Analysis of Cerebrovascular Architecture in Experimental Models of Hypertension and Chronic Neuroendocrine Stress
Authors: Cataldo Noelle , Uhlig Friederike , Johnson Abbie C. , Erdos Benedek 
Published: May 2022

Cerebral small vessel disease and Alzheimer’s disease are leading causes of dementia. Psychological/social stress and associated hypertension are important drivers of brain vascular dysfunction and impaired perfusion. We set out to investigate if chronic neuroendocrine stress and hypertension have adverse effects on cerebrovascular architecture of various brain regions in two rodent models: 1) a rat model where HPA axis activation and hypertension were induced by brain derived neurotrophic factor (BDNF) overexpression in the paraventricular nucleus of the hypothalamus (PVN) for 5 wks, and 2) in a mouse model, where blood pressure and/or corticosterone levels were elevated by subcutaneous infusions of angiotensin II (AngII, 1.5mg/kg/day), adrenocorticotropic hormone (ACTH, 30ug/kg/day) or a combination of AngII and ACTH for 4 weeks. Cerebrovasculature was labeled by transcardiac perfusion of a liposome‑encapsulated fluorescent DiI dye. Brains were fixed in 4% PFA and transparency was increased with the SeeDB method. Vasculature of the hippocampus (HPC), sensory cortex (CTX), and PVN was evaluated using 2‐photon microscopy in 2mm thick coronal brain slices. 3D model of the vasculature was created by Imaris software (Oxford Instruments) yielding parameters such as vascular segment numbers, lengths, diameters, and number of branch points. 3D Euclidean distance mapping (EDM) was used to determine distribution of distances to nearest vessels (DNVs) within the extravascular space as a measure of accessibility to blood supply. We found that vascular densities were markedly elevated in the PVN relative to other brain regions in both rats and mice. In mice, HPC vascular density was significantly lower relative to CTX, whereas in rats, CTX and HPC densities were similar [vascular densities % (rats, mice): PVN:14.6±1.4, 17.0±0.6; CTX: 9.6±0.5, 8.6±0.4; HPC CA: 8.8±0.7, 6.2±0.3; HPC DG: 9.4±0.6, 5.4±0.3]. EDM analysis revealed that the proportion of short (<15μm) vs long DNVs was the highest in the PVN in both rats and mice. Distribution of DNVs shifted to longer distances in the CTX, and the proportion of short vs long DNVs was the lowest in HPC regions indicating an increased vulnerability to hypoperfusion. Interestingly, neuroendocrine stress and hypertension failed to significantly affect vascular densities and DNV statistics in either rats or mice in the analyzed brain regions. However, there was a notable sex difference in distribution of DNVs in HPC CA regions with male rats having a higher proportion of longer distances than females. In addition, analysis of vascular diameter distribution revealed that BDNF treatment decreased vessel diameters in the PVN of both male and female rats, and in the CTX of male, but not female rats. In summary, we found significant region‐specific variability in vascular structure, with the PVN demonstrating a uniquely dense vascular network. Cerebrovascular architecture was fairly resilient to treatments resembling chronic stress‐induced neuroendocrine and cardiovascular responses, but sex differences in HPC vasculature may promote a higher sensitivity to hypoperfusion in male vs female rats.