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Evolution of the gene regulatory network underlying the formation of the gastrula organizer
Macháčová, Simona ; Kozmikova, Iryna (advisor) ; Krylov, Vladimír (referee) ; Buchtová, Marcela (referee)
During gastrulation, the vertebrate embryo is organized from the clump of cells into a bilaterally symmetric body. This organization process is driven by the gastrula organizer. Its establishment is induced by maternal Wnt/β-catenin signaling and Nodal/Activin signaling localized in the presumptive dorsal region of the embryo. The regulative environment then triggers the expression of the organizer-specific genes which create morphogen gradients in the embryonic body and therefore give each cell positional information. However, the evolution of vertebrate organizer establishment remains vague. Here we aim to test the compatibility of the invertebrate cis-regulatory modules with the vertebrate gene regulatory network (GRN). We introduced fluorescent reporter genes under the control of the invertebrate regulatory sequence of organizer-specific genes into a vertebrate model to observe their behavior in the context of the vertebrate GRN. We found and functionally verified a 500 bp-long amphioxus sequence (an enhancer) that is necessary and sufficient to drive a correct Chordin gene expression in the gastrula organizer in zebrafish. Chordin is a prominent organizer-specific gene antagonizing Bone Morphogenetic Protein (BMP) signaling. We tested also other invertebrate genes for their compatibility with...
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Comparative development & evolution of adenohypophysis in vertebrates
Brezarová, Dominika ; Černý, Robert (advisor) ; Kozmikova, Iryna (referee)
Adenohypophysis is very important gland in vertebrate head. In general adenohypophysal development is described together with formation of the primary mouth. Adenohypophyseal placode migrates together with stomodeal placode in ventral direction. Stomodeal placode starts to invaginate and from oral ectoderm adenohyphyseal placode invaginates to form Rathke's pouch. Pouch is then separated from mouth and migrates towards the brain to make adult hypohyseal gland. Origin of adenohypophysis is therefore in oral ectoderm, but in some vertebrates species is this development little different and adenohypophysis can even be endodermal. In some vertebrate groups, like amphibians or fishes there's no sign of Rathke's pouch, thus early migrations of adenohypophyseal placode can be different. These differences are there maybe because these groups don't have shallow invagination of stomodeum like other vertebrates. There are some homological structures in other chordate groups, for example Hatshek's pit in amphioxus or neural gland in tunicates, these structures has, in contrast with vertebrates, endodermal origin. Key words: adenohypohysis, placodes, primary mouth, craniofacial development, Rathke's pouch, vertebrates
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Bmp signaling in the evolution of chordate axial patterning
Kozmikova, Iryna ; Kozmík, Zbyněk (advisor) ; Bryja, Vítězslav (referee) ; Markoš, Anton (referee)
Formation of a dorsoventral axis is a key event in the early development of most animal embryos. In vertebrates, early separation of cell fate domains precedes specification of ectoderm to neural and non-neural as well as mesoderm to dorsal and ventral during development. Maintaining such division with the establishment of an exact border between the domains is required for the formation of highly differentiated structures such as neural tube and chorda. In the cephalochordate amphioxus, genes encoding Chordin, Bmps and transcription factors downstream of Bmp signaling such as Vent are expressed in patterns reminiscent of those of their vertebrate orthologues. However, the key question is whether the conservation of expression patterns of network constituents implies conservation of functional network interactions, and if so, how an increased functional complexity can evolve. Here, we therefore investigated the role of Bmp signaling in axial patterning and cell fate determination in amphioxus, the basal chordate possessing a centralized nervous system and dorsal mesoderm. Using heterologous systems, namely by reporter gene assays in mammalian cell lines and by transgenesis in medaka fish, we have compared the gene regulatory network implicated in dorsoventral patterning of the basal chordate...
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