Confocal-laser-scanning-microscope

• Subject Area: Neurosciences
• Term: Funded in 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 499148687

The human body is composed of many different cell types which communicate with each other. In particular, the brain consists of billions of neurons and non-neuronal cells which are interconnected and require tight and precise regulation of cellular processes. RNA editing is a cellular process that diversifies gene function by enzymatic deamination of cytidine or adenine. This can result in changes of protein structure and function. Altered RNA editing is becoming increasingly associated with all kind of disease, but most approaches use advanced sequencing technologies to analyze bulk material. However, it is also becoming progressively evident that changes in RNA editing have to be analyzed and considered in a cell type specific way. Excitatory and inhibitory transmission between neurons in the brain needs to be thoroughly regulated and coordinated. Deregulation of this coordination ultimately results in nervous system disorders. A core aspect of our work concerns the study of the brain at the molecular level, by investigating C-to-U RNA editing and alternative RNA splicing. In particular, we analyze key components of the molecular machine responsible for inhibition of electrical impulses, that are glycine receptors (GlyR) and GABA type A receptors (GABA(A)R) as well as the postsynaptic scaffold protein gephyrin. We elucidate the function of these molecules in physiological and pathophysiological processes on a molecular, cellular and systemic level. Temporal lobe epilepsy (TLE) is a devastating neurodegenerative disease that severely deteriorates life quality due to unpredictable occurrence of seizures and resulting cognitive dysfunction. Moreover, epilepsy patients suffer from severe psychiatric comorbidities including anxiety and depression. Most epilepsy syndromes have no discernable genetic component. This indicates that epileptogenesis is governed by disease-promoting molecular and cellular mechanisms of neuronal plasticity, which may vary from patient to patient and involve cell type-specific mechanisms, resulting in diverse clinical pictures of cryptogenic/idiopathic TLE. Recently, we developed in our lab molecular tools and used advanced chemo-biological approaches to visualize and characterize C-to-U RNA editing in general, and regarding GlyRs in particular. We propose to identify critical cell types with TLE-dependent alterations in RNA editing at the single cell level in vitro and ex vivo. To take this project to the next level we are also developing a novel molecular tool for the detection of C-to-U RNA editing in a cell compartment-specific way down to the level of single synapses. Altogether, our recently developed advanced molecular tools should allow us to gain knowledge about cell type-specific pathophysiological mechanisms in TLE and provide starting points for novel therapies.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Konfokales-Laser-Scanning-Mikroskop
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Technische Universität Braunschweig
• Leader: Professor Dr. Jochen Meier