Gene and protein regulation i mouse oocytes and preimplanation embryos.

We study early development in mammals. We are especially interested in oocyte maturation, fertilization and the first embryonic cell division. This relative short period in development is referred to as the maternal-zygotic transition (MZT). An important shift in control of gene regulation takes place during this period, where the control is gradually transferred from maternal to zygotic. Our main focus is the establishment of heterochromatin and associated epigenetic modifications. We use a combination of molecular and biochemical approaches as well as more advanced micromanipulations of embryonic cells. The vision with our research is to identify genes that play essential roles in the regulation of the MZT. Mistakes in these early mechanisms give rise to involuntary childlessness, and the research seek to elucidate the regulations in reproduction.

Another part of our research investigates the relationship between P-type ATPase membrane proteins and their roles in neurological diseases. Mutations in ATP1A3 trigger the familial Rapid-Onset of Dystonia Parkinson (RDP), whereas mutations in ATP1A2 trigger familial hemiplegic migraine type 2 (FHM2). For this purpose, we are currently generating conditional knock-out mice; a design that permits a cell specific elimination of ATP1A2 and/or ATP1A3 isoforms.

Redox regulation of protein activity

Reactive oxygen species (ROS) - which are a group of reactive oxygen-containing compounds – are believed to play an important part in the ageing process and in a number of disease states, including rheumatoid arthritis, atherosclerosis, and neurodegenerative diseases. These compounds have the ability to introduce irreversible chemical modifications in bio-molecules, whereby the normal function of these can be altered or destroyed. However, it is now clear that ROS also play an important role in normal physiology as, e.g., cell propagation and differentiation. Therefore, under normal physiological conditions a delicate balance between antioxidants and ROS (oxidants) exist, prohibiting irreversible and detrimental modification of bio-molecules.

In addition to the introduction of irreversible modifications, ROS are also known to introduce a number of reversible chemical modifications. The focus of the ongoing research is to investigate how these reversible modifications can modulate or regulate protein activity. The overall perspective of the research is to establish how the protein complement in the body responds in diseases where ROS are believed to play an important role in the etiology. The research conducted use a number of methodologies in protein chemistry and molecular biology as well as cell culture systems.

Proteomics to study disease associated protein perturbations - Discovery and characterisation of biomarkers and drug targets

The proteomics research group lead by Prof. DrMedSc Bent Honoré has as main focus the identification, characterisation and functional analyses of differentially expressed proteins in a set of diseases. Such proteins may be biomarkers for the disease or suitable targets for drug treatment. The research is conducted along two lines using studies on pathological patient tissue as well as basic laboratory research. The tissue samples are collected and analysed in collaboration with a number of clinical departments covering especially cardiovascular diseases, eye diseases and cancer. In addition we perform basic scientific studies involving laboratory animals and cell culture models as well as molecular studies on selected proteins.

Techniques: A variety of techniques are used with the major ones being 1) two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), an electrophoretic technique to separate proteins with high resolution based on isoelectric point and molecular mass and 2) liquid chromatography - tandem mass spectrometry (LC-MS/MS) for protein identification based on analysis of the sequence of amino acids of smaller peptides. In addition, a vide variety of techniques are used for basic studies including biochemistry, molecular biology and cell biology.

Diseases: The disease studies include 1) cardiovascular diseases (heart failure, aortic aneurysms and diabetic arterial disease), 2) eye diseases (melanoma, retinoblastoma, corneal-, vitreal- and retinal diseases) (Henrik Vorum and Nakul Mandal) and 3) cancer including lymphomas and carcinoma of the colon (Maja Ludvigsen).

Basic analyses on putative biomarkers and drug targets: A number of the identified putative biomarkers and drug targets discovered using the proteomic techniques are further analysed and characterized. Recent studies concern the CREC protein family of EF-hand calcium binding proteins, mainly localised in the secretory pathway that include reticulocalbin (Gry Hansen), ERC-55 (Maja Ludvigsen) and calumenin (Gry Hansen). These studies are conducted to functionally characterise putative biomarkers further by revealing subcellular localisation, identifying protein binding partners, etc.