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

Cited by 34 publications

(32 citation statements)

References 25 publications

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“…-Must be used at low concentrations to avoid in vivo toxicity. (Willis et al, 2010;Saunders et al, 2015;Natarajan et al, 2017;Devraj et al, 2018) Horseradish peroxidase (HRP)…”

Section: Molecular Composition and Regulation Of Tjs At The Bbbmentioning

confidence: 99%

Structure, Function, and Regulation of the Blood-Brain Barrier Tight Junction in Central Nervous System Disorders

et al. 2020

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The blood-brain barrier (BBB) allows the brain to selectively import nutrients and energy critical to neuronal function while simultaneously excluding neurotoxic substances from the peripheral circulation. In contrast to the highly permeable vasculature present in most organs that reside outside of the central nervous system (CNS), the BBB exhibits a high transendothelial electrical resistance (TEER) along with a low rate of transcytosis and greatly restricted paracellular permeability. The property of low paracellular permeability is controlled by tight junction (TJ) protein complexes that seal the paracellular route between apposing brain microvascular endothelial cells. Although tight junction protein complexes are principal contributors to physical barrier properties, they are not static in nature. Rather, tight junction protein complexes are highly dynamic structures, where expression and/or localization of individual constituent proteins can be modified in response to pathophysiological stressors. These stressors induce modifications to tight junction protein complexes that involve de novo synthesis of new protein or discrete trafficking mechanisms. Such responsiveness of BBB tight junctions to diseases indicates that these protein complexes are critical for maintenance of CNS homeostasis. In fulfillment of this vital role, BBB tight junctions are also a major obstacle to therapeutic drug delivery to the brain. There is an opportunity to overcome this substantial obstacle and optimize neuropharmacology via acquisition of a detailed understanding of BBB tight junction structure, function, and regulation. In this review, we discuss physiological characteristics of tight junction protein complexes and how these properties regulate delivery of therapeutics to the CNS for treatment of neurological diseases. Specifically, we will discuss modulation of tight junction structure, function, and regulation both in the context of disease states and in the setting of pharmacotherapy. In particular, we will highlight how these properties can be potentially manipulated at the molecular level to increase CNS drug levels via paracellular transport to the brain.

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“…-Must be used at low concentrations to avoid in vivo toxicity. (Willis et al, 2010;Saunders et al, 2015;Natarajan et al, 2017;Devraj et al, 2018) Horseradish peroxidase (HRP)…”

Section: Molecular Composition and Regulation Of Tjs At The Bbbmentioning

confidence: 99%

Structure, Function, and Regulation of the Blood-Brain Barrier Tight Junction in Central Nervous System Disorders

et al. 2020

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“…was injected 20 mins prior to blood collection and transcardiac perfusion with saline. Brain permeability index (BPI) was calculated comparing fluorescence (relative fluorescence units, RFU) in brain to fluorescence in serum [BPI = (RFU brain/brain weight)/(RFU Serum/serum volume)], and normalized to brain permeability index in control animals (Devraj, Guérit, Macas, & Reiss, 2018). We also used fluorescent microscopy to visualize qualitative differences in sodium fluorescein penetration in brain sections across multiple brain regions (20) (Nikon A1 laser scanning microscope (Devraj et al, 2018;Nikolian et al, 2018).…”

