Browsing by Person "Tisler, Matthias"

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Funktionelle Analyse der Histondeacetylase 6 sowie experimentelle Modellierung von Lateralitätsdefekten während der Links-Rechts-Achsenentwicklung von Xenopus laevis und Paracentrotus lividus
(2017) Tisler, Matthias; Blum, Martin
Vertebrates display an asymmetric positioning of the visceral organs, which is also denominated as left-right body axis. During embryogenesis, an asymmetric gene expression is detectable that is initiated by an evolutionary conserved mechanism of symmetry breakage, which is conserved among deuterostomes. During neurula stages, rotating motile mono-cilia at the so called left-right organizer (LRO) generate an asymmetric stimulus known as extracellular leftward fluid flow that is essential for the unilateral left asymmetric gene expression of the Nodal cascade. Spontaneous mutations or the experimentally induced loss of function of genes influencing ciliogenesis at the LRO, the induction of the Nodal cascade or its propagation lead to left-right defects. Left-right defects are frequently observed in human conjoined twins. Thoracopagous, dicephalic conjoined twins display defects in the arrangement of the inner organs, that are solely reported from the twin located to the right side. While left twins orient the inner organs wildtypically, right twins show a randomization of the left-right axis. The functional cause of the inverted arrangement regarding the right twin has remained enigmatic. It has been hypothesized that the observed laterality determination in conjoined twins, like in wildtype embryos, was dependent on leftward flow. In the course of this thesis, the known unilaterlal left-sided induction of the Nodal cascade in the left conjoined twin, as in singelton embryos, can be linked to leftward flow. The artificial induction of a second body axis leads to a subsequent duplication of the LRO during development. During flow stages endogenous and induced LROs locate in close proximity and display a partial fusion of cell populations. Anti-sense Morpholino Oligomeres or methylcelluose mediated loss of cilia motility lead to a loss of markergene expression in the left-lateral plate mesoderm of the left twin. By combining differential gain- and loss-of-function strategies, it was possible to link the establishment of laterality in conjoined twins to the leftward flow and, moreover, to manipulate it an a predictable manner. The cause of this hitherto enigmatic laterality defects in conjoined twins can therefore be explained by the evolutionary conserved mechanism of left-right establishment. Although the general mechanism of symmetry breakage has been characterized, novel candidate genes are continously beeing identified that act at a specific sequence of this process. The candidate gene histonedeacetylase 6 (hdac6) was shown to impact on left-right development. Anti-sense Morpholino Oligomere induced loss-of-function experiments led to left-right defects in a dose dependent manner regarding, the induction of the genes of the Nodal cascade, indicating a function of hdac6 before fluid flow induced regulation of dand5 mRNA. Taken together: histonedeacetylase 6 acts as modulator of canonical Wnt-signaling in the transcriptional induction of the Wnt-dependent transcription of foxj1, a master control gene of the biogenesis of motile cilia. Loss of Hdac6 leads to defects regarding the ciliogenesis of motile cilia at the LRO as well as the multiciliated epidermis of the embryo. The here presented results represent the first developmental hdac6 loss-of-function phenotype, which was so far not know from Hdac6-/- mice. These experiments shed a new light on the differential in vivo function of this unique histondeacetylase during development. Even though the asymmetric positioning of the inner organs is restricted to vertebrates, the asymmetric expression of the Nodal cascade turns out to be evolutionary conserved among deuterostomes. Comparable to vertebrate species, larvae of the sea urchin (Paracentrotus lividus, Echinodermata) display an asymmetric expression of the Nodal cascade in the ectoderm an during gastrula stages. Experiments from this work could demonstrate that also in sea urchin embryos the asymmetric gene expression depends on motile cilia. The archenteron of gastrula stage embryos was identified and described as homologous structure to vertebrate LROs. Deciliation experiments at different time points of development induce laterality defects and point towards a symmetry breakage during early gastrulation. By this experiments, the cilia dependent establishment of left-right asymmetry is described as a common synapomorphy of the deuterostomes beeing conserved from sea urchin to vertebrates, shedding a new light on the establishment of asymmetric gene expression.
MMP21 behaves as a fluid flow transported morphogen to impart laterality during development
(2025) Ott, Tim; Brugger, Amelie; Szenker-Ravi, Emmanuelle; Kurrle, Yvonne; Aberle, Olivia; Tisler, Matthias; Blum, Martin; Whalen, Sandra; Bouvagnet, Patrice; Reversade, Bruno; Schweickert, Axel
Heterotaxy (HTX) is frequently caused by deleterious variants in the gene encoding Matrix metallopeptidase 21 (MMP21). However, the underlying pathomechanism has not been ascertained. In this study, we report on a novel HTX-associated MMP21 knockout allele in humans and investigate the peptidase’s role during laterality development using Xenopus embryos as animal model. The targeted inactivation of mmp21 in f0 mutant Xenopus successfully phenocopied the human HTX condition, yet the cilia-driven leftward fluid flow, which initiates asymmetric gene activity at the left-right organizer (LRO), was unaltered in mmp21 null frogs. Instead, our analysis of downstream events revealed that flow response, the left-sided repression of dand5, could not take place. Remarkably, gain-of-function experiments demonstrated that Mmp21 spreads over LRO cells and triggers flow response. Additionally, Mmp21 functions upstream of Cirop, another metallopeptidase, which we found specifically localized to LRO cilia. Thus, our findings suggest that Mmp21 may be the long-sought morphogen, which is actively transported by the leftward fluid flow to Cirop-laden cilia, in order to specify the left side of the embryo.