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. 2012 Oct;15(10):1371-3.
doi: 10.1038/nn.3218. Epub 2012 Sep 16.

Reversible switching between epigenetic states in honeybee behavioral subcastes

Brian R Herb  1 , Florian Wolschin, Kasper D Hansen, Martin J Aryee, Ben Langmead, Rafael Irizarry, Gro V Amdam, Andrew P Feinberg
Affiliations

Reversible switching between epigenetic states in honeybee behavioral subcastes

Brian R Herb et al. Nat Neurosci. 2012 Oct.

Abstract

In honeybee societies, distinct caste phenotypes are created from the same genotype, suggesting a role for epigenetics in deriving these behaviorally different phenotypes. We found no differences in DNA methylation between irreversible worker and queen castes, but substantial differences between nurses and forager subcastes. Reverting foragers back to nurses reestablished methylation levels for a majority of genes and provides, to the best of our knowledge, the first evidence in any organism of reversible epigenetic changes associated with behavior.

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Figures

Figure 1
Figure 1. DNA methylation changes found between nurses and foragers, but not between queens and workers
a, We compared newly emerged queens and workers using CHARM (n=5 per phenotype), and found no statistically significant differences. b, DNA methylation changes during nurse to forager transition and change back during forager to nurse transition (n=3 per phenotype). We found 155 DMRs associated with the nurse to forager transition, 107 DMRs associated with the forager to nurse transition, and 57 DMRs common to both lists that exhibit a nurse specific signature.
Figure 2
Figure 2. DNA methylation distinguishes nurses, foragers and reverted nurses
Two examples of CHARM DMRs. a-b, Top panels show percent methylation for both CHARM and WGBS data sets, with points representing individual samples, and the smoothed lines representing the average for the phenotype. The t-test panel displays the top 1% differentially methylated CpGs by t-test. Color of the point indicates which phenotype has greater methylation at that CpG (n=6 per phenotype). The RNAseq expression panel is a t-statistic based on the number or reads detected within the annotated exons, with the color indicating the higher expressed phenotype. The Exon junctions panel is a t-statistic based on the number or reads detected spanning the exon junctions, as predicted by the TopHat program, with the color indicating the higher expressed phenotype. Switching between higher expressed nurse and forager exon junctions is indicative of alternative splicing events. The RNA reads panels indicate the number of reads per phenotype as compiled by TopHat program (n=6 per phenotype). The bottom two panels show the CpG density, and the relative position of the gene. c. Plot of relative gene expression comparing foragers to reverted nurses. 26 genes associated with DMRs were tested for gene expression differences by real-time PCR (n=12 per phenotype). The plot depicts the difference in average log2 expression versus average difference in methylation as determined by CHARM. Correlation analysis results in a P-value of 0.001.
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