Re-evaluation of Evans Blue dye as a marker of albumin clearance in murine models of acute lung injury

Moitra, Jaideep; Sammani, Saad; Garcia, Joe G.N.

doi:10.1016/j.trsl.2007.03.013

Cited by 106 publications

(103 citation statements)

References 24 publications

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“…for 24 h in anesthetized mice via a 20-gauge catheter, with or without JI-34 (100 μg/kg) or RC-3940-II (100 μg/kg). An Evans Blue dye/Albumin mixture (30 mg/ kg in saline) was injected into the tail vein 2 h before mice were killed, in order to assess vascular leak (39). Lungs free of blood were weighed and snap-frozen in liquid nitrogen.…”

Section: Methodsmentioning

confidence: 99%

Agonist of growth hormone-releasing hormone reduces pneumolysin-induced pulmonary permeability edema

Lucas

Sridhar

Rick

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Aggressive treatment with antibiotics in patients infected with Streptococcus pneumoniae induces release of the bacterial virulence factor pneumolysin (PLY). Days after lungs are sterile, this pore-forming toxin can still induce pulmonary permeability edema in patients, characterized by alveolar/capillary barrier dysfunction and impaired alveolar liquid clearance (ALC). ALC is mainly regulated through Na + transport by the apically expressed epithelial sodium channel (ENaC) and the basolaterally expressed Na + /K + -ATPase in type II alveolar epithelial cells. Because no standard treatment is currently available to treat permeability edema, the search for novel therapeutic candidates is of high priority. We detected mRNA expression for the active receptor splice variant SV1 of the hypothalamic polypeptide growth hormone-releasing hormone (GHRH), as well as for GHRH itself, in human lung microvascular endothelial cells (HL-MVEC). Therefore, we have evaluated the effect of the GHRH agonist JI-34 on PLY-induced barrier and ALC dysfunction. JI-34 blunts PLY-mediated endothelial hyperpermeability in monolayers of HL-MVEC, in a cAMP-dependent manner, by means of reducing the phosphorylation of myosin light chain and vascular endothelial (VE)-cadherin. In human airway epithelial H441 cells, PLY significantly impairs Na + uptake, but JI-34 restores it to basal levels by means of increasing cAMP levels. Intratracheal instillation of PLY into C57BL6 mice causes pulmonary alveolar epithelial and endothelial hyperpermeability as well as edema formation, all of which are blunted by JI-34. These findings point toward a protective role of the GHRH signaling pathway in PLY-induced permeability edema.pneumonia | cyclic AMP

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Section: Methodsmentioning

confidence: 99%

Agonist of growth hormone-releasing hormone reduces pneumolysin-induced pulmonary permeability edema

Lucas

Sridhar

Rick

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Data were stored and analyzed with an SPSS package (SPSS Inc., Chicago, III) as described previously. 41 To detect a phenotype, type I sums of squares (for fixed effects or model I analysis of variance) were computed for a 4-way factorial design in which genotype, treatment, age, and sex were considered as independent contributors (factors) to observed variances of means. 49 However, as SPSS reports output of such multiway analysis of variance (ANOVA) in the form of multiple contrasts alone, the fixed factors (age, sex, genotype, and treatment) were collapsed into 24 permutations (wild-type untreated young female, wild-type LPS-treated young female, transgenic ventilated old male, etc) and analyzed in 1-way (single factor classification analysis of variance) ANOVA terms to facilitate visualization of significant differences between groups.…”

Section: Discussionmentioning

confidence: 99%

“…We have assumed that an increase in blood proteins per unit mass of lung parenchyma is a more useful indicator of the existence of a paracellular gap, an idea similar to the popular Evans Blue dye leakage assay. 41 An advantage of this approach is the evaluation of endothelial but not epithelial gap formation-the size of the gap is approximated by the size of the trapped protein regardless of the presence/absence of these proteins in the BAL fluid.…”

Section: Discussionmentioning

confidence: 99%

“…Despite the high success rate of embryo survival in hybrid strains, 39 we chose the B6 strain rather than a hybrid strain primarily because inbred strains eliminate ambiguity caused by different genetic backgrounds and segregating markers in the progeny, and secondarily because we are familiar with this strain in terms of their ALI/VALI responses. [ 40 ], [ 41 ] and [ 42 ] The transgene specific primers were J9 (forward; 5′-GTGGCTCCCCTCTCCCCTCCTG-3′) and J30 (reverse; 5′-CTTGGCGGTGGCTCCTTCTTTGAT-3′), which gave a 319-bp product. Mouse β-globin gene-specific primers J18 (forward; 5′-GGCAGCTCACAAGAAGAAGTTGGG-3′) and J19 (reverse; 5′-ATCAAAGTACCGCTGGGTCCAAGG-3′) that amplify a 138-bp band were included in all reactions as positive control.…”

Section: Generation Of Transgenic Micementioning

confidence: 99%

“…The LPS-induced ALI model has been described previously. [ 40 ] and [ 41 ] Briefly, mice were anesthetized with intraperitoneal ketamine (150 mg/kg) and acetylpromazine (15 mg/kg) before the exposure of the trachea and the right internal jugular vein via neck incision. Sterile water (pH 7.0) or LPS (2.5 mg/kg in 50 μL of the same vehicle) were instilled intratracheally via a 20-gauge catheter.…”

