Simultaneous, live imaging of cortical spreading depression and associated cerebral blood flow changes, by combining voltage-sensitive dye and laser speckle contrast methods.
Cortical spreading depression (i.e. waves of cellular depolarization, CSD) causes the aura symptoms in classical migraine, and may contribute to delayed cellular damage after an ischemic or traumatic insult to the brain. In the latter cases, secondary neuronal injury may be worsened by some of the cerebral blood flow (CBF) changes that are associated with CSD. Here, we describe a new method for the simultaneous, live imaging of local cellular depolarization and CBF changes (i.e. two variables with well-defined and important biological significance), through a closed cranial window prepared in anesthetized rats. This novel experimental strategy was validated by imaging the changes associated with CSD elicited by application of high K(+) medium on the cortical surface. CSD was visualized directly by using a fluorescent voltage-sensitive (VS) dye, whereas laser speckle contrast (LSC) imaging allowed the visualization of the corresponding CBF changes. In addition to the high temporal and spatial resolution of VS dye and LSC imaging, their novel combination allows to determine how CBF changes relate to a heterogeneous and evolving pattern of cellular depolarization, in any area of interest of the cortical region under study. This methodological development is especially pertinent and timely for investigations into the peri-lesion depolarizations that occur in models of focal brain injury, situations where their site of spontaneous elicitation and propagation pattern cannot be predicted. It should also help advance our knowledge in epilepsy, CBF pharmacology, and neurovascular coupling under normal and pathophysiological conditions.
Obrenovitch
Tihomir P
TP
University of Bradford, UK. t.obrenovitch@bradford.ac.uk
Chen
Shangbin
S
Farkas
Eszter
E
D000818:Animals; D001783:Blood Flow Velocity / Q000502:physiology*; D001921:Brain / Q000098:blood supply / Q000502:physiology*; D001931:Brain Mapping / Q000379:methods*; D002560:Cerebrovascular Circulation / Q000502:physiology*; D013181:Cortical Spreading Depression; D007090:Image Interpretation, Computer-Assisted / Q000379:methods; D017078:Laser-Doppler Flowmetry / Q000379:methods*; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley