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Diapause is a reversible developmental arrest faced by many organisms in harsh environments. Annual killifish present this mechanism in three possible stages of development. Killifish are freshwater teleosts from Africa and America that live in ephemeral ponds, which dry up in the dry season. The juvenile and adult populations die, and the embryos remain buried in the bottom mud until the next rainy season. Thus, species survival is entirely embryo-dependent, and they are perhaps the most remarkable extremophile organisms among vertebrates. The aim of the present study was to gather information about embryonic diapauses with the use of a "shotgun" proteomics approach in diapause III and prehatching Austrolebias charrua embryos. Our results provide insight into the molecular mechanisms of diapause III. Data are available via ProteomeXchange with identifier PXD025196. We detected a diapause-dependent change in a large group of proteins involved in different functions, such as metabolic pathways and stress tolerance, as well as proteins related to DNA repair and epigenetic modifications. Furthermore, we observed a diapause-associated switch in cytoskeletal proteins. This first glance into global protein expression differences between prehatching and diapause III could provide clues regarding the induction/maintenance of this developmental arrest in A. charrua embryos. There appears to be no single mechanism underlying diapause and the present data expand our knowledge of the molecular basis of diapause regulation. This information will be useful for future comparative approaches among different diapauses in annual killifish and/or other organisms that experience developmental arrest.

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In Uruguay, a country with a small population, and hence a small scientific community, there were no classical embryologists as such in the past. However, in the decade of the 1950s, a cumulus of favorable conditions gave rise to highly active and modern research groups in the fields of cytology and physiology, which eventually contributed to developmental biology. The advent of a long dictatorship between the 1970's and 1980's caused two things: a strong lag in local research and the migration of young investigators who learned abroad new disciplines and technologies. The coming back to democracy allowed for the return of some, now as solid researchers, and together with those who stayed, built a previously inexistent postgraduate training program and a globally-integrated academy that fostered diversity of research disciplines, including developmental biology. In this paper, we highlight the key contributions of pioneer researchers and the significant role played by academic and funding national institutions in the growth and consolidation of developmental biology in our country.

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BACKGROUND: Many organisms are able to survive in extreme environments by entering a state of dormancy. In dormancy, vital activities are reduced until environmental conditions are compatible with active life. Annual killifishes show a special developmental pattern characterized by a phase of dispersion-reaggregation of the blastomeres that separates epiboly from organogenesis, and the capability to enter dormancy in diapause. High tolerance to environmental stress confers annual killifish embryos the condition of extremophiles. At present, the questions of our research group are focused on the understanding of the mechanisms involved in diapause regulation through an interdisciplinary approach. As a first step, it is necessary to characterize diapauses at morphological and physiological levels and to evaluate induction cues under laboratory conditions. In this context, we characterized diapause I in two Austrolebias species. RESULTS: Our experimental approach to induce diapause I was successful and revealed the co-existence of two diapause I phenotypes named A and B instead of one. These phenotypes showed a tendency for lower total extractable RNA content compared with active developmental stages (80-100% epiboly and early reaggregate). CONCLUSIONS: These phenotypes are alternative diapause I stages and may have ecological relevance because both were found in embryos in natural ponds. Developmental Dynamics 246:848-857, 2017. © 2017 Wiley Periodicals, Inc.

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Over the past 50 years, annual killifishes arose as alternative model organisms for studies of vertebrate biology. The annual fish offers exceptional advantages for studies of genetics, genomics, developmental biology, population dynamics, ecology, biogeography, and evolution. They inhabit extremely variable freshwater environments in Africa and South America, have a short lifespan and a set of unique and fascinating developmental characteristics. Embryos survive within the dry substrate during the dry season, whereas the adult population dies. Thus, the survival of the populations is entirely dependent on the buried embryos that hatch the next rainy season. Although Old and New World species share similarities in their life cycle, they also have different adaptive responses associated with climate-related selective pressures. Therefore, contrasting different species from these areas is essential to understand unique adaptations to heterogeneous environment. A network of laboratories (United States, Czech Republic, Italy, Brazil, Chile, and Uruguay) is working and collaborating on many aspects of the biology of annual fishes. Participating researchers share projects and cross-training undergraduate and graduate students. These efforts resulted in two International Symposia (2010 and 2015) that took place in Montevideo and an international book. Herein, we summarize the progress made by this global community of scientists. Developmental Dynamics 246:807-811, 2017. © 2017 Wiley Periodicals, Inc.

