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| Status |
Public on Jan 01, 2009 |
| Title |
Defining Behavioral and Molecular Differences Between Summer and Migratory Monarch Butterflies |
| Organism |
Danaus plexippus |
| Experiment type |
Expression profiling by array
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| Summary |
In the fall, Eastern North American monarch butterflies (Danaus plexippus) undergo a magnificent long-range migration. In contrast to spring and summer butterflies, fall migrants are juvenile hormone deficient, which leads to reproductive arrest and increased longevity. Migrants also use a time-compensated sun compass to help them navigate in the south/southwesterly direction en route for Mexico. Central issues in this area are defining the relationship between juvenile hormone status and oriented flight, critical features that differentiate summer monarchs from fall migrants, and identifying molecular correlates of behavioral state. Here we show that increasing juvenile hormone activity to induce summer-like reproductive development in fall migrants does not alter directional flight behavior or its time-compensated orientation, as monitored in a flight simulator. Reproductive summer butterflies, in contrast, uniformly fail to exhibit directional, oriented flight. To define molecular correlates of behavioral state, we used microarray analysis of 9417 unique cDNA sequences. Gene expression profiles reveal a suite of 40 genes whose differential expression in brain correlates with oriented flight behavior in individual migrants, independent of juvenile hormone activity, thereby separating molecularly fall migrants from summer butterflies. Intriguing genes that are differentially regulated include the clock gene vrille and the locomotion-relevant tyramine beta hydroxylase gene. In addition, several differentially regulated genes (37.5% of total) are not annotated, suggesting unique functions associated with oriented flight behavior. We also identified 23 juvenile hormone-dependent genes in brain, which separate reproductive from non-reproductive monarchs; genes involved in longevity, fatty acid metabolism, and innate immunity are upregulated in non-reproductive (juvenile-hormone deficient) migrants. The results link key behavioral traits with gene expression profiles in brain that differentiate migratory from summer butterflies and thus show that seasonal changes in genomic function help define the migratory state.
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| Overall design |
A total of 40 monarch butterflies were used for the microarray analysis. Of the 40, 10 (5 male/5 female) were summer butterflies (Designated as S) and 30 were fall butterflies. The fall butterflies were further divided into three groups: 10 (5 male/5 female) were untreated (F); 10 (5 male/5 female) were treated with methoprene (M), which is a juvenile hormone analog and induces the development of reproductive organs in migrant butterflies; and 10 (5 male/5 female) were treated with vehicle only (V). We collected total brain RNA from each of the 40 butterflies. The brain RNAs were amplified and then used to probe a custom Nimblegen array that was designed to analyze the 9,417 unique cDNA sequences established in our published EST library (http://titan.biotec.uiuc.edu/cgi-bin/ESTWebsite/estima_start?seqSet=butterfly).
Our main interest is to find genes involved in migration. This includes genes regulating oriented flight behavior of the butterfly and genes that regulate reproductive status. To identify these genes, we approached the microarray data in two ways.
First, we identified the potential genes involved in oriented flight behavior using the following strategy. We compared the summer group to each of the three fall groups (untreated, methoprene-treated, and vehicle-treated) for males and for females, and looked for gene regulation patterns common among the three comparisons for each sex. Because the comparisons were done separately for males and females, and our behavioral data did not show significant sex differences in flight orientation, we focused on the common differentially regulated genes that were shared between males and females. Accordingly, we identified 40 cDNAs that were differentially regulated between summer butterflies and fall migrants, irrespective of sex.
Second, we looked for the juvenile hormone-response genes. Again, we performed sex-specific statistical analyses, and compared the summer and the fall groups, and the methoprene-treated and vehicle-treated migrants. We then screened for shared genes between the two groups for each sex. We next examined cDNAs that were differentially regulated in both males and females, to determine juvenile hormone-regulated genes involved in more global processes (e.g., longevity and fatty acid metabolism) that would not be expected to be sex-specific. We identified 23 putative juvenile hormone-response genes that were common between males and females.
