A mating plug in a squid? Sneaker spermatophores can block the female sperm-storage organ in Doryteuthis plei

Males from numerous animal taxa have evolved strategies for obstructing the female genitalia with copulatory plugs, reducing the risk of sperm competition and thus resulting in an advantage in sexual selection. Several lines of evidence suggest that sperm competition is a common feature in the complex squid mating systems, which include the evolution of alternative mating tactics (consort vs. sneaker). However, mating plugs have hitherto not been reported for the group. Investigating the female sperm-storage organ (i.e., seminal receptacle, SR) of the squid Doryteuthis plei, we found cases in which everted spermatophores (i.e., spermatangia) were implanted into the SR and blocking its opening. Here, we describe this finding of "plugged spermatangia" based on microscopy analyses (histology and microCT) of SRs of females from three experimental groups (before and after recent mating and after egg release). We show that sneaker male spermatophores may block the opening of the SR, possibly functioning as temporary copulatory plugs that physically obstruct the SR. Together with previous experimental data on spermatophore functioning, our results suggest that plug efficiency is high until at least 5 h after mating, when spermatangia are turgid and full of sperm, clogging the organ's opening. After that time, plugs gradually decrease their efficiency as they lose turgidity by releasing part of their sperm content. However, one experimental female still had a plugged spermatangium blocking a major portion of the opening even after 48 h without mating. Within the context of squid mating systems and sexual selection, we hypothesize that plugged spermatangia are a sneaker strategy associated with minimizing sperm competition between sneaker males.