An &lt;em&gt;In Vivo&lt;/em&gt; Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Devraj, Kavi; Guérit, Sylvaine; Macas, Jakranka; Reiss, Yvonne

doi:10.3791/57038-v

Cited by 4 publications

(3 citation statements)

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“…To determine small molecule leak at the pre-plaque stages, we used methods that were adapted from [ 55 ]. Briefly, 2- to 3-month-old wildtype and APP/PS1 mice were each injected with 100 mg/kg body weight of unconjugated biotin.…”

Section: Methodsmentioning

confidence: 99%

Vascular senescence and leak are features of the early breakdown of the blood–brain barrier in Alzheimer’s disease models

Ting,

Coleman,

Kim

et al. 2023

GeroScience

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Alzheimer’s disease (AD) is an age-related disease, with loss of integrity of the blood–brain barrier (BBB) being an early feature. Cellular senescence is one of the reported nine hallmarks of aging. Here, we show for the first time the presence of senescent cells in the vasculature in AD patients and mouse models of AD. Senescent endothelial cells and pericytes are present in APP/PS1 transgenic mice but not in wild-type littermates at the time of amyloid deposition. In vitro, senescent endothelial cells display altered VE-cadherin expression and loss of cell junction formation and increased permeability. Consistent with this, senescent endothelial cells in APP/PS1 mice are present at areas of vascular leak that have decreased claudin-5 and VE-cadherin expression confirming BBB breakdown. Furthermore, single cell sequencing of endothelial cells from APP/PS1 transgenic mice confirms that adhesion molecule pathways are among the most highly altered pathways in these cells. At the pre-plaque stage, the vasculature shows significant signs of breakdown, with a general loss of VE-cadherin, leakage within the microcirculation, and obvious pericyte perturbation. Although senescent vascular cells were not directly observed at sites of vascular leak, senescent cells were close to the leak area. Thus, we would suggest in AD that there is a progressive induction of senescence in constituents of the neurovascular unit contributing to an increasing loss of vascular integrity. Targeting the vasculature early in AD, either with senolytics or with drugs that improve the integrity of the BBB may be valid therapeutic strategies.

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Section: Methodsmentioning

confidence: 99%

Vascular senescence and leak are features of the early breakdown of the blood–brain barrier in Alzheimer’s disease models

Ting,

Coleman,

Kim

et al. 2023

GeroScience

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“…To further study the BBB penetration activity of βγ-CAT, the in vivo assay was performed according to previously described methods [ 62 ]. Briefly, C57 mice were divided into 2 groups.…”

Section: Methodsmentioning

confidence: 99%

A Pore Forming Toxin-like Protein Derived from Chinese Red Belly Toad Bombina maxima Triggers the Pyroptosis of Hippomal Neural Cells and Impairs the Cognitive Ability of Mice

Wang

Lee

et al. 2023

Toxins

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Toxin-like proteins and peptides of skin secretions from amphibians play important physiological and pathological roles in amphibians. βγ-CAT is a Chinese red-belly toad-derived pore-forming toxin-like protein complex that consists of aerolysin domain, crystalline domain, and trefoil factor domain and induces various toxic effects via its membrane perforation process, including membrane binding, oligomerization, and endocytosis. Here, we observed the death of mouse hippocampal neuronal cells induced by βγ-CAT at a concentration of 5 nM. Subsequent studies showed that the death of hippocampal neuronal cells was accompanied by the activation of Gasdermin E and caspase-1, suggesting that βγ-CAT induces the pyroptosis of hippocampal neuronal cells. Further molecular mechanism studies revealed that the pyroptosis induced by βγ-CAT is dependent on the oligomerization and endocytosis of βγ-CAT. It is well known that the damage of hippocampal neuronal cells leads to the cognitive attenuation of animals. The impaired cognitive ability of mice was observed after intraperitoneal injection with 10 μg/kg βγ-CAT in a water maze assay. Taken together, these findings reveal a previously unknown toxicological function of a vertebrate-derived pore-forming toxin-like protein in the nerve system, which triggers the pyroptosis of hippocampal neuronal cells, ultimately leading to hippocampal cognitive attenuation.

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“…Frozen brains were sectioned at 12 µm-thick sections -1.5 to -2.1 mm from bregma and fixed with 1% PFA. Fixed brains were stained with rabbit anti-CD31 (BD bioscience, Catalog#: 553370, Isotype: Rat IgG2a, Lot#: B313252, 0.5mg/ml, clone MEC 13.3, 1:100) for 1.5 hours at room temperature [ 45 ]. Brains were stained with secondary anti-rabbit cy5 (Biotium, Catalog#:89138-520, Lot#18C1120, 1:500) labeled for 1 hour at room temperature.…”

Section: Methodsmentioning

confidence: 99%

PNA5, A Novel Mas Receptor Agonist, Improves Neurovascular and Blood-Brain-Barrier Function in a Mouse Model of Vascular Cognitive Impairment and Dementia

2023

Aging dis

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It is well established that decreased brain blood flow, increased reactive oxygen species production (ROS), and pro-inflammatory mechanisms accelerate neurodegenerative disease progressions, including vascular cognitive impairment and dementia (VCID). Previous studies in our laboratory have shown that our novel glycosylated Angiotensin-(1-7) Mas receptor agonist PNA5 reverses cognitive deficits, decreases ROS production, and inhibits inflammatory cytokine production in our preclinical mouse model of VCID that is induced by chronic heart failure (VCID-HF). In the present study, the effects of VCID-HF and treatment with PNA5 on microglia activation, blood-brain-barrier (BBB) integrity, and neurovascular coupling were assessed in our mouse model of VCID-HF. Three-month-old male C57BL/6J mice were subjected to myocardial infarction (MI) to induce heart failure for four weeks and then treated with subcutaneous injections of extended-release PNA5. Microglia activation, BBB permeability, cerebral perfusion, and neurovascular coupling were assessed. Results show that in our VCID-HF model, there was an increase in microglial activation and recruitment within the CA1 and CA3 regions of the hippocampus, a disruption in BBB integrity, and a decrease in neurovascular coupling. Treatment with PNA5 reversed these neuropathological effects of VCID-HF, suggesting that PNA5 may be an effective disease-modifying therapy to treat and prevent VCID. This study identifies potential mechanisms by which heart failure may induce VCID and highlights the possible mechanisms by which treatment with our novel glycosylated Angiotensin-(1-7) Mas receptor agonist, PNA5, may protect cognitive function in our model of VCID.

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An <em>In Vivo</em> Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Cited by 4 publications

References 0 publications

Vascular senescence and leak are features of the early breakdown of the blood–brain barrier in Alzheimer’s disease models

Vascular senescence and leak are features of the early breakdown of the blood–brain barrier in Alzheimer’s disease models

A Pore Forming Toxin-like Protein Derived from Chinese Red Belly Toad Bombina maxima Triggers the Pyroptosis of Hippomal Neural Cells and Impairs the Cognitive Ability of Mice

PNA5, A Novel Mas Receptor Agonist, Improves Neurovascular and Blood-Brain-Barrier Function in a Mouse Model of Vascular Cognitive Impairment and Dementia

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