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Quantitative measurement of lymphatic function in mice by noninvasive near-infrared imaging of a peripheral vein
Steven T. Proulx, … , Jean-Christophe Leroux, Michael Detmar
Steven T. Proulx, … , Jean-Christophe Leroux, Michael Detmar
Published January 12, 2017
Citation Information: JCI Insight. 2017;2(1):e90861. https://doi.org/10.1172/jci.insight.90861.
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Resource and Technical Advance Vascular biology

Quantitative measurement of lymphatic function in mice by noninvasive near-infrared imaging of a peripheral vein

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Abstract

Optical imaging methods have been developed to measure lymphatic function in skin; however, the lymphatic system of many organs is not accessible to this technology. Since lymphatic transport of macromolecules from any organ proceeds to the blood circulation, we aimed to develop a method that can measure lymphatic function by monitoring the fluorescence in a superficial vein of an interstitially injected tracer. We selected a 40-kDa PEGylated near-infrared dye conjugate, as it showed lymphatic system–specific uptake and extended circulation in blood. Lymphatic transport to blood from subcutaneous tissue required a transit time before signal enhancement was seen in blood followed by a steady rise in signal over time. Increased lymphatic transport was apparent in awake mice compared with those under continuous anesthesia. The methods were validated in K14-VEGFR-3-Fc and K14-VEGF-C transgenic mice with loss and gain of lymphatic function, respectively. Reduced lymphatic transport to blood was also found in aged mice. The technique was also able to measure lymphatic transport from the peritoneal cavity, a location not suitable for optical imaging. The method is a promising, simple approach for assessment of lymphatic function and for monitoring of therapeutic regimens in mouse models of disease and may have potential for clinical translation.

Authors

Steven T. Proulx, Qiaoli Ma, Diana Andina, Jean-Christophe Leroux, Michael Detmar

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Figure 4

Limited lymphatic transport in K14-VEGFR-3-Fc mice compared with WT littermates.

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Limited lymphatic transport in K14-VEGFR-3-Fc mice compared with WT litt...
Twelve-week-old female K14-VEGFR-3-Fc or WT littermates were s.c. injected with 5 μl of 200 μM 40-kDa PEG–IRDye800 conjugate (P40D800) in the dorsal aspect of the right rear paw. (A) Representative saphenous vein signal in a WT mouse 60 minutes after s.c. injection of P40D800 during anesthetized conditions with the paw massaged for 10 seconds every 5 minutes from t = 5 to 50 minutes. (B) Representative saphenous vein signal in a K14-VEGFR-3-Fc mouse under the same conditions. Scale bars: 500 μm. (C) Saphenous vein signal enhancement plots of K14-VEGFR-3-Fc and WT mice (n = 4 each). Dashed lines indicate SD. (D) Quantification of the fluorescent signal enhancement at t = 60 minutes. (E) Linear slope of signal enhancement from t = 30 to 45 minutes. (F) Time of arrival of tracer to the blood circulation (set at a threshold of 100 counts of signal enhancement). Three of 4 mice in the K14-VEGFR-3-Fc group did not reach this threshold. (G) Quantification of fluorescent signal from a 40-kDa PEG–IRDye680 conjugate (P40D680) at t = 60 minutes after s.c. injection, during which mice were awake and moving normally. K14-VEGFR-3-Fc mice showed signal at baseline levels (red dashed line) indicating no transport to blood. ***P < 0.001 (2-tailed Student’s t test). Data are the mean ± SD.

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