Brooding in the Chilean oyster Ostrea chilensis: unexpected complexity in the movements of brooded offspring within the mantle cavity.

Brooding in invertebrates serves to protect embryos from stressful external conditions by retaining progeny inside the female body, effectively reducing the risk of pelagic stages being exposed to predation or other environmental stressors, but with accompanying changes in pallial fluid characteristics, including reduced oxygen availability. Brooded embryos are usually immobile and often encapsulated, but in some Ostrea species the embryos move freely inside the female pallial cavity in close association with the mother's gills for as long as eight weeks. We used endoscopic techniques to characterize the circulation pattern of embryos brooded by females of the oyster, Ostrea chilensis. Progeny at embryonic and veliger stages typically circulated in established patterns that included the use of dorsal and ventral food grooves (DFG, VFG) to move anteriorly on the gills. Both embryos and veligers accumulated around the mother's palps, and remained there until an active maternal countercurrent moved them to the gill inhalant area. Both food grooves were able to move embryos, veligers, and food-particle aggregates anteriorly, but the DFG was more important in progeny transport; early embryos were moved more rapidly than veligers in the DFG. A microcirculation pattern of embryos was apparent when they were moved by gill lamellae: when they were close to the VFG, most embryos lost gill contact and "fell" down to the DFG. Those that actually reached the DFG moved anteriorly, but others came into contact with the base of the lamellae and again moved towards the VFG. The circulation pattern of the progeny appears well-suited for both cleaning them and directing them posteriorly to an area where there is more oxygen and food than in the palp region. This process for actively circulating progeny involves the feeding structures (gill and palps) and appears to be energetically costly for the female. It also interferes with feeding, which could explain the poor energy balance previously documented for brooding females of this species.

Quantitative analysis of sperm plane circular movement in the blue mussels Mytilus edulis, M. trossulus and their hybrids.

Fertilization success of free spawning organisms such as Mytilus species depends on gamete interactions. Therefore, gamete traits such as sperm movement are important for determining fertilization success in free spawning organisms. Since little is known about sperm movement pattern in Mytilus species, the purpose of this study was to investigate sperm movement pattern of blue mussel M. edulis, M. trossulus and their hybrids using computer-assisted sperm movement video analysis. Sperm of all genotypes were found to conduct circular movement in a two-dimensional plane. Furthermore, new sperm movement parameters, real time radius (R), angle change rate (θ) and the center of circular track (O(t)) were developed to verify and quantitatively describe the plane circular movement pattern using software (Image-J) that may be widely applied to sperm movement study in other organisms. Angle change rate was positively correlated to fertilization success. However, no correlation between fertilization and real time radius was detected. Although no interspecific differences were found in the radius, the F¹ (first generation) hybrid sperm had a lower angle change rate than M. edulis and M. trossulus. Published studies have shown that sperm circular movement is more prevalent in aquatic broadcast spawning species than in species with mating behavior or internal fertilization. Therefore, a two-dimensional circular movement pattern in sperm may represent a trait that increases fertilization success for broadcast spawning species by either increasing gamete interaction rate at a small scale and/or avoiding swimming further away from the eggs before sperm detects the chemoattractant gradient.

Incomplete reproductive isolation in the blue mussel (Mytilus edulis and M. trossulus) hybrid zone in the Northwest Atlantic: role of gamete interactions and larval viability.

Reproductive isolation in free-spawning organisms may involve only small changes in the gamete surface molecules that control fertilization, linking gamete incompatibility and speciation. Most studies have focused on species in which natural hybrids are absent and reproductive isolation is complete, but how gamete incompatibility evolves remains unclear. Reproductive isolation is incomplete between two sympatric mussel species (Mytilus edulis, M. trossulus) that hybridize in nature. In this study prezygotic and postzygotic components of reproductive incompatibility were examined in laboratory crosses. Conspecific crosses showed significantly greater rates of fertilization than heterospecific crosses, although some females of both species showed heterospecific gamete compatibility. The proportion of fertilized eggs developing into normal larvae was not significantly different between conspecific and heterospecific crosses, but survival of normal larvae was greater for conspecific crosses. Mixed-species sperm experiments suggested that conspecific sperm preference may further limit hybridization. The different components of reproductive incompatibility and total incompatibility varied among females of both species. Although our study has shown that partial reproductive isolation between M. edulis and M. trossulus involves both prezygotic gamete interactions and postzygotic larval survival, further research is required to determine the potential role of gamete incompatibility in the evolution of complete reproductive isolation between these species.