1 Function of copulatory plugs in house mice: mating behaviour and paternity 2 outcomes of rival males 3 4 Andreas Sutter1,3, Leigh W. Simmons2, Anna K. Lindholm1 and Renée C. Firman2 5 6 1Institute of Evolutionary Biology and Environmental Studies, University of Zurich, 7 Winterthurerstrasse 190, 8057 Zurich, Switzerland. 8 2Centre for Evolutionary Biology, School of Animal Biology (M092), The University of Western 9 Australia, Crawley, 6009, Australia. 10 3 E-mail:
[email protected] 11 12 Corresponding author: Andreas Sutter, IEU – Animal Behavior, Winterthurerstrasse 190, 8057 13 Zürich, Switzerland; Tel: +41 79 545 9015; email:
[email protected]; 14 15 Abbreviated title: Function of copulatory plugs 16 1 17 Abstract 18 Polyandry is widespread across animal taxa, and subjects males to intense post-copulatory sexual 19 selection which favors adaptations that enhance a male’s paternity success, either by decreasing the 20 risk of sperm competition and/or by increasing the competitiveness of the ejaculate. Copulatory 21 plugs deposited by males are thought to have evolved in the context of sperm competition. 22 However, experimental studies that assess the function of copulatory plugs remain scarce. 23 Moreover, most studies have used unnatural manipulations, such as ablating plug-producing male 24 glands or interrupting copulations. Here, we investigated whether repeated ejaculation affects plug 25 size in a mammalian model species, the house mouse. When males experience short periods of 26 sexual rest we found that plug size decreased over repeated ejaculations so that time since last 27 ejaculation can be applied as an approximation for plug size.