Eugenio Carpizo-Ituarte

Role of protein kinase C, G-protein coupled receptors, and calcium flux during metamorphosis of the sea urchin Strongylocentrotus purpuratus.

Artificial inducers have been used to study signal-transduction pathways involved in metamorphosis of some marine invertebrates. However, the transduction mechanisms for echinoderms have been less explored. In the present study, participation of protein kinase C (PKC), G-protein-coupled receptors (GPCRs), and calcium has been investigated during metamorphosis of the sea urchin Stronglylocentrotus purpuratus. Competent larvae were induced with different drugs that activate (PKC and GP activators, Ca2+ ionophores, and inhibitors of Ca2+ ATPase) or inhibit (PKC, G-protein, and Ca2+ flux inhibitors) metamorphosis. Six of the compounds were effective: the PKC activators TPA and indolactam; the G-protein inhibitors suramin and guanosine; the calcium ionophore A23187, and the calcium ATPase inhibitor thapsigargin. TPA was effective at 0.001 μM; indolactam was effective at 0.001 μM. In the presence of KCl as inducer, the G-protein inhibitor suramin was effective at 10 μM and guanosine at 0.001 μM. In the presence of a bacterial film as inducer, suramin was effective at 50 μM, and guanosine inhibited metamorphosis at 1 μM. A23187 was effective at 5 and 10 μM and thapsigargin at 50 and 100 μM. Our results indicate that GPCRs, protein kinase C, and calcium participate in the metamorphosis of S. purpuratus. These elements of the transduction pathways triggered during metamorphosis may be part of a cascade of signal transduction routes that interact from induction to the end of the morphogenetic events that shape the postlarval form. In addition, according to the results obtained with G-protein inhibitors, the GPCRs may be shared between the artificial (KCl) and natural (biofilm) inducers.

Transcription and Translation Inhibitors Permit Metamorphosis up to Radiole Formation in the Serpulid Polychaete Hydroides elegans Haswell

Settlement and metamorphosis in most well-studied marine invertebrates are rapid processes, triggered by external cues. How this initial environmentally mediated response is transduced into morphogenetic events that culminate in the formation of a functional juvenile is still not well understood for any marine invertebrate. The response of larvae of the serpulid polychaete Hydroides elegans to inhibitors of mRNA and protein synthesis was examined to determine if metamorphosis requires these molecular processes. Competent larvae of H. elegans were induced to metamorphose by exposing them to a bacterial film or a 3-h pulse of 10 mM CsCl in the presence of the gene-transcription inhibitor DRB (5,6-dichloro-1-β-D-ribofuranosylbenzimidazole) or the translation inhibitor emetine. When induced to metamorphose in the presence of either inhibitor, larvae of H. elegans progressed through metamorphosis to the point at which branchial radioles start to develop. DRB and emetine inhibited the incorporation of radiolabeled uridine into RNA and radiolabeled methionine into peptides, respectively, indicating that they were effective in blocking the appropriate syntheses. Taken together, these results indicate that the induction of metamorphosis in H. elegans does not require de novo transcription or translation, and that the form of the juvenile worm is achieved in two phases. During the first phase, larvae respond to the inducer by attaching to the substratum, secreting a primary tube, resorbing the prototroch cilia, undergoing caudal elongation, and differentiating the collar; once the collar is formed, they begin secreting the secondary, calcified tube. During the second phase, the small worm develops branchial radioles and begins to grow, requiring new mRNA and protein syntheses.

Coral Reproduction in the Eastern Pacific

Sexual reproductive activity has been demonstrated in all reef-building (zooxanthellate) scleractinian corals examined from Mexico to the equatorial eastern Pacific (Galapagos Islands). Eleven of 13 species spawn gametes, six are gonochoric, three hermaphroditic, and four exhibit significant mixed sexuality (both gonochoric and hermaphroditic). Four or 30.1 %, two species each of Pocillopora and Porites, produce autotrophic ova. Porites panamensis is the only known zooxanthellate brooder. Also sexually active are the azooxanthellate scleractinian Tubastraea coccinea and the zooxanthellate hydrocoral Millepora intricata. Reproductive structures, sex ratios, age at sexual maturity, sexuality, and developmental mode have been determined from largely histological evidence. Agariciid corals, comprising more than one-third of investigated species, exhibit predominantly mixed sexual systems with sequential cosexual hermaphroditic cycles in four species. Mixed sexuality is also minimally exhibited in populations of two dominantly gonochoric species. Several eastern Pacific corals spawn mostly on lunar day 17 and 1–2 days following; however, multispecific spawning has not been observed probably because of seasonal, diel, and variable timing in spawning behavior. Factors contributing to the high fecundity of eastern Pacific corals include (1) seasonally prolonged reproductive activity, (2) small size of mature gametes allowing for production of high numbers, (3) split spawning with bimonthly gamete production in some species, (4) alternation of sex maturation in gamete development, and possibly (5) their low latitudinal location under relatively constant and high thermal conditions. Coral community persistence, reef growth and recovery are highly dependent on both sexual and asexual reproductive processes. Asexual fragmentation by physical and biotic causes is particularly important, especially for branching pocilloporid species and the fungiid coral Diaseris distorta. Asexual propagation in massive and encrusting poritid and agariciid species is also common-place, often the result of bioerosion and colony breakage by foraging reef fishes. Some research areas in need of attention are noted, for example (a) timing of spawning and the behavior of gamete release of several species, (b) life cycles of Pocillopora spp. and Millepora intricata, and (c) effects of anthropogenic stressors on eastern Pacific coral reproduction and recruitment.