Section: Blood-brain Barrier Permeabilitymentioning

confidence: 99%

Memory deficits in males and females long after subchronic immune challenge

Tchessalova

Tronson

2018

Preprint

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Memory impairments and cognitive decline persist long after recovery from major illness or injury, and correlate with increased risk of later dementia. Here we developed a subchronic peripheral immune challenge model to examine delayed and persistent memory impairments in females and in males. We show that intermittent injections of either lipopolysaccharide or Poly I:C cause memory decline in both sexes that are evident eight weeks after the immune challenge. Importantly, we observed sex-specific patterns of deficits.Females showed impairments in object recognition one week after challenge that persisted for at least eight weeks. In contrast, males had intact memory one week after the immune challenge but exhibited broad impairments in memory tasks including object recognition, and both context and tone fear conditioning several months later. The differential patterns of memory deficits in males and in females were observed without sustained microglial activation or changes in blood-brain barrier permeability. Together, these data suggest that transient neuroimmune activity results in differential vulnerabilities of females and males to memory decline after immune challenge. This model will be an important tool for determining the mechanisms in both sexes that contribute to memory impairments that develop over the weeks and months after recovery from illness. Future studies using this model will provide new insights into the role of chronic inflammation in the pathogenesis of long-lasting memory decline and dementias.

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“…Blood-brain barrier permeability assay. The protocol used to assess blood-brain barrier permeability was adapted from the protocol previously described by Devraj et al (81). Briefly, 100 l 2 mM sodium fluorescein (F6377-100G; Sigma) was administered intraperitoneally to newborn mice at the indicated time points using a 1-ml U-100 syringe (28 gauge by 1/2 inch).…”

Section: Methodsmentioning

confidence: 99%

The Innate Immune Response to Herpes Simplex Virus 1 Infection Is Dampened in the Newborn Brain and Can Be Modulated by Exogenous Interferon Beta To Improve Survival

Giraldo

Wilcox

Longnecker

2020

mBio

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Newborns are particularly susceptible to severe forms of herpes simplex virus 1 (HSV-1) infection, including encephalitis and multisystemic disseminated disease. The underlying age-dependent differences in the immune response that explain this increased susceptibility relative to the adult population remain largely understudied. Using a murine model of HSV-1 infection, we found that newborn mice are largely susceptible to intracranial and intraperitoneal challenge while adult mice are highly resistant. This age-dependent difference correlated with differential basal-level expression of components of innate immune signaling pathways, which resulted in dampened interferon (IFN) signaling in the newborn brain. To explore the possibility of modulating the IFN response in the newborn brain to recapitulate the adult phenotype, we administered exogenous IFN-β in the context of disseminated HSV-1 infection. IFN-β treatment resulted in significantly increased survival and delayed viral neuroinvasion in the newborn. These effects were associated with changes in the type I IFN response in the brain, reduced viral replication in the periphery, and the stabilization of the blood-brain barrier (BBB). Our study reveals important age-dependent differences in the innate immune response to HSV-1 infection and suggests a contribution of the BBB and the brain parenchyma in mediating the increased susceptibility to HSV-1 infection observed in the newborn. These results could provide the basis for potential new therapeutic strategies for life-threatening HSV-1 infection in newborns. IMPORTANCE Herpes simplex virus (HSV) is a ubiquitous human pathogen affecting 50 to 80% of the population in North America and Europe. HSV infection is commonly asymptomatic in the adult population but can result in fatal encephalitis in the newborn. Current treatment with acyclovir has improved mortality in the newborn; however, severe neurologic sequelae are still a major concern following HSV encephalitis. For this reason, there is a critical need to better understand the underlying differences in the immune response between the two age groups that could be used to develop more effective treatments. In this study, we investigated differences in the innate immune response to viral infection in the brains of newborn and adult mice. We found that, similar to humans, newborn mice are more susceptible to HSV infection than the adult. Increased susceptibility was associated with dampened innate immune responses in the newborn brain that could be rescued by administering interferon beta.

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

Cited by 34 publications

References 25 publications

Structure, Function, and Regulation of the Blood-Brain Barrier Tight Junction in Central Nervous System Disorders

Structure, Function, and Regulation of the Blood-Brain Barrier Tight Junction in Central Nervous System Disorders

Memory deficits in males and females long after subchronic immune challenge

The Innate Immune Response to Herpes Simplex Virus 1 Infection Is Dampened in the Newborn Brain and Can Be Modulated by Exogenous Interferon Beta To Improve Survival

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