Imaging/High content screening system

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 520854718

We apply for a microtiter plate-based brightfield and fluorescence imaging/high content screening system. This system allows for fast, partially automated imaging, quantification and analysis of cellular characteristics as well as small organisms such as the nematode Caenorhabditis elegans. Our group studies the relevance of Adhesion G protein-coupled receptors (aGPCRs) to understand how they translate signals into function in distinct biological processes. Adhesion GPCRs are an intriguing class of GPCR with diverse roles in health and disease. Despite their obvious (patho-)physiological importance, the details on their activation, signals and the physiological consequences thereof are still only sparsely understood. Our work analyses the molecular mechanisms of these unique GPCRs and aims to shed light on the question how they integrate signals in physiological processes. To understand how the receptors control physiological functions, we are elucidating the roles of aGPCRs in different contexts: on a cellular level, in tissues and organs, and in entire organisms such as the nematode Caenorhabditis elegans. Thereby, our analyses strongly depend on structure-function analyses of aGPCRs. For this purpose, we first investigate the effects the absence of an aGPCR has on cells and on the nematode C. elegans. This is mainly achieved by phenotyping assays. Subsequently, the effects of different modified or naturally occurring receptor variants on these phenotypes are investigated. In a second approach, we aim to identify molecules able to modulate the activation/activity of aGPCR. For both, phenotyping and structure-function analyses as well as molecule screenings, multiple assays combined with large set ups and numerous samples are required. Thus, an imaging/high content screening system is indispensable. The requested system is flexible and versatile enough to perform all the very different applications in cells as well as in the nematode. It can measure different cellular parameters such as cell size, growth, viability, proliferation, apoptosis, and migration. At the same time, it is able to image and quantify C. elegans individuals in whole-wells and extremely fast. In addition to these relatively low-resolution applications, the system offers the possibility to zoom in on single cells to visualize fluorescence and thus, receptor expression. The system will greatly advance our experimental portfolio and allow for novel advances such as compound screens and large structure-function analyses to gain insights into aGPCR functions.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Bildgebungs-/"High content screening" System
• Instrumentation Group: 5042 Mikroskope für Hochdurchsatz und Screening
• Applicant Institution: Heinrich-Heine-Universität Düsseldorf
• Leader: Professorin Dr. Simone Prömel