Ongoing studies

Professor Per Borghammer and ph.d.-student Mie Just Pedersen, Aarhus University, Department of Clinical Medicine - PACE - Lundbeck Foundation Parkinson's Disease Research Center

PACE is a center dedicated to multi-disciplinary study of Parkinson’s disease and the related disorder, Lewy body dementia. During the past five years, we have pioneered the use of the Danish Brain Collection for the study of neurodegenerative disorders. We have already identified 140 Lewy body-positive cases, the defining feature of Parkinson’s disease. We have validated advanced antigen-retrieval protocols, which allows high quality immunohistochemical staining for Lewy pathology and Alzheimer-type protein pathologies. The quality of our stains are now on par with that from modern brain banks, clearly demonstrating the utility of the brain bank despite the extended storage period. We have planned a range of studies in Parkinson’s disease and Alzheimer’s disease in the coming years, which would greatly benefit from an optimized digital and research infrastructure.

Professor Jens Randel Nyengaard, Aarhus University, Department of Clinical Medicine, Core Centre for Molecular Morphology, Section for Stereology and Microscopy

We are working with a well-characterized collection of post-mortem human brain samples from “The Brain Collection” in Odense, Denmark. This unique resource provides an exceptional opportunity to study molecular changes across different stages of Parkinson’s disease.
To visualize and quantify protein markers, we will use: 

• High resolution confocal microscopy and immunohistochemistry to localize Synaptojanin 1, OCRL, and PIKfyve at cellular and subcellular levels 

• These imaging results will be correlated with hallmark Parkinson’s pathology, e.g., α-synuclein fibrils, dopaminergic neuron loss.

Professor Christoph Beier, University of Southern Denmark, Research unit of Neurology (Odense)

The Danish Brain Collecetion enables us to investigate the consequences and mechanisms of long-standing seizures in an era prior to the widespread use of modern medications. This has led to the identification of tau accumulation as a likely consequence of long-standing seizures and a possible contributor to cognitive decline. Future studies will focus on the mechanisms of seizure-induced neuronal damage and degeneration.

Associate Professor Christina Kjær, University College Copenhagen, Faculty of Health, Department of Technology

Our understanding of disorders and diseases of the brain comprising Alzheimer’s, epilepsy, major depression, and schizophrenia has been revolutionized by the past decades rapid development of Next-Generation-Sequencing (NGS) technologies. There is still a high unmet need to identify new drug targets for the development of drugs with higher efficacy and fewer side effects. For this, there is an enormous untapped potential in ‘the Danish Brain Collection’, on which we via methods and protocols developed for evolutionary studies, recently – and for the first time – successfully extracted DNA and RNA from formalin fixated paraffin embedded samples from the Danish brain collection in amounts and qualities applicable for NGS analysis.

Primary investigator: Mikkel Vestergaard Olesen

The purpose of the research project is to make use of stereological methods as well as molecular biological techniques to investigate disease relevant brain regions, including central gray structures, in post-mortem brains of schizophrenic patients and control subjects. We will include brains from untreated and treated schizophrenic patients, in order to determine possible effects of medication.
The project has been approved by the Regional Ethics Committee of the Capital Region of Denmark.

Estimation of the total number of cells (neurons, oligodendrocytes, astrocytes and microglia) in the basal ganglia of the brain using stereological counting methods.

We have identified and obtained 13 brains as well as associated autopsy reports from non-medicated schizophrenic patients (died before 1952) in the brain collection at SDU. In addition, we have identified and prepared 12 brains from age- and gender-matched controls from our own brain bank at Bispebjerg Hospital. As the autopsy report from the schizophrenic patients does not contain detailed clinical information (e.g. any other medical treatment or diseases that may affect the CNS), the relevant medical records were obtained from the National Archives (Viborg, Odense and Copenhagen), scanned and analyzed.

We have also:

• Performed a detailed visual inspection of the formalin-fixed tissue so that brains without intact central regions (due to previous pathological examinations) are omitted. We are able to use 11 out of 13 schizophrenic brains, corresponding to ~ 85%.
• Sectioned the central brain regions into 40-μm thin consecutive sections. This is done exhaustively and gives approximately 1000 sections per. brain when collecting every sixth section. Until now this is completed in 6 out of 11 schizophrenic brains, corresponding to ~ 55%.
• Giemsa-stained a fraction of the sections in each brain to verify the usability for volume estimation and cell identification. In 4 out of 6 schizophrenic brains we are able to identify the neurons (corresponding to 67%) and in 2 out of 6 brains there are identifiable glial cells (corresponding to 33%).

Based on the numbers above, we will thus be able to quantify the neurons in ~ 57% and the glial cells in ~ 28% of the brains from schizophrenic patients without medical treatment (formalin fixed >70 years).

Conclusions

We are able to complete the current project, but not within the specified time frame of the application. This is partly due to the characteristic of the tissue and the susceptibility to staining, which means that only a relatively small number of the obtained brains can be used. A stereological estimation of the neurons is possible, whereas counting of the glial cells will be subjected to particularly great uncertainty.