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| Contributor(s) |
Zhu H, Gegear RJ, Casselnab A, Kanginakudru S, Reppert SM |
| Citation(s) |
19335876 |
| Submission date |
Dec 18, 2008 |
| Last update date |
Mar 20, 2012 |
| Contact name |
Steven Reppert |
| E-mail(s) |
steven.reppert@umassmed.edu
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| Phone |
508-856-6148
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| Fax |
508-856-6266
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| Organization name |
University of Massachusetts Medical School
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| Department |
Neurobiology
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| Street address |
364 Plantation St
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| City |
Worcester |
| State/province |
MA |
| ZIP/Postal code |
01605 |
| Country |
USA |
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| Platforms (1) |
| GPL7829 |
Umass/Reppert Danaus plexippus EST custom Nimblegen array |
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| Samples (40)
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| GSM352399 |
S-M03 Monarch Butterfly Summer Male 03 |
| GSM352401 |
S-M04 Monarch Butterfly Summer Male 04 |
| GSM352402 |
S-M07 Monarch Butterfly Summer Male 07 |
| GSM352403 |
S-M10 Monarch Butterfly Summer Male 10 |
| GSM352404 |
S-M11 Monarch Butterfly Summer Male 11 |
| GSM352405 |
S-F19 Monarch Butterfly Summer Female 19 |
| GSM352406 |
S-F20 Monarch Butterfly Summer Female 20 |
| GSM352407 |
S-F21 Monarch Butterfly Summer Female 21 |
| GSM352408 |
S-F22 Monarch Butterfly Summer Female 22 |
| GSM352409 |
S-F23 Monarch Butterfly Summer Female 23 |
| GSM352410 |
F-M07 Monarch Butterfly Fall Male 07 |
| GSM352411 |
F-M10 Monarch Butterfly Fall Male 10 |
| GSM352412 |
F-M23 Monarch Butterfly Fall Male 23 |
| GSM352413 |
F-M29 Monarch Butterfly Fall Male 29 |
| GSM352414 |
F-M32 Monarch Butterfly Fall Male 32 |
| GSM352415 |
F-F03 Monarch Butterfly Fall Female 03 |
| GSM352416 |
F-F13 Monarch Butterfly Fall Female 13 |
| GSM352417 |
F-F14 Monarch Butterfly Fall Female 14 |
| GSM352418 |
F-F18 Monarch Butterfly Fall Female 18 |
| GSM352419 |
F-F21 Monarch Butterfly Fall Female 21 |
| GSM352420 |
M-M03 Monarch Butterfly Fall Methoprene Treated Male 03 |
| GSM352421 |
M-M04 Monarch Butterfly Fall Methoprene Treated Male 04 |
| GSM352422 |
M-M08 Monarch Butterfly Fall Methoprene Treated Male 08 |
| GSM352423 |
M-M13 Monarch Butterfly Fall Methoprene Treated Male 13 |
| GSM352424 |
M-M15 Monarch Butterfly Fall Methoprene Treated Male 15 |
| GSM352425 |
M-F20 Monarch Butterfly Fall Methoprene Treated Female 20 |
| GSM352426 |
M-F23 Monarch Butterfly Fall Methoprene Treated Female 23 |
| GSM352427 |
M-F24 Monarch Butterfly Fall Methoprene Treated Female 24 |
| GSM352428 |
M-F27 Monarch Butterfly Fall Methoprene Treated Female 27 |
| GSM352445 |
M-F30 Monarch Butterfly Fall Methoprene Treated Female 30 |
| GSM352554 |
V-M10 Monarch Butterfly Fall Vehicle Treated Male 10 |
| GSM352555 |
V-M13 Monarch Butterfly Fall Vehicle Treated Male 13 |
| GSM352556 |
V-M14 Monarch Butterfly Fall Vehicle Treated Male 14 |
| GSM352557 |
V-M18 Monarch Butterfly Fall Vehicle Treated Male 18 |
| GSM352558 |
V-M19 Monarch Butterfly Fall Vehicle Treated Male 19 |
| GSM352559 |
V-F04 Monarch Butterfly Fall Vehicle Treated Female 04 |
| GSM352560 |
V-F22 Monarch Butterfly Fall Vehicle Treated Female 22 |
| GSM352561 |
V-F24 Monarch Butterfly Fall Vehicle Treated Female 24 |
| GSM352562 |
V-F27 Monarch Butterfly Fall Vehicle Treated Female 27 |
| GSM352563 |
V-F29 Monarch Butterfly Fall Vehicle Treated Female 29 |
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| Relations |
| BioProject |
PRJNA112399 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE14041_RAW.tar |
49.0 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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