In addition to standard in vitro biochemical, molecular cloning and multiple imaging approaches, the Washbourne Lab uses a combination of three principle model systems: cell cultures, developing and adult zebrafish and transgenic mice. We use the strong points of each model system to further our understanding of the molecular and cellular interactions during synapse formation:
Primary neuronal cultures and cell lines – We take advantage of the unparalleled access of neuronal and non-neuronal cultures for cellular manipulations, electrophysiology and synaptic labeling (Hoy et al. 2009, Barrow et al. 2009).
Developing zebrafish embryos – Zebrafish embryos and larvae are optically clear, lending themselves to time-lapse imaging of subcellular structures in vivo. We have pioneered the use of zebrafish for the study of the development of a central glutamatergic synapses in the developing spinal cord of a living vertebrate (Pietri et al. 2009; Easley-Neal et al. 2013). We have characterized key genes, synaptic markers and basic anatomy to provide a platform with which to now dissect the molecular mechanisms of synapse formation in vivo (Pietri et al. 2008; Davey et al. 2010).
Transgenic mice – The mouse provides a combination of genetic control, together with electrophysiological and behavioral analysis critical for the analysis of the consequences of perturbations of synapses. Furthermore, we are preparing studies to test the possibility of genetic rescue of an autism phenotype in mice, demonstrating a high level of clinical relevance of our mouse behavioral research (Hoy et al. 2013).