- Transfer and Distribution of Cantharidin within Selected Members of Blister Beetles (Coleoptera: Meloidae) and Its Probable Importance in Sexual Behaviour (2004)
- Cantharidin (C10H12O4) is a repellent against a wide variety of predators and is known to be produced by blister beetles (Coleoptera: Meloidae) and oedemerid beetles (Col: Oedemeridae) where it is found in hemolymph and various tissues. Males of blister beetles transfer large pockets of cantharidin along with sperm to the female during copulation. Cantharidin movement through different parts of male and female genitalia was surveyed to get a better time sequence of the pharmacodynamic of this compound. Since cantharidin titre in meloid beetles highly depends on the age, sex and mating record of the individuals, deuterated cantharidin (D2C) was introduced into meloids’ body by injection, mixed with artificial diet or floating the beetles’ internal organs in Ringer and D2C solution. All samples were analysed by GC-MS, but because of the approximate coelution of cantharidin and D2C, their exact measurement via normal MS seemed to be very difficult. Therefore, MRM (Multiple Reaction Monitoring) method for cantharidin/D2C separation and independent quantitation was developed for the first time. Cantharidin pharmacodynamic in male of studied species could be quite different over a period of time. As the accessory glands absorb high amount of D2C in short term, they will eventually accumulate less amount than the testis. Confirming the previous studies, it has been concluded that post-farnesyl steps in the cantharidin pathway may occur entirely in the male’s body outside the reproductive system but the ultimate product is transported into male internal genitalia via membrane of the accessory glands which should be permeable to the chemical. It transfers afterwards to epididimis and the vas deferens in higher volume comparing to the other organs of the male genitalia and finally absorbed by the testis. In females, cantharidin is first absorbed by spermatophoral receptacle in high volume while at the same time goes through ovary and is distributed upon eggs; by the way the up-taken volume by ovaries remains considerably lower than the receptacle. By time these two organs stopped accumulating of cantharidin, whereas the bursa copulatrix starts incorporating the gift actively. This reserved amount should be mainly supplied by the receptacle and ovaries directly, so that the internal way of the terpene transfer remains as the main transport way. Analysis of the eggs laid by D2C injected females of Hycleus polymorphus demonstrated that the marker has been transferred from female to the eggs. It was found that D2C had been also incorporated into the triungulin larvae; however the titre decreased significantly from egg to larva. There is a good correlation between density of cuticular pores and the cantharidin/D2C level of the scape and pedicel antennomeres of the male specimens of Cyaneolytta sp. Light microscopy of semi thin cross sections of male scape and pedicel indicates that there are plenty of canal shape structures which have been stretched from the antennal hemolymph to the antennomere surface where cuticular pores situated. These structures can be uni- or multicellular tubules which bring the circulating cantharidin of hemolymph into the surface where later on released via the opening of cuticular pores. Tracing the head capsule and antennal segments of those Cyaneolytta females which were injected by D2C shows that the marker is found mostly in the two first antennal segments. Similarly, a higher concentration of the cuticular pores was also observed on the scape and pedicel. Tubular cells or any other carrying structures such as tyloids were found neither in cross sectioning of female antennomeres nor on integument surface. We suppose a natural system has been evolved in females of genus Cyaneolytta which let them select males with abundant reservoir of cantharidin prior to any mating behaviour. Males and females have antennation during which males antennomeres as well as cantharidin containing pores which are located on the 1st and 2nd antennomeres come into direct contact with the female antennae and thus release cantharidin on its surface. So, the porous area of male antennae can be considered as cantharidin release structures (CRS). In contrast, female porous area of antennae is apparently regarded as multiporous gustatory sensillae (MGS) which means a chemoreceptor to recognize the cantharidin titre of male sexual partner. We hereby refer to cantharidin as a precopulatory sex pheromone and support the idea who suggests cantharidin role in close range sexual selection.