Serotonin and some selected substances known to interfere with its formation (diethyldithiocarbamate) and function (Catron, 5-methyltryptamine, promethazine) were tested for their ability to affect chick embryo morphogenesis during the first 48 h of development. To detect possible differences in sensitivity between the successive morphogenetic events taking place during this period, the treatment was begun at successively more advanced stages corresponding to embryo ages of between 4 and 30 h incubation. In all cases, the treatment was terminated at an embryo age of 48 h incubation. The treatment was performed both in ovo and in vitro.With some exceptions, the substances induced malformations of the same characteristic types. The developmental processes subjected to disturbances included blastoderm expansion, primitive streak formation, neurulation with brain formation, and somitogenesis. At the cellular level, the malformations can be traced to delayed yolk degradation, impaired formation and function of microvilli, and impaired ability of the embryo cells to change shape.All of the tested chemicals can be expected to interfere with intracellular levels of serotonin. They obviously interfered with decomposition of the yolk granules, recognized centres for intracellular serotonin formation and we therefore conclude that the observed morphogenetical disturbances are ultimately due to impairment of the endogenous serotonin formation. We suggest that, in morphogenesis, serotonin primarily promotes the activity of microtubules and microfilaments.
Serotonin distribution in early Ophryotrocha embryos was investigated with fluorescence microscopy based on formaldehyde gas treatment of the embryos, and with light- and electron-microscopic autoradiography after the embryos had been treated withH-5-hydroxytryptophan.Sections of early cleavage embryos showed serotonin-specific fluorescence all over the blastomeres, but it was mainly concentrated on yolk granules, and to a lesser degree on lipid drops and vacuoles. In 2-8 cell embryos, marked regional concentration of serotonin fluorescence was noticeable along the completed cleavage furrows.The autoradiographs confirmed the picture of the yolk granules as the principal site of serotonin formation and serotonin accumulation; considerable amounts were also associated with their decomposition products, i.e. lipid drops, vacuoles, and vesicles, whereas major cell organelles, e.g. mitochondria, were almost totally lacking. Of cytoplasmic structures in the blastomeres without apparent yolk granule origin, only microfilaments, particularly those amassed along the cleavage furrow, showed consistent and significant association with formed serotonin. This suggests a connexion between serotonin and microfilaments and might imply that in early embryo cells the fundamental contractile machinery is controlled by serotonin gradually released from the yolk granules.Within the blastomere nuclei, moderate amounts of serotonin were demonstrated with both fluorescence microscopy and autoradiography.The monoamine oxidase (MAO) inhibitor catron (phenylisopropylhydrazine), used to intensify the autoradiographic picture of serotonin in the Ophryotrocha embryos, markedly increased intragranular serotonin accumulation, but also retarded yolk granule disintegration and delayed the cell cleavage process. In embryos barely able to cleave after treatment with catron, ultrastructural analysis demonstrated that membrane formation at cell cleavage depends on influx of material from the nearby disintegrating yolk granules.
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