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Evolution of sex determination and differentiation in fishes involves a broad range of sex strategies (hermaphroditism, gonochorism, unisexuality, environmental and genetic sex determination). Annual fishes inhabit temporary ponds that dry out during the dry season when adults die. The embryos exhibit an atypical developmental pattern and remain buried in the bottom mud until the next rainy season. To elucidate genomic factors involved in the sex determination in annual fish, we explored the presence of a candidate sex-specific gene related to the cascade network in Austrolebias charrua. All phylogenetic analyses showed a high posterior probability of occurrence for a clade integrated by nuclear sequences (aprox. 900 bp) from both adults (male and female), with partial cDNA fragments of A. charrua from juveniles (male) and the dsx D. melanogaster gene. The expressed fragment was detected from blastula to adulthood stages showing a sexually dimorphic expression pattern. The isolated cDNA sequence is clearly related to dsx D. melanogaster gene and might be located near the top of the sex determination cascade in this species.

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Annual fish are freshwater teleosts found in South America and Africa that are exposed to an extremely variable environment. They develop and reproduce in seasonal ponds that dry during the summer eliminating the entire adult population. Remarkably, desiccation-resistant embryos survive in these dry ponds that hatch during the next rainy season when the ponds are recreated. Among vertebrates, they represent one of the most remarkable extremophiles. They share several features with other fish models; however, they exhibit unique traits related to their peculiar life cycle. Epiboly is temporally and spatially uncoupled from organogenesis, and the embryos can undergo reversible developmental arrests (diapauses). These attributes make them a useful model to study diverse topics in developmental biology using a comparative and evolutionary approach. In this article, different aspects related to annual fish biology, taxonomy and phylogenetic considerations, reproductive strategy, and developmental characteristics with special focus on arrests, are summarized. The current challenge is to document and determine the factors that generate such high diversity and unique adaptations of annual fish. To understand this complexity, interdisciplinary approaches are being employed taking into consideration evolutionary biology, ethology, reproductive strategies, regulation of developmental mechanisms, and senescence.

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The current knowledge about teleost fish egg envelope is summarized. The paper analyzes the organization and deposition process of the protein composition and genes involved in the synthesis of teleost fish egg envelopes and their role in gamete interaction during fertilization. Pelagic and demersal species that our research group is working with are especially considered. The vertebrate ZP family of proteins, the evolution and relationship among the different genes and their expression are taken into account. We consider fish envelope as a possible biomonitor for ecological contaminants. The biotechnological applications for aquaculture and genomic and post-genomic approaches are auspicious.

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The current knowledge about teleost fish egg envelope is summarized. The paper analyzes the organization and deposition process of the protein composition and genes involved in the synthesis of teleost fish egg envelopes and their role in gamete interaction during fertilization. Pelagic and demersal species that our research group is working with are especially considered. The vertebrate ZP family of proteins, the evolution and relationship among the different genes and their expression are taken into account. We consider fish envelope as a possible biomonitor for ecological contaminants. The biotechnological applications for aquaculture and genomic and post-genomic approaches are auspicious.

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In contrast to most of the previous studies in gymnotiform reproduction, which have been conducted in the tropical region, this study examines a gymnotid from the temperate region in both the natural habitat and the laboratory. The gonadal histology of Brachyhypopomus pinnicaudatus is described for the first time. The male had a paired, lobular testis of the unrestricted spermatogonial type, and females a paired saccular cystovary. Analysis of gonads and their annual cycle enabled us to confirm the breeding season and to conclude that this species is a multiple spawner. Water temperature and photoperiod showed the expected annual cycles for the region. High temperature and a 14 h:10 h L:D photoperiod in the natural habitat coincided with (1) mature gonadal stages, (2) electrophysiological sexual dimorphism: males present a lengthened negative phase in their electric organ discharge (EOD) and (3) decreased temperature sensitivity of the EOD: the waveform does not change when temperature increases above 20 degrees C. Acclimation to sustained high temperature (30 days, 28 degrees C, 12 h:12 h L:D, low conductivity) induced gonad maturation along with EOD dimorphism. Our data show that high environmental temperature is enough to trigger sexual maturity in Brachyhypopomus pinnicaudatus from a temperate climate.

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Realizou-se neste trabalho um estudo histológico do ovário de Macrodon ancylodon, com especial atençäo à ovogênese, aos folículos pós-ovulatórios e à atresia. Na seqüência da maturaçäo ovocitária se evidenciou a existência de 6 tipos celulares: ovogônias, ovócitos basófilos, ovócitos com vitelogênese lipídica, ovócitos com vitelogênese protéica, ovócitos com vitelogênese total e ovócitos hidratados. Os folículos pós-ovulatórios se caracterizaram por estruturas em degeneraçäo. Säo descritos dois tipos de atresia: a hipertrófica e a näo hipertrófica, que aparentemente näo teriam ligaçöes com um possível rol endócrino. As membranas ovulares (teca e granulosa) apresentaram um só tipo celular, sendo negativas as reaçöes histoquímicas para lipídios e colesterol

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