Section: Murine Model Of Lps-induced Injurymentioning

confidence: 99%

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A transgenic mouse with vascular endothelial over-expression of the non-muscle myosin light chain kinase-2 isoform is susceptible to inflammatory lung injury: role of sexual dimorphism and age

Moitra

Evenoski

Sammani

et al. 2008

Translational Research

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We have generated genetically engineered mice that are uniquely susceptible to lipopolysaccharide (LPS)-induced and mechanical ventilation-induced lung injury in a sex-specific and age-specific manner. These mice express a nonmuscle isoform of the myosin light chain kinase gene (nmMLCK2) targeted to the endothelium. Homozygous mice have significantly reduced fecundity and litter survival until weaning, and they are initially growth delayed but eventually exceed the size of wildtype littermates. Mice at all ages show increased protein transport across the lung barrier; however, the phenotype is most discernible in 8-12-week-old male mice. When subjected to a clinically relevant LPS-induced lung injury model, 8-12-week-old young females and 30-36-week-old males seem to be the most significantly injured group. In contrast, 30-36-week-old males remain the most significantly injured group when mechanically ventilated at high tidal volumes, which is a clinically relevant model of mechanical stress lung injury. These data reveal that nmMLCK2 overexpression in the endothelium exacerbates lung injury in vivo in a sexually dimorphic and age-dependent manner.Actin microfilaments generate force by virtue of their ability to contract away from their site of attachment to the plasma membrane, and they do so via traction of the myosin light chain head groups attached to actin filaments by cross-bridges. The cross-bridge pulling action of myosin involves a conformational change in the molecule initiated by energetic phosphorylation of Ser19 on the myosin light chains. 1 The key enzyme involved in this phosphorylation event is myosin light chain kinase (MLCK), which exists in different tissues, and it may be characterized broadly as skeletal/cardiac, smooth muscle, and nonmuscle myosin light chain kinase (nmMLCK) isoforms. Although biochemical evidence for the existence of a Ca 2+ /calmodulin-dependent nonmuscle isoform in cultured endothelial cell (EC) was suggested earlier, [ 2 ] and [ 3 ] the controversy as to whether this enzymatic activity is identical to the 110-kDa protein found abundantly in smooth muscle preparations was resolved when we provided conclusive proof of a larger MLCK protein isoform (210 kDa) by cloning the gene (MYLK) from a human EC-derived cDNA library. 4 The gene was subsequently mapped to chromosome 3q21. 5

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Cerebral Microvascular Injury Induced by Lag3‐Dependent α‐Synuclein Fibril Endocytosis Exacerbates Cognitive Impairment in a Mouse Model of α‐Synucleinopathies

et al. 2023

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The pathological accumulation of α‐synuclein (α‐Syn) and the transmission of misfolded α‐Syn underlie α‐synucleinopathies. Increased plasma α‐Syn levels are associated with cognitive impairment in Parkinson's disease, multiple system atrophy, and dementia with Lewy bodies, but it is still unknown whether the cognitive deficits in α‐synucleinopathies have a common vascular pathological origin. Here, it is reported that combined injection of α‐Syn preformed fibrils (PFFs) in the unilateral substantia nigra pars compacta, hippocampus, and cerebral cortex results in impaired spatial learning and memory abilities at 6 months post‐injection and that this cognitive decline is related to cerebral microvascular injury. Moreover, insoluble α‐Syn inclusions are found to form in primary mouse brain microvascular endothelial cells (BMVECs) through lymphocyte‐activation gene 3 (Lag3)‐dependent α‐Syn PFFs endocytosis, causing poly(ADP‐ribose)‐driven cell death and reducing the expression of tight junction proteins in BMVECs. Knockout of Lag3 in vitro prevents α‐Syn PFFs from entering BMVECs, thereby reducing the abovementioned response induced by α‐Syn PFFs. Deletion of endothelial cell‐specific Lag3 in vivo reverses the negative effects of α‐Syn PFFs on cerebral microvessels and cognitive function. In short, this study reveals the effectiveness of targeting Lag3 to block the spread of α‐Syn fibrils to endothelial cells in order to improve cognition.

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Re-evaluation of Evans Blue dye as a marker of albumin clearance in murine models of acute lung injury

Cited by 106 publications

References 24 publications

Agonist of growth hormone-releasing hormone reduces pneumolysin-induced pulmonary permeability edema

Agonist of growth hormone-releasing hormone reduces pneumolysin-induced pulmonary permeability edema

A transgenic mouse with vascular endothelial over-expression of the non-muscle myosin light chain kinase-2 isoform is susceptible to inflammatory lung injury: role of sexual dimorphism and age

Cerebral Microvascular Injury Induced by Lag3‐Dependent α‐Synuclein Fibril Endocytosis Exacerbates Cognitive Impairment in a Mouse Model of α‐Synucleinopathies

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