Project 8 (IEM-HAS)
Project 8: Interaction of NP with neural stem- and tissue-type cells
Fellow: Murali Kumarasamy (ESR8), 36 months
Tutor: Dr. Emilia Madarasz (IEM-HAS)
The vast majority of cells in the central nervous tissue are protected from extraneural chemical/physical insults by multiple (blood-brain, blood-liquor, liquor-tissue) barriers. The extracellular environment of the brain tissue is also controlled by potent glial uptake mechanisms. While “natural” load of NP (from dust, water, etc.) has not been shown to endanger neural functions, we should be aware of potential harm caused by artificially increased amounts of NP in the environment or by direct load of NP as diagnostic probe-materials or by the intended use of nanodevices. The Cellular and Developmental Neurobiology Unit of IEM-HAS provides training and research facilities to investigate neural cell- and tissue reactions in response to NP loading. An ESR will be acquainted and trained in basic neural cell biology methods and in technologies needed for congruent research focused on selected aspects of “neural nanosafety”.
Primary cultures of rat neurons and/or glial cells, neural stem cell clones and in vitro cell differentiation model systems will be used to investigate:
- reactions of various neural cell types to NP-loading (viability, proliferation and differentiation assays; electrophysiological responses);
- mechanisms of permeation of NP through the membranes of different neural cell types (sensor-assays on endocytosis; electronmicroscopy);
- changes in intercellular (neuronal-neuronal, neuronal-glial) interactions in the presence of functionalized or plain NP (cadherin distribution, gap junction communication, ion-barrier characteristics);
- changes in cellular and neuronal process motility (time-lapse video microscopy; process-elongation assays);
- secretory responses of neural cells in the presence of different NP (immune-OWLS assays, cell-blotting) in the presence of different NP.
Besides in vitro analyses, the potential effects of NP on the developing and mature central nervous tissue will be investigated by surgical and histological/immunocytochemical methods in early stages of the embryonic tissue genesis in rodents (Wistar rats), when the neural tissue is not yet protected by specialised barriers, and in mature animals, when NP are expected to accumulate at blood-brain, blood-liquor and internal cellular barrier interfaces. The fellow will get an insight in the cell-type and differentiation-stage dependent variations of responses to NP loading. Also, to evaluate crucial thresholds of NP toxicity, dose-response relations will be determined for cell-type specific reactions of different neural tissue cells. The main goal will be to explore the most vulnerable neural cell types and neural reactions in conditions of NP exposure. Results will be compared to those obtained on non-neural cells and barriers in partner Laboratories. Nanomaterial loading without damaging consequences will be considered for cell labelling and transport-study purposes. Secondments to partners 5 (comparison of vertebrate and invertebrate systems), 9 (evolution of NP in complex systems), 10 (novel cell-based assays).
In WP1 (tasks 1.1, 1.2 and 1.3), WP2 (task 2.5) and WP3 (task 3.1).