Studies of In Vitro Embryo Culture of Guppy (Poecilia reticulata)
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Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggswithin the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk,before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos withresources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, havebeen limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females wereincubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). Weinvestigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether theywould hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower invitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread indevelopmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although inculture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire periodof their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems welldeveloped, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in thesetwo culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appearsto be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.
Abstract
Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggswithin the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk,before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos withresources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, havebeen limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females wereincubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). Weinvestigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether theywould hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower invitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread indevelopmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although inculture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire periodof their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems welldeveloped, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in thesetwo culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appearsto be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.
참고문헌 (15)
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