silicone rubber compound injection and tissue harvest to assess vessel dilatation in neurodegenerative diseases

Dolichoektasie: Vaskuläre Demenz und Alzheimer

Microvascular endothelial cells that line the cerebral capillaries are the main components for forming the blood-brain barrier (BBB)1 which plays a crucial role in regulating what enters or leaves the brain circulation with the periphery. Essential nutrients needed for nervous tissue are permitted to enter the BBB, while some essential amino acids like glutamate are expelled from the brain, as high concentrations can cause permanent neuroexcitatory damage to brain tissue2. Under normal physiological conditions, the BBB limits the amount of plasma proteins like albumin3,4 and prothrombin from entering the brain since those can have detrimental effects5,6,7. Finally, the BBB protects the brain from neurotoxins that are circulating in the periphery, such as xenobiotics from food or the environment1. Overall, damage to brain tissues is irreversible, and aging that correlates with low levels of neurogenesis8 highlights the importance of the BBB in protecting and preventing any factors from accelerating the neurodegenerative process.
In dolichoectasia (or large blood vessel dilatation), a decrease in vessel elasticity is observed, which results in vessels undergoing morphological changes, thus rendering them dysfunctional9 and leading to reduced blood flow in the brain. This reduction in blood flow subsequently diminishes oxygen and glucose supply, ultimately triggering damage to the BBB through the activation of reactive astrocytes10. When the internal elastin lamina of vessels is damaged from dolichoectasia11, repeated stimulation of the vascular endothelial growth factor (VEGF) is necessary for angiogenesis. This can lead to the formation of leaky vessels and ultimately result in pathological angiogenesis, characterized by the development of defective vessels12. During pathological angiogenesis, when blood vessels become defective, a compensatory mechanism appears to restore vessel integrity by upregulating tight junction proteins. However, this process can inadvertently disrupt the BBB when the structural integrity of a blood vessel is lost13. This may occur through further disrupting the BBB and promoting the production of amyloid plaque14. Additionally, leakage from the periphery can cause neuroinflammation15, which results in neuronal degeneration and subsequent memory loss.
Structurally the protection that the BBB provides is through the tight junctions that prevent xenobiotic agents from the blood entering the brain. When permitting certain substances to enter the brain, the BBB mainly does it through two major processes, passive diffusion, or specific channels (like ion channels and transporters)1. In AD, research has demonstrated that a dysfunctional vascular system plays a significant role in the progression of the condition12,13. The formation of amyloid-beta (A&#946;) plaques and neurodegeneration can result from the breakdown of the BBB12,13 and disturbances in cerebral blood flow16. A reduction in cerebral blood flow can be seen in elderly individuals diagnosed with vascular dementia and AD17,18. Damage to the blood-brain barrier (BBB) along with a dysfunctional cerebral blood flow (CBF) can contribute to the increased production of A&#946; concentration in the brain, accompanied by the infiltration of foreign materials from the peripheral circulation19.
To investigate the pathogenesis of neurological diseases like AD and vascular dementia (VaD), models are developed to replicate the disease. In vitro models are extensively used but lack the biological environment for extensive disease modeling like mixed cell population, thus necessitating the importance of in vivo models. Mice are commonly used due to their ease of genetic manipulation in generating human-like properties (e.g., pathology) in disease. With the progression that has been made so far, there is still a continued need for improved models to emulate disease phenotypes like large vessel dilatation and their role in AD. To this end, we saw an opportunity and modified a technique that involved the injection of elastase into the Cisterna magna of mice20,21. Elastase is an enzyme that has been shown to break down elastin in connective tissue22 and in surrounding tight junctions23. The Cisterna magna was chosen as the point of injection due to it being located directly above the circle of Willis, the largest blood vessel in the brain. By injecting elastase into the Cisterna magna, we can compromise the BBB and blood vessels by breaking down the tight junctions and inducing dilatation of blood vessels (circle of Willis)24,25. Combining this technique with the use of an AD mouse model of pathology, for improved understanding of the pathogenesis for the vascular component of AD, can provide valuable insights into the complex interactions and influences between these two distinct pathologies.
Previous studies have demonstrated instances where patients display both the pathological features of AD and VaD, a condition typically referred to as mixed dementia26,27. Thus, understanding the interconnected mechanisms between both conditions can offer a more comprehensive perspective on the progression and manifestation of these neurodegenerative disorders, enhances our comprehension of the underlying mechanisms and potential therapeutic strategies. To this end, we demonstrate the application of elastase in an AD pathology mouse model (AppNL-F) to identify vascular changes.

Autor im Rampenlicht: Modellierung vaskulärer Beiträge zur Alzheimer-Krankheit bei transgenen Mäusen

Ein visueller Ansatz zur Induktion von Dolichoektasie bei Mäusen zur Modellierung der durch große Gefäße vermittelten zerebrovaskulären Dysfunktion

The scope of our research is the model dolichoectasia in a transgenic mouse model to see the maximum impact that vascular changes have on Alzheimer's disease pathology. We are able to show that a single dose of elastase injection into the Cisterna magna of mice is able to cause dilatation up to the three months. Not much is known about individuals who suffer from both vascular dementia and Alzheimer's disease.We hope to bridge this gap by causing vascular changes in these transgenic mouse model to try and understand the pathogenesis of this form of mixed dementia. Previous studies were able to show that dilatation lasts up to two weeks, in our study, we showed that dilatation lasts up to three months with a low mortality during our surgical procedure. Successful dilatation of large blood vessel in a transgenic mouse model provides us with the opportunity to test drugs to prevent or improve vascular changes in Alzheimer's disease.

Injektion von Silikonkautschukmischungen und Gewebeentnahme zur Beurteilung der Gefäßerweiterung bei neurodegenerativen Erkrankungen

silicone-rubber-compound-injection-tissue-harvest-to-assess-vessel

Research

JoVE Journal

Neuroscience

Silicone Rubber Compound Injection and Tissue Harvest to Assess Vessel Dilatation in Neurodegenerative Diseases

47 Views. To begin, place the anesthetized mouse on an absorbent pad or diaper. Pinch the mouse toe to confirm the ceased reflexes. Using surgical tape, secure all limbs with the chest cavity facing upward.Employing tweezers and scissors, carefully open the chest cavity to expose the heart. Insert a 23 gauge blunt needle into the left ventricle of the heart for perfusion. Then mix five milliliters of silicone rubber compound solution, with five milliliters of the diluent in a 50 milliliter tube....