DOI: 10.5061/dryad.dv41ns25d
title: Data from: Experimental heatwaves reduce the effectiveness of ejaculates at occupying female reproductive tracts in a model insect

description: Globally, heatwaves have become more common, with hazardous consequences
 on biological processes. Recent research using a model insect (Tribolium
 castaneum) found that 5—day experimental heatwave conditions damaged
 several aspects of male reproductive biology, while females remained
 unaffected. However, females’ reproductive fitness may still be impacted,
 as insects can typically store sperm from multiple males in specialised
 organs for prolonged periods. Consequently, using males which produce
 sperm with green fluorescent protein (GFP)-tagged sperm nuclei, we
 visualised in vivo whether thermal stress affects the ejaculate occupancy
 across female storage sites under two scenarios; i) increasing time since
 insemination, and ii) in the presence of defending competitor sperm. We
 reconfirmed that sperm from heatwave-exposed males sired fewer offspring
 with previously mated females, and provide new evidence for in vivo
 distributions of heat stress-exposed males’ sperm. Sperm from
 heatwave-exposed males occupied a smaller area across the females’ storage
 sites, at lower densities. Overall, sperm occupancy decreased with time
 since insemination. However, heat stress-exposed males’ sperm were not
 lost at a relatively faster rate. The sperm from the first male to mate
 dominated the long-term storage site. However, the offensive ability of
 heat stress-exposed males’ sperm to penetrate this site was similar to
 controls.
description: Experimental approaches using lab maintained populations which
 was used to design and explore sperm performance within the reproductive
 tracts of female insects.
description: # Data from: Experimental heatwaves reduce the effectiveness of ejaculates
 at occupying female reproductive tracts in a model insect
 [https://doi.org/10.5061/dryad.dv41ns25d](https://doi.org/10.5061/dryad.dv41ns25d) ## Description of the data and file structure 3 files overall; two .csv files that are linked to the findings of this study, and a Rscript file that denotes the formal analysis ## Filename: gfpspermpaperdata copy Is the RScript that denotes the codes for analysing the data within the MS. All package information and models for interpretation of the data is in the file. Raw data/R file: General Key: Experiment description R file and associated figures Csv file Dimensions column 1 column 2 column 3 etc. ## Filename: heatwavegfpoffspringproduction A .csv file calculating the proportion of paternity share Offspring counts (number of adults, counted) Reproductive fitness of sperm in competition after male heat stress gfpspermpaperdata.R heatwavegfpoffspringproduction.csv 'data.frame': 37 obs. of 4 variables: Proppaternity: num 0.789 1 0.321 0.527 1 ... proportion of offspring sired by GFP male (vs. Rd male) [Counts to proportions] Heatwavetemp : Factor w/ 2 levels "30","41": 1 1 1 1 11 ... 5 days of control 30oC conditions or 42oC heat stress [°C] Repfitsuccess: int 15 75 43 29 75 0 71 66 51 75 ... number of offspring sired by GFP male [Counts] Repfitfail : int 4 0 91 26 0 156 0 23 27 77 ... number of offspring sired by Rd male [Counts] ## Filename: heatwavegfpspermdistribution A .csv file calculating the proportion of GFP; green fluorescent sperm present in the female reproductive tract after matings Sperm distribution in female tract after 5-day male heat stress gfpspermpaperdata.R heatwavegfpspermdistribution.csv 'data.frame': 190 obs. of 18 variables: Column.number : int 2 56 57 58 59 107 108 109 110 111 ... row number replicate : Factor w/ 190 levels "CC1","CC10","CC11",..: 1 2 3 4 5 6 7 8 9 10 ... GFP male ID [No Units] heat.treatment : Factor w/ 2 levels "control","heat": 1 1 1 1 1 1 1 1 1 1 ... GFP male 5 days of control 30oC conditions or 42oC heat stress [°C] competition.treatment : Factor w/ 2 levels "single","competition": 2 2 2 2 2 2 2 2 2 2 ... Prior mating status of female; presence/absence of sperm competition [No Units] time.post.mating.hrs : Factor w/ 2 levels "0.5","24": 1 1 1 1 1 1 1 1 1 1 ... Time between GFP mating opportunity and female freezing [Time in Hours] composite.treatment : Factor w/ 8 levels "CC0.5","CC24",..: 1 1 1 1 1 1 1 1 1 1 ... All treatments (heat, competition and time) as single factor [No Units] x10.total.tract.presence : int 1 1 1 1 1 1 1 1 1 1 ... GFP sperm visible in the female bursa. 1= yes 2 = no; sperm spread [No Units, Binomial] x10.total.tract.mean.intensity : num 6988 9412 10167 8250 8494 ... Mean pixel intensity of GFP in bursa; sperm density [Pixel intensity] x10.total.tract.median.intensity: int 7458 9853 10809 9495 12809 1822 10460 7129 10332 10674 ... Median pixel intensity of GFP in bursa; sperm density [Pixel intensity] x10.total.tract.gfp.perc.cover : num 50 87 90 72 60 35 82 51 88 87 ... % female bursa containing GFP sperm [Pixel intensity] x10.total.tract.empty.perc.cover: num 50 13 10 28 40 65 18 49 12 13 ... % female bursa not containing GFP sperm [Pixel intensity] x20.sperma.tract.presence : int 0 1 1 1 1 1 1 1 1 0 ... GFP sperm visible in the female spermatheca. 1= yes 2 = no; sperm spread [No Units, Binomial] x20.sperma.mean.intensity : num 239 3764 3341 1302 4252 ... Mean pixel intensity of GFP in spermatheca; sperm density [Pixel intensity] x20.sperma.median.intensity : int 148 2776 2893 657 4129 117 3067 104 1066 432 ... Median pixel intensity of GFP in spermatheca; sperm density [Pixel intensity] x20.sperma.gfp.perc.cover : num 0 39 22 6 60 0 56 0 1 0 ... % female spermatheca containing GFP sperm [Percentage cover of GFP] x20.sperma.empty.perc.cover : num 100 61 78 94 40 100 44 100 99 100 ... % female spermatheca not containing GFP sperm [Percentage cover of GFP] Cbind.x10.perc.cover : num [1:190, 1:2] 50 87 90 72 60 35 82 51 88 87 ... paired variable from GFP percentage cover in bursa; sperm spread [the ‘cbind’ function allowed us to calculate percentage GFP sperm cover]~NOTE, IT IS NOT A VARIABLE PRESENT IN THE DATA SUBMITTED Cbind.x20.perc.cover : num [1:190, 1:2] 0 39 22 6 60 0 56 0 1 0 ... paired variable from GFP percentage cover in spermatheca; sperm spread [this function allowed us to calculate percentage GFP sperm cover]~NOTE, IT IS NOT A VARIABLE PRESENT IN THE DATA SUBMITTED