DOI: 10.5061/dryad.m37pvmd5w
title: Dopamine mediates the pea aphid wing plasticity

description: 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.
description: 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.
description: 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.