MEPS 244:179-189 (2002)  -  doi:10.3354/meps244179

Novel behavioural assay and partial purification of a female-derived sex pheromone in Carcinus maenas

J. D. Hardege1,*, A. Jennings1, D. Hayden1, C. T. Müller1, D. Pascoe1, M. G. Bentley2,**, A. S. Clare3,**

1Cardiff School of Biosciences, Cardiff University, Park Place, PO Box 915, Cardiff CF1 3TL, United Kingdom
2Gatty Marine Laboratory, University of St Andrews, St Andrews KY16 8LB, United Kingdom
3Marine Biological Association of the United Kingdom, The Laboratory, The Hoe, Citadel Hill, Plymouth PL1 2PB, United Kingdom
Present addresses: *Department of Biological Sciences, Hull University, Cottingham Road, Hull HU6 7RX, United Kingdom. Email: **School of Marine Sciences, Ridley Building, University of Newcastle, Newcastle upon Tyne NE1 7RU, United Kingdom

ABSTRACT: Experiments were conducted to test the hypothesis that females of the shore crab Carcinus maenas release a sex pheromone to induce the pre-copula behaviour leading to the formation of mating pairs. In the novel behavioural assay, a positive response to a female-derived signal involves Œhomosexual¹ behaviour: a pre-copula male moves towards another pre-copula male that has been Œexposed¹ to the substance(s) under investigation. The assay male grasps the Œpheromone-treated¹ male (the Œpseudo-female¹) to test the hardness of the cuticle with its chelipeds, and invariably proceeds to manipulate the pseudo-female beneath its abdomen as in pre-copula (Œhomosexual pair formation¹). Control males show an agonistic behaviour towards each other. Male shore crabs will also attempt to pair with non-crab-like objects, such as stones, if these are conditioned with female pheromone. This demonstrates that male mate selection in Carcinus maenas is based primarily on the detection of female odour compounds. Urine collected from the urinary bladder of pre-copula females, as well as Œculture water¹ obtained from these females, elicited a positive response, whereas similar samples collected from inter-moult females failed to induce a sexual response in males. Preliminary characterisation of the Œwaterborne cue(s)¹ revealed the sex pheromone(s) to be smaller than 1000 Da molecular weight, sparingly soluble in organic solvents, lyophilisable, and chemically distinct from 20-hydroxyecdysone (crustecdysone) and arthropodin. Crustecdysone showed no biological activity in the assay and was not detectable in the bioactive HPLC fraction. Although this does not completely rule out a steroidal character for the sex pheromone, LC-MS analyses utilising electrospray and atmospheric pressure chemical ionisation (APCI) did not show a steroid in the bioactive fraction.

KEY WORDS: Sex pheromone · Shore crab · Bioassay · Carcinus maenas · 20-hydroxyecdysone

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