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).
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. Advanced light microscopy (superresolution microscopy) can be carried out at certain facilities of other consortium partner institutions. For example, collaboration with the Institute of Molecular Genetics (Prague) is well established.
Superresolution with option for life cell imaging
Light-sheet microscopy for embryonic specimen
EM Array tomography
Advanced image processing
Murko, C., Lagger, S., Steiner, M., Seiser, C., Schoefer, C. and Pusch, O. (2013). Histone deacetylase inhibitor Trichostatin A induces neural tube defects and promotes neural crest specification in the chicken neural tube. Differentiation; research in biological diversity 85, 55-66.
Philimonenko, A. A., Janacek, J., Snyers, L., Almeder, M., Berger, W., Schmidt, W., Schofer, C., Hozak, P. and Weipoltshammer, K. (2011). Chromosomal dynamics of cell cycle regulator gene p21 during transcriptional activation. J Struct Biol 173, 382-90.
Snyers, L., Vlcek, S., Dechat, T., Skegro, D., Korbei, B., Gajewski, A., Mayans, O., Schofer, C. and Foisner, R. (2007). Lamina-associated polypeptide 2-alpha forms homo-trimers via its C terminus, and oligomerization is unaffected by a disease-causing mutation. J Biol Chem 282, 6308-15.
Snyers, L., Zupkovitz, G., Almeder, M., Fliesser, M., Stoisser, A., Weipoltshammer, K. and Schofer, C. (2014). Distinct chromatin signature of histone H3 variant H3.3 in human cells. Nucleus 5.
Weipoltshammer, K. and Schofer, C. (2016). Morphology of nuclear transcription. Histochemistry and cell biology 145, 343-358.