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 • Procedures for generating a VaD mouse model via stereotaxic microinjection • Steps for quantifying core pathological features including cell-type-specific changes • Instructions for longitudinally tracking pathological progression of disease stages • Guidance on comprehensive analysis including immunohistochemistry and image processing Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.