TY - JOUR T1 - The within-subject application of diffusion tensor MRI and CLARITY reveals brain structural changes in <em>Nrxn2</em> deletion mice JF - bioRxiv DO - 10.1101/300806 SP - 300806 AU - Eleftheria Pervolaraki AU - Adam L. Tyson AU - Francesca Pibiri AU - Steven L. Poulter AU - Amy C. Reichelt AU - R. John Rodgers AU - Steven J. Clapcote AU - Colin Lever AU - Laura C. Andreae AU - James Dachtler Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/01/29/300806.abstract N2 - Background Of the many genetic mutations known to increase the risk of autism spectrum disorder, a large proportion cluster upon synaptic proteins. One such family of presynaptic proteins are the neurexins (NRXN), and recent genetic and mouse evidence has suggested a causative role for NRXN2 in generating altered social behaviours. Autism has been conceptualised as a disorder of atypical connectivity, yet how single-gene mutations affect such connectivity remains under-explored. To attempt to address this, we have developed a quantitative analysis of microstructure and structural connectivity leveraging diffusion tensor MRI (DTI) with high-resolution 3D imaging in optically cleared (CLARITY) brain tissue in the same mouse, applied here to the Nrxn2α knockout (KO) model.Methods Fixed brains of Nrxn2α KO mice underwent DTI using 9.4T MRI, and diffusion properties of socially-relevant brain regions were quantified. The same tissue was then subjected to CLARITY to immunolabel axons and cell bodies, which were also quantified.Results DTI revealed decreases in fractional anisotropy and increases in apparent diffusion coefficient in the amygdala (including the basolateral nuclei), the anterior cingulate cortex, the orbitofrontal cortex and the hippocampus. Radial diffusivity of the anterior cingulate cortex and orbitofrontal cortex was significantly increased in Nrxn2α KO mice, as were tracts between the amygdala and the orbitofrontal cortex. Using CLARITY, we find significantly altered axonal orientation in the amygdala, orbitofrontal cortex and the anterior cingulate cortex, which was unrelated to cell density.Conclusions Our findings demonstrate that deleting a single neurexin gene (Nrxn2α) induces atypical structural connectivity within socially-relevant brain regions. More generally, our combined within-subject DTI and CLARITY approach presents a new, more sensitive method of revealing hitherto undetectable differences in the autistic brain.ACC: anterior cingulate cortexAD: axial diffusivityADC: apparent diffusion coefficientASD: autism spectrum disorderBLA: basolateral amygdalaCLARITY: optically cleared brain tissueDTI: diffusion tensor imagingFA: fractional anisotropyOFC: orbitofrontal cortexNrxn2: neurexin IIRD: radial diffusivityROI: region of interest ER -