The effect of filamentous turf algal removal on the development of gametes of the coral Orbicella annularis.

Macroalgae and filamentous turf algae (FTA) are abundant on degraded coral reefs, and the reproductive responses of corals may indicate sub-lethal stress under these conditions. The percentage of gametogenic stages (PGS) and the maximum diameter of eggs (MDE; or egg size) of Orbicella annularis were used to evaluate the effect of long- (7–10 months) and short-term (2.5 months) FTA removal (treatments T1 and T2, respectively) at both the beginning (May) and the end (August) of gametogenesis. Ramets (individual lobes of a colony) surrounded by FTA (T3) or crustose coralline algae (CCA; T4) were used as controls. The removal of FTA enhanced the development of gametes (i.e., a larger and higher percentage of mature gametes (PMG)) of O. annularis for T1 vs. T3 ramets in May and T1 and T2 vs. T3 ramets in August. Similar values of PGS and MDE between gametes from T3 and T4 in both May and August were unexpected because a previous study had shown that the same ramets of T4 (with higher tissue thickness, chlorophyll a cm-2 and zooxanthellae density and lower mitotic index values) were less stressed than ramets of T3. Evaluating coral stress through reproduction can reveal more sensitive responses than other biological parameters; within reproductive metrics, PGS can be a better stress indicator than egg size. The presence of turf algae strongly impacted the development of gametes and egg size (e.g., PMG in ramets with FTA removal increased almost twofold in comparison with ramets surrounded by FTA in August), most likely exerting negative chronic effects in the long run due to the ubiquity and permanence of turf algae in the Caribbean. These algae can be considered a stressor that affects coral sexual reproduction. Although the effects of turf algae on O. annularis are apparently less severe than those of other stressors, the future of this species is uncertain because of the combined impacts of these effects, the decline of O. annularis populations and the almost complete lack of recruitment.

Reclutamiento del coral Acropora palmata sobre sustratos de dos materiales

espanolDurante el reclutamiento de los corales, las caracteristicas de los sustratos influyen en la densidad y la distribucion espacial de los reclutas. Se evaluo la influencia del material (conchuela y arena) y de 4 combinaciones denominadas grupos (conchuela/arriba, conchuela/abajo, arena/arriba y arena/abajo) en la distribucion de los reclutas del coral Acropora palmata en las 4 areas de los sustratos utilizados (cara superior, caras laterales, caras inferiores y base). En los materiales utilizados, el numero de reclutas por sustrato fue mayor en la conchuela (mediana= 15,0) que en la arena (2,0). En las areas del sustrato, la cara superior en la conchuela y las caras superior y laterales en la arena tuvieron mayor numero de reclutas que las otras areas del sustrato; cabe senalar que las caras superiores fueron las que recibieron mas luz durante el acondicionamiento de los sustratos. En los grupos, el numero de reclutas por sustrato fue mayor en los grupos de conchuela (conchuela/arriba ≈ conchuela/abajo) que en los de arena (arena/arriba > arena/abajo). El relativo bajo numero de reclutas de A. palmata que se obtuvo (12500 m-2) fue, sin embargo, 3-4 ordenes de magnitud mayor que la obtenida en arrecifes del Caribe (3-39 m-2), con una abundancia dominada por corales oportunistas que no proveen las cualidades ecologicas de A. palmata. La siembra en el campo de reclutas obtenidos en acuario permanece como una alternativa viable para recuperar las poblaciones de esta especie en peligro critico de extincion, la cual contribuye sustancialmente en la complejidad estructural de arrecifes coralinos del Caribe. EnglishDuring recruitment of corals, substrate characteristics influence the density and the spatial distribution of recruits. The influence of the material (limestone and sand) and of 4 groups arrangements (limestone/up, limestone/down, sand/up and sand/down) in the distribution of the Acropora palmata recruits in 4 areas of the substrate used (upper face, side faces, bottom faces and base) was evaluated. The number of recruits per substrate was higher in limestone (median= 15.0) than in sand (2.0). Regarding the areas of recruitment, the upper face in limestone and the upper and side faces in sand presented greater number of recruits than the other areas of the substrate; notably the upper faces received more light during the conditioning of the substrates. Concerning the groups, the number of recruits per substrate was higher in limestone groups (limestone/up ≈ limestone/down) than in sand groups (sand/up > sand/down). The relatively low number of A. palmata recruits obtained (12500 m-2) was, however, 3-4 orders of magnitude higher than the recruits obtained in Caribbean coral reefs (3-39 m-2), with an abundance dominated by opportunistic corals that do not provide the ecological qualities fulfilled by A. palmata. The implantation in the field of recruits obtained in aquarium remains as a viable alternative to recover populations of this critically endangered coral species, which substantially contributes to the structural complexity of Caribbean coral reefs.