The sixty-five genes with the GO annotation mitochondrial inner
membrane (orange) rank high according to p-values of differential
expression between queens from faster and slower colonies (all other
genes are in dark violet)
1 1000 2000 3000 4000 5000 6000
Genes sorted by p!value from most (gene 1) to least differently expressed between queens
from faster and slower colonies (gene 6116)

166. Dismembering an unwanted virgin queen

167. •a
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