25–30 May 2020, Spetses Island, Greece
Phone: 0043 1 40160 37713
Our research centers on the functional and dynamic architecture of chromatin in cell nuclei during differentiation processes. Regular and aberrant differentiation processes as well as organ development reflect the particular importance of coordinated chromatin organization.
Structural and functional constituents of nuclei determine chromatin organization and are in turn influenced by the dynamic landscape of chromatin. The interaction of chromatin and the different constituents result in an important layer of epigenetic gene expression control, which is characteristic for specific cell types. The interplay between the cytoplasm / nuclear membrane / nuclear lamina / chromatin has been of special interest for us for considerable time. Thus, one of our current projects focuses on the characterization of LEM-domain proteins and their significance for mitotic progression.
Currently, these questions are investigated at cellular level using imaging and molecular methods. Differentiation processes are also studied in developing organisms (chicken, transgenic fish).
Olympus FV3000 upright confocal microscope
FEI Tecnai 20 transmission electron microscope (+Amira software for 3D reconstruction)
Nikon Eclipse Ti inverted fluorescence microscope for live cell imaging
Nikon Eclipse800 upright fluorescence microscope
Olympus VS120 automated slide imaging system
complete histology unit (paraffin embedding + preparsation of stained sections)
1) Various detection methods for transcriptional activities: immunodetection (e.g. of epigenetic markers) at light and electron microscopic level, DNA and RNA in situ hybridization at light and electron microscopic level, production of stable transgenic cell lines expressing fluorescently-tagged proteins
2) Production of knock out cells with the CRISPR/Cas9 system
3) Life cell imaging
4) Developmental model systems for precise spatio-temporal manipulations and for genetic manipulations.
In ovo electrophoresis of RNAi constructs or morpholinos will be used in chicken embryos in order to be able to introduce e.g. inhibitors of developmentally relevant factors.
The novel aging model system Nothobranchius furzeri (killifish) is an extremely short-lived organism providing the possibilities for genetic interference and for subsequent studies of possible effects during the entire life span.
We have the expertise in preparing and evaluation of samples for a wide repertoire of microscopic methods, including both light- and electron microscopy.
Basic light and electron microscopy plus specialized techniques (EM-tomography) can be performed in our lab.
Roles of epigenetic factors in higher order chromatin arrangement of differentiating cells
We are studying the interdependence of epigenetic determinants such as histone variants (focus: H3 variants), histone modifications and gene expression in cells undergoing regular and aberrant differentiation.
The interplay of chromatin structure and gene expression in developing organisms
Epigenetic determinants we are studying include histone variants, in particular the H3 subtypes, histone modifications specific for transcriptionally active versus inactive chromatin and histone deacetylase (HDAC) inhibitors class I. These determinants are correlated with chromatin architecture.
The structure/function interplay is studied throughout embryonic development (model system chicken) as well as aging (model system Nothobranchius furzeri).
Tissues/Organ systems we are currently investigating are neural crest derivatives as well as heart and limb bud development.
Nuclear lamina – LEM-domain proteins
Several proteins of the inner nuclear membrane (INM) belong to the LEM-domain (Lap2-Emerin-MAN1) protein family. More specifically, we are investigating the LEM-domain protein ANKLE2 which seems to be involved in the regulation of nuclear envelope formation after mitosis.
Up to now researchers coming from extern do not have to pay a fee for the use of equipment. A collaboration (co-authorship, acknowledgement) and/or a financial contribution to consumables are discussed individually.
25–30 May 2020, Spetses Island, Greece