Observations on the settlement and cementation of barnacle (Balanus amphitrite) cyprid larvae after artificial exposure to noradrenaline and the locations of adrenergic-like receptors

Abstract

Exploration of substrata prior to cementation and metamorphosis is a critical step in the life cycle of Cirripedes. To this end, barnacle cyprid larvae have evolved sophisticated sensory mechanisms to investigate chemical as well as topographical composition of the substratum. Since barnacles are major biofoulers, the sensory structures of Balanus amphitrite cyprids have been described in detail using ultrastructural techniques like SEM and TEM. Settlement assays have shown that cyprid settlement is deterred by adrenergic receptor agonists and antagonists, yet the exact location of these receptors on cyprid sensory structures is unknown. Here, using a fluorescent adrenergic receptor antagonist (Bodipy FL-Prazosin) with the aid of confocal microscopy we describe the putative locations of adrenergic-like receptors in B. amphitrite cyprids. Our results indicate that postaxial setae 2 and 3 of the 2nd and 3rd segments respectively, the sub-terminal setae of the 4th antennular segment and the cement gland secretory cells seem to bear the adrenergic-like receptors. Using optical and electron microscopy we further studied the effects of noradrenaline (NA), an adrenergic receptor agonist, on cyprid settlement and metamorphosis. NA treated cyprids forgo the typical settlement behavior and metamorphose into juvenile barnacles without cementing to the substratum. Cyprid-barnacle metamorphosis in NA treated cyprids is also prolonged as compared to untreated barnacles. The key role played by adrenergic-like receptors during surface sensing, cementation and metamorphosis is discussed.