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. 2010 Apr 30:10:120.
doi: 10.1186/1471-2148-10-120.

The transcription factor Krüppel homolog 1 is linked to hormone mediated social organization in bees

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The transcription factor Krüppel homolog 1 is linked to hormone mediated social organization in bees

Hagai Shpigler et al. BMC Evol Biol. .

Abstract

Background: Regulation of worker behavior by dominant queens or workers is a hallmark of insect societies, but the underlying molecular mechanisms and their evolutionary conservation are not well understood. Honey bee and bumble bee colonies consist of a single reproductive queen and facultatively sterile workers. The queens' influences on the workers are mediated largely via inhibition of juvenile hormone titers, which affect division of labor in honey bees and worker reproduction in bumble bees. Studies in honey bees identified a transcription factor, Krüppel-homolog 1 (Kr-h1), whose expression in worker brains is significantly downregulated in the presence of a queen or queen pheromone and higher in forager bees, making this gene an ideal candidate for examining the evolutionary conservation of socially regulated pathways in Hymenoptera.

Results: In contrast to honey bees, bumble bees foragers do not have higher Kr-h1 levels relative to nurses: in one of three colonies levels were similar in nurses and foragers, and in two colonies levels were higher in nurses. Similarly to honey bees, brain Kr-h1 levels were significantly downregulated in the presence versus absence of a queen. Furthermore, in small queenless groups, Kr-h1 levels were downregulated in subordinate workers with undeveloped ovaries relative to dominant individuals with active ovaries. Brain Kr-h1 levels were upregulated by juvenile hormone treatment relative to a vehicle control. Finally, phylogenetic analysis indicates that KR-H1 orthologs are presence across insect orders. Though this protein is highly conserved between honey bees and bumble bees, there are significant differences between orthologs of insects from different orders.

Conclusions: Our results suggest that Kr-h1 is associated with juvenile hormone mediated regulation of reproduction in bumble bees. The expression of this transcription factor is inhibited by the queen and associated with endocrine mediated regulation of social organization in two species of bees. Thus, KR-H1 may transcriptionally regulate a conserved genetic module that is part of a pathway that has been co-opted to function in social behavior, and adjusts the behavior of workers to their social environmental context.

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Figures

Figure 1
Figure 1
The influence of queen presence on brain Kr-h1 levels in workers. A. In the first experiment we compared workers from queenright colonies and workers of similar age and genotype that developed in small queenless groups. Data represent mean ± standard error of mean. The number of individual brains used in each sample is shown in the base of each bar. P-values above plots were obtained from unpaired t-test analyses. B. In the second experiment we compared queenless and queenright workers that developed in colonies with similar worker and brood populations, but differing in the presence or absence of an egg-laying queen. Sample size was 6 bees/colony. Groups with different letters are significantly different in a one-way ANOVA followed by a LSD post hoc test (P < 0.001 for both tests).
Figure 2
Figure 2
The influence of task on brain Kr-h1 levels. We collected nurses and foragers from self-supported free-flying colonies. Brain Kr-h1 transcript levels were similar in foragers and nurses from Colony A but higher in nurses in colonies B and C. Data represent mean ± standard error of the mean. The number of individual brains used in each sample is shown in the base of each bar. P-values above plots were obtained from unpaired t-test analyses.
Figure 3
Figure 3
The influence of dominance rank on brain Kr-h1 levels in groups of three queenless workers. A. Average brain Kr-h1 RNA levels for bees with different dominance rank. Data represent mean ± standard error and the sample size is indicated within bars. The P-value was obtained from a one way ANOVA. Groups with different letters are significantly different in a LSD post hoc statistical test (P < 0.05). B. The proportion of bees in each dominance rank with the highest, medium, and lowest brain Kr-h1 RNA levels. The numbers inside the bars show the number of bees fitting each category. The P-value above plot shows the result of a Chi square goodness of fit test comparing the observed values to an expected probability of 0.33 if dominance and Kr-h1 levels are independent.
Figure 4
Figure 4
Phylogenetic tree of Kr-h1 orthologues. A maximum likelihood phylogenetic tree examining the relationship between the BtKR-H1 ortholog and orthologs from 19 other insect taxa, using the conserved zinc finger region (see Additional file 1,2 Fig. S1). Maximum likelihood bootstrap values in percent (1000 replicates) are shown at each node. Branch lengths are from maximum likelihood. Insect orders are indicated to the right. The hemimetabolous insect A. pisum is set as the outgroup to the holometablous insects. B. terrestris and A. mellifera are shown within the Hymenoptera. The GenBank accession numbers and full names of all species can be found in the methods.

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