Nuptial food gifts offered by males to females at mating are shaped by sexual conflict, allowing males direct access to female physiology. However, a molecular dissection of their effect on females is rare. In decorated crickets, the male’s nuptial gift comprises part of the male’s spermatophore, the spermatophylax, which functions to deter the female from prematurely removing the sperm-containing portion, the ampulla, from her genital opening. However, ingested spermatophylax compounds and proteins contained in the ampulla could also influence female physiology and behavior to the male’s benefit. We investigated how mating per se and these two distinct routes of potential male-mediated manipulation influence the transcriptional response of females. We conducted an RNA-sequencing experiment on the gut and head tissue from females for whom consumption of nuptial food gifts and receipt of an ejaculate had been independently manipulated. In the gut tissue, we found that females not permitted to feed during mating exhibit a decreased expression of many genes, which seems to be caused by reduced gut function, but this was countered by female feeding on the spermatophylax or a sham gift. In the head tissue, we found only low numbers of differentially expressed genes, but a gene co-expression network analysis revealed that both the attachment of the ampulla and the consumption of the spermatophylax independently induce their own distinct patterns of gene expression. This study provides evidence that spermatophylax feeding alters the female post-mating transcriptomic response in decorated crickets, highlighting its potential to mediate sexual conflict in this system.

Transcriptomes were assembled with Trinity and sare deposited at DDBJ/EMBL/GenBank under the bioproject PRJNA784797. 

They were annotated using the Trinotate pipeline. Transcripts were translated into their most likely coding regions, if any, using Transdecoder. Both the resulting protein products and all original transcripts were used to find similar sequences in the Swiss-Prot protein database, using either BLASTP or BLASTX with a threshold of E ≤ 10−5. Signal peptides, transmembrane helices and protein domains were predicted using SignalP v4.1, tmhmm v2.0 and HMERR with the PFAM database, respectively. 

The results, in addition to KEGG, Eggnog, and Gene Ontology (GO) annotations were parsed by Trinotate and stored in a SQLite database. The two CSV files were exported from such SQLite databases, and include all annotations found by Trinotate.

All real-time quantitative PCR (qPCR) experiments were performed following the MIQE guidelines for qPCR experiments. For each qPCR reaction, 2 µL of cDNA was added to 10 µL of Power SYBR™ Green PCR Master Mix (Fisher #4368702), 6.8 µL of H2O, and 1.2 µL of primers at a final concentration of 300 nM. All reactions were run in duplicate on 96 well plates, using the following thermal cycling profile on a QuantStudio 3 Real-Time PCR system (Thermo Fisher Scientific): 2 minutes at 50°C, 10 minutes at 95°C, 40 cycles of (1) 15 seconds at 95°C and (2) 1 min at 60°C, and a melting curve from 95°C to 60°C. We assessed the relative expression of six target genes (Hinfp, Ubtf, Nup93, Vg2, SLC35B3 and Rassf8) in the four experimental groups (SA, A, S, V).

All data files are saved as .csv files and can be opened with any text editor.