In vitro techniques – analysing human and rodent brain tissue
Genes carry the fundamental information to build and maintain cells, including neurons. Any gene of a cell can undergo changes in the cells lifetime, i.e. it can become mutated. Whereas many mutations are harmless, some have the potential to severely interfere with the normal function of a neuron, a group of neurons, or the whole brain, forming the underlying basis to develop disease.
To understand the effects of specific genetic mutations, we work with knockout animals that have targeted mutations in genes that are high-potential candidates for disease. Particularly, we are studying the role of a Neuregulin-1 knockout and GPCR12 knockout in health and disease; the genotype of these knockout animals is confirmed using PCR. To gain more insight into genetic mutations in certain areas of the brain and their association with disease development, we measure region specific gene expression using In situ hybridization.
Proteins are essential for every process within a cell as well as cell-cell communication, including neurotransmission. Proteins are essential for developing and maintaining neuronal structure and can function as transmitters, receptors, modulators and mediators in the brain. Therefore, identifying proteins, that are (1) changed in diseased brains, (2) get modulated by existing drugs or (3) are potential targets for modification by new drugs, is the central topic in our research.
To examine proteins, particularly receptor protein, density, expression levels and functional capacity, we use Receptor Autoradiography, G-protein assays, Western blot, Immunohistochemistry, ELISA and RIA assays. Using these techniques, we examine tissue from both patients and various mouse and rat models for certain diseases.
- Cell culture
Cell lines are fundamental to gain detailed information about disease-related pathways on a cellular level. Human cell lines and primary cell culture (e.g. hippocampal cells from mutant mice) are a primary focus in our laboratory to study the role and interactions of disease-related proteins.
Staining of brain slices or cells, previously fixed, is essential to examine neuronal morphology and connectivity. This can be achieved by staining certain cellular elements of interest. Golgi staining is employed to study the morphology and connectivity of brain tissue from our animal models. Cell staining is used to visualise cell morphology and cell components after disease-relevant chemical treatment.
- Human Brain tissue
Applying many of the above techniques to human post-mortem brain tissue from people suffering disease, allows us to understand what processes are disrupted in the diseased brain. This then gives us something to model in our animal models and gives us targets to investigate if novel treatments can in fact reverse any of these disease-related processes.
In vivo techniques – studying animal models
- Animal models
Animal models mimic certain symptoms of human diseases and allow researchers to: (1) gain an understanding of the underlying mechanisms of disease, (2) establish diagnostic criteria/tests for disease, (3) gain an understanding of the underlying mechanisms of disease treatments and (4) develop and trial new treatments. In our lab, we use genetic, pharmacological, metabolic and behavioural rat and mouse models of disease.
Behaviours are all interactions of an individual with its environment. All environmental stimuli are integrated and processed by the brain. All active and reactive behaviour is controlled by the brain and reflects therefore a healthy or pathological mental state. To study disease-related behaviour in our animal models of disease, we use behavioural tests for emotionality, social behaviour, cognition and locomotion.
Invasive Techniques / Surgery
Brain microdialysis is used to determine the factors/neurotransmitters that are released from neurons in the course of neuronal communication. This allows us to follow neurochemical changes in the brain due to certain disease-relevant stimuli or drug treatment. In our lab, we use microdialysis in freely moving animals, which allows the correlation of disease-related behaviours with ongoing neurochemical changes in the brain.
Whereas the preferred method of drug application for an animal is via food or drinking water, some novel and experimental drugs cannot pass the blood-brain barrier. This requires the insertion of an intracranial cannula allowing intracerebral or intraventricular application.