RESUMEN

In vertebrates, only few experiments have been performed in microgravity to study the embryonic development from fertilization. To date, these concern only amphibian and fish. We report here a study on the embryonic development of Pleurodeles waltl (urodele amphibian) eggs oviposited in microgravity. The experiment was performed twice on board the Mir space station and the data obtained included video recording and morphological, histological and immunocytological analyses. The data confirm that the microgravity conditions have effects during the embryonic period, particularly during cleavage and neurulation, inducing irregular segmentation and abnormal closure of the neural tube. Moreover, we observed several abnormalities hither to undescribed corresponding to cortical cytoplasm movements, a decrease of cell adhesion and a loss of cells. These abnormalities were temporary and subsequently reversible. The young larvae that hatched during the flight displayed normal morphology and swimming behavior after landing. The results obtained in the urodele Pleurodeles waltl are in accordance with those observed earlier in the anuran Xenopus laevis and in the fish Oryzias latipes.

Asunto(s)

Embrión no Mamífero/fisiología , Sistema Nervioso/embriología , Pleurodeles/fisiología , Vuelo Espacial , Ingravidez , Animales , Fenómenos Fisiológicos Celulares , Colina O-Acetiltransferasa/metabolismo , Fase de Segmentación del Huevo , Embrión no Mamífero/anomalías , Desarrollo Embrionario , Femenino , Larva , Actividad Motora , Óvulo , Pleurodeles/anomalías , Pleurodeles/embriología , Pleurodeles/crecimiento & desarrollo , Natación

RESUMEN

The nucleocytoplasmic hybrid combination performed by nuclear graft between B. bufo nucleus and B. calamita egg is 95% lethal during gastrulation mostly at the young gastrula stage (75%) and to a lesser extent at the yolk plug stage (20%). The heterologous B. calamita cytoplasm was treated or fractioned by different techniques including centrifugation, nucleic acid extraction, ammonium sulfate treatment and heating. The treated cytoplasmic fractions were injected in B. bufo fertilized eggs in order to check their influence on morphogenetic capacities of the B. bufo nucleus. Injection of hyaline cytoplasmic fraction obtained from B. calamita centrifuged eggs blocks the B. Bufo fertilized egg gastrulation in similar rates as in nucleocytoplasmic association performed by interspecific nuclear grafting. Gastrulation, however, is not affected by injection of total nucleic acids extracted from this hyaline fraction. The soluble cytoplasmic fraction obtained after 60% ammonium sulfate precipitation blocks the development at the beginning of gastrulation, this property being inactivated by heating for 5 min at 60°C. Supernatant from 70% ammonium sulfate precipitation blocks the development at the yolk plug stage and is inactivated by heating for 5 min at 80°C. These experiments suggest that two distinct factors in B. calamita eggs are involved in developmental arrest of injected B. bufo fertilized eggs, these two factors respectively acting on the beginning and on the end of gastrulation.

RESUMEN

Nucleocytoplasmic interactions have been studied through nuclear and cytoplasmic grafts. Bufo bufo nuclei transplanted in Bufo calamita eggs provoked 95% lethality during gastrulation. Influence of the cytoplasm in such a nucleocytoplasmic incompatibility has been studied through cytoplasmic fractionation and grafting of the fractions obtained. Fractionation of egg cytoplasm gave three fundamental fractions: lipid, hyaline cytoplasm and vitellus. After injection of B. calamita fractions into B. bufo fertilized eggs, the hyaline cytoplasmic fraction provoked a gastrulation block in the same ratio (95%) as in the interspecific nuclear graft experiment. Influence of the hyaline cytoplasmic fractions of both species has been tested on nucleocytoplasmic and natural hybrid developments. The B. calamita hyaline cytoplasmic fraction provoked in each case a developmental block at the beginning of gastrulation. The B. bufo hyaline cytoplasmic fraction acts later at the yolk plug stage and in some cases partially corrects the developmental bolck or exogastrulation of young hybrid gastrula. It can be generalized that in both kinds of hybrids, the hyaline cytoplasmic fraction of B. calamita exerted a lethal effect on B. bufo nucleus.

RESUMEN

Cytological evolution and DNA synthesis activity of the female pronucleus were analyzed in experimentally activated virgin eggs of Pleurodeles. Pricking with a micropipette is a positive treatment for activation comparable to electric shock. As in normally fertilized eggs, the evolution of a female pronucleus is related to normal DNA synthesis. Thus, the fact that the female gametic nucleus does not achieve the first mitotic cycle, at the end of migration, cannot be related to initial failure of DNA synthesis. Normal mitoses of this nucleus are obtained in an activated egg in which development is insured by grafting a somatic nucleus. These results are discussed in terms of activation factors.