An artery-specific fluorescent dye for studying neurovascular coupling

Shen, Zhiming; Lu, Zhongyang; Chhatbar, Pratik Y.; O’Herron, Philip; Kara, Prakash

doi:10.1038/nmeth.1857

Cited by 150 publications

(175 citation statements)

References 22 publications

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“…For example, AECs expressed high levels of DLL4, which can activate the Notch pathway and promote cardiac regeneration (40)(41)(42)(43)(44)(45). Another mechanism may be high NO production as NO plays a pivotal role in ischemic protection (46). Results suggest that these AECs could have therapeutic value by themselves or in more complex tissue constructs.…”

Section: Discussionmentioning

confidence: 99%

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Functional characterization of human pluripotent stem cell-derived arterial endothelial cells

Zhang

Chu

Hou

et al. 2017

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

107

106

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Here, we report the derivation of arterial endothelial cells from human pluripotent stem cells that exhibit arterial-specific functions in vitro and in vivo. We combine single-cell RNA sequencing of embryonic mouse endothelial cells with an EFNB2-tdTomato/ EPHB4-EGFP dual reporter human embryonic stem cell line to identify factors that regulate arterial endothelial cell specification. The resulting xeno-free protocol produces cells with gene expression profiles, oxygen consumption rates, nitric oxide production levels, shear stress responses, and TNFα-induced leukocyte adhesion rates characteristic of arterial endothelial cells. Arterial endothelial cells were robustly generated from multiple human embryonic and induced pluripotent stem cell lines and have potential applications for both disease modeling and regenerative medicine.arterial endothelial cells | arterial-specific functions | myocardial infarction | single-cell RNA-seq | human pluripotent stem cell differentiation

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Section: Discussionmentioning

confidence: 99%

“…AECs and VECs were stained with human-specific CD31 antibody. Alexa Fluor 647 Hydrazide (1 μg/mL) was used to label the elastin produced by arteries (46). hCD31, anti-human CD31 antibody.…”

Section: Methodsmentioning

confidence: 99%

Functional characterization of human pluripotent stem cell-derived arterial endothelial cells

Zhang

Chu

Hou

et al. 2017

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

107

106

View full text Add to dashboard Cite

show abstract

“…To visualize vessels, a 0.1 ml bolus of 5% (w/v) Texas Red dextran (70 kDA; Invitrogen) in 0.9% NaCl was injected into the tail vein. In some experiments, arteriole visualization was conducted through topical application of Alexa 633 as previously reported (Shen et al, 2012). The skin covering the cranium above the somatosensory cortex was removed and a custom-made 1.3-cm-diameter plastic ring was glued with dental acrylic cement (Co-Oral-Ite Dental) to stabilize the head during craniotomy and imaging with a headholder attached to a baseplate.…”

Section: Animalsmentioning

confidence: 99%

Astrocyte Contributions to Flow/Pressure-Evoked Parenchymal Arteriole Vasoconstriction

et al. 2015

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Basal and activity-dependent cerebral blood flow changes are coordinated by the action of critical processes, including cerebral autoregulation, endothelial-mediated signaling, and neurovascular coupling. The goal of our study was to determine whether astrocytes contribute to the regulation of parenchymal arteriole (PA) tone in response to hemodynamic stimuli (pressure/flow). Cortical PA vascular responses and astrocytic Ca 2ϩ dynamics were measured using an in vitro rat/mouse brain slice model of perfused/pressurized PAs; studies were supplemented with in vivo astrocytic Ca 2ϩ imaging. In vitro, astrocytes responded to PA flow/pressure increases with an increase in intracellular Ca 2ϩ . Astrocytic Ca 2ϩ responses were corroborated in vivo, where acute systemic phenylephrine-induced increases in blood pressure evoked a significant increase in astrocytic Ca 2ϩ . In vitro, flow/pressure-evoked vasoconstriction was blunted when the astrocytic syncytium was loaded with BAPTA (chelating intracellular Ca 2ϩ ) and enhanced when high Ca 2ϩ or ATP were introduced to the astrocytic syncytium. Bath application of either the TRPV4 channel blocker HC067047 or purinergic receptor antagonist suramin blunted flow/pressure-evoked vasoconstriction, whereas K ϩ and 20-HETE signaling blockade showed no effect. Importantly, we found TRPV4 channel expression to be restricted to astrocytes and not the endothelium of PA. We present evidence for a novel role of astrocytes in PA flow/pressure-evoked vasoconstriction. Our data suggest that astrocytic TRPV4 channels are key molecular sensors of hemodynamic stimuli and that a purinergic, glial-derived signal contributes to flow/pressure-induced adjustments in PA tone. Together our results support bidirectional signaling within the neurovascular unit and astrocytes as key modulators of PA tone.

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“…The body temperature of the mice was kept constant at 37 C during all procedures. The MFPs [50 mmol/L in 50 mL of saline containing 1% of poly(ethylene glycol)4600(PEG) and 0.1% of Tween 20] were intradermally injected either in the left forepaw, for normal and inflammation model imaging, or in both the forepaws for tumor model imaging.…”

Section: In Vivo Fluorescence Imaging Of Lymph Nodementioning

confidence: 99%

“…Completely defined synthetic vital dyes have considerable potential to rapidly label cells, subcellular compartments, and molecular effectors in living objects due to their diverse biologic properties caused by skeletal and stereochemical complexities (37,38). In this study, we report the development of a macrophage-specific fluorescent probe (MFP) using a diversity-oriented synthesis (39,40) for intraoperative lymph node staging.…”

Section: Introductionmentioning

confidence: 99%

A Macrophage-Specific Fluorescent Probe for Intraoperative Lymph Node Staging

et al. 2014

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Successful identification of nodal metastases in patients with cancer is crucial to prescribe suitable treatment regimens that can improve recurrence-free survival. Although some new imaging technologies for nodal staging have been developed, such as nanoparticle-enhanced MRI and quantum-dot-based fluorescence imaging, sound technologies for intraoperative differentiation of metastatic and inflamed lymph nodes remain lacking. In this study, we illustrate the feasibility of using a macrophage-specific fluorescent probe (MFP) to visualize sentinel lymph nodes during surgery, highlighting abnormalities related to inflammation and tumor infiltration with signal enhancement and reduction methods using this technology. MFP was identified by high-throughput screening of fluorescent small-molecule libraries synthesized with a diversity-oriented approach. It selectively visualized monocyte and macrophage cell populations in vitro, by live-cell imaging and flow cytometry, as well as in vivo, for imaging-guided surgery. Collectively, this study provides preclinical proof of concept for an intraoperative imaging platform to accurately assess lymph node status, eliminating the need for invasive nodal dissections that can contribute to complications of cancer therapy. Cancer Res; 74(1); 44-55. Ó2013 AACR.

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An artery-specific fluorescent dye for studying neurovascular coupling

Cited by 150 publications

References 22 publications

Functional characterization of human pluripotent stem cell-derived arterial endothelial cells

Functional characterization of human pluripotent stem cell-derived arterial endothelial cells

Astrocyte Contributions to Flow/Pressure-Evoked Parenchymal Arteriole Vasoconstriction

A Macrophage-Specific Fluorescent Probe for Intraoperative Lymph Node Staging

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