Dopamine mediates the pea aphid wing plasticity
Data files
Apr 26, 2023 version files 715.59 MB
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5lines_count.Rdata
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annotation_for_featurecount.gff
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gene_information.xlsx
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pharmaceutical_injection_result.csv
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README.md
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Sample_Accession_number.xlsx
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v3.0_short_name.fa
Abstract
Many organisms exhibit phenotypic plasticity, in which developmental processes result in different phenotypes depending on their environmental context. We focus on the molecular mechanisms underlying that environmental response. Pea aphids (Acyrthosiphon pisum) show a wing dimorphism, in which pea aphid mothers produce winged or wingless daughters when exposed to a crowded or low-density environment, respectively. We investigated the role of dopamine in mediating this wing plasticity, motivated by a previous study that found higher dopamine titers in wingless- versus winged-producing aphid mothers. In this study, we found that manipulating dopamine levels in aphid mothers affected the number of winged offspring they produced. Specifically, asexual female adults injected with a dopamine agonist produced a lower percentage of winged offspring, while asexual females injected with a dopamine antagonist produced a higher percentage of winged offspring, matching expectations based on the titer difference. We also found that genes involved in dopamine synthesis, degradation, and signaling were not differentially expressed between wingless- and winged-producing aphids. This result indicates that titer regulation happens in a non-transcriptional manner or that we sampled non-relevant timepoints or tissue. Overall, our work emphasizes that dopamine is an important component of how organisms process information about their environments.
Methods
This data set comprises previously published and reanalyzed RNA-seq data and pharmaceutical injection result data.
In the RNAseq folder, the GenBank accession numbers for these sequences are SRR13238533 - SRR13238544, SRR2148902 - SRR2148909, and SRR21747201 - SRR21747212 (recorded in "Sample Accession Number.xlsx). Raw reads were processed using TrimGalore and FastQC (see “process_read.sh”). We trimmed adaptor sequences, filtered out poor-quality sequences (quality score cutoff 20), and discarded sequences shorter than 20nt. The resulting reads were then mapped to the pea aphid genome v3.0 using HISAT2 (see “map_read_hisat_v3.0genome.sh”, “modified genome and annotation” folder). All raw counts matrix was combined and processed into an R.data (“5lines_counts.Rdata”). Counts normalization and differential expression analysis were done using the v3.0 genome annotation with R package DESeq2 (“DE_analysis.Rmd”, “modified genome and annotation” folder). Information for dopamine-related genes investigated in this study can be found in table “gene_information.xlsx”.
In the pharmaceutical injection folder, the raw injection result table (“pharmaceutical_injection_result.csv”) and the script to analyze the visualized data (“pharmaceutical_injection_analysis.Rmd”) are included in the dataset. To generate the injection data, healthy adult females were pooled and randomly divided into injection groups. Aphids were injected with 1 μg/μL apomorphine (Sigma), 1 μg/μL flupenthixol (Fisher Scientific), or insect Ringer’s as control using a pulled glass capillary needle. Both apomorphine and flupenthixol were dissolved in Ringer’s. Glass needles were prepared using a micropipette puller (P-1000, Sutter Instrument) with the setting of pull=150 and heat=504. Needle tips were broken against a glass slide edge. Injection was done with a microinjector (PLI-10, Warner Instruments) for 0.1s, 5.7psi, injecting roughly 0.3μL of liquid. After injection, aphids were transferred back to plants with a density of 3/cage for 24h for reproduction. Injected adult females were then removed while their offspring were kept growing on plants. Offspring were categorized as winged or wingless after at least reaching the third instar stage. For each cage, the offspring’s winged percentage was calculated as the number of winged offspring divided by the total number of offspring.
Usage notes
Please open .csv and .xlsx files using excel. Use Rstudio for the .Rmd and .Rdata files. Other files can be viewed using notepad or pdf viewer.