Protocol for mouse vascular dementia model and evaluation of progressive tissue damage in subcortical white matter and adjacent cortex.

Here, we present a protocol for generating a vascular dementia (VaD) mouse model through stereotaxic microinjection of a vasoconstrictor into the subcortical white matter (WM). We describe steps for preparing the N5-(1-iminoethyl)-L-ornithine (L-NIO) solution, surgical procedures, and immunohistochemistry. We then detail procedures for confocal imaging and image analysis using ImageJ and Imaris software to evaluate key pathological features of human VaD, including vessel dilation, demyelination, glial reactivity, neuronal damage, subcortical WM atrophy, and ventriculomegaly. For complete details on the use and execution of this protocol, please refer to Tian et al.1.