Influence of Nanoparticle Exposure on Nervous System Development in Zebrafish Studied by Means of Light Sheet Fluorescence Microscopy

Abstract

Zebrafish has a remarkable similarity in the molecular signaling processes, cellular structure, anatomy and physiology to other higher order vertebrates, making it an excellent vertebrate model organism (1). Recently, zebrafish has been used for neurotoxicity screening of numerous nanomaterials with a focus on the developmental effects due to the possibility of in vivo visualization of specific neurons and axon tracts by injecting dyes in live animals as well in fixed ones (2). Here we propose Light Sheet Fluorescence Microscopy (LSFM) (3) (4) to perform neurotoxicity studies, in order to study the nervous system architecture and to image 3D structures in the brain of live larvae during the development. It is worth noting that the 3D imaging capabilities and the fast imaging speed provided by LSFM, combined with the low photo damage levels indicate this imaging technique as an ‘election’ method for imaging neurons distribution during embryo development. Specifically, to evaluate the neurotoxicity of nanoparticles on zebrafish, we characterized the apoptotic processes and axon tract disruption in the brain during development, that correlated cerebral malformations and anomalies in the behavior. Here, we report about the correlation of nanoparticle exposure with the neuronal number and related brightness using light sheet fluorescence microscopy. LSFM is also suitable for 3D super resolved investigations (5).(1)Truong L. et al., Methods Mol Biol; 691: 271-279 (2011)(2)Parn C. et al., J Pharmacol Toxicol Methods; 55(1):103-112 (2007)(3)Huisken J. et al., Science; 305: 1007-1009 (2004)(4)Stelzer E. H. K.et al., Nature Methods; 12: 23-26 (2015)(5)Cella Zanacchi et al., Nature Methods; 8:1047-1049 (2011)