Minimally invasive quantification of lymph flow in mice and rats by imaging depot clearance of near-infrared albumin.
Identifieur interne : 004127 ( Main/Merge ); précédent : 004126; suivant : 004128Minimally invasive quantification of lymph flow in mice and rats by imaging depot clearance of near-infrared albumin.
Auteurs : Tine V. Karlsen [Norvège] ; Emmet Mccormack ; Maja Mujic ; Olav Tenstad ; Helge WiigSource :
- American journal of physiology. Heart and circulatory physiology [ 1522-1539 ] ; 2012.
Descripteurs français
- KwdFr :
- MESH :
- métabolisme : Albumines.
- physiologie : Lymphe, Système lymphatique.
- Animaux, Colorants fluorescents, Femelle, Radiopharmaceutiques, Rats, Souris.
English descriptors
- KwdEn :
- MESH :
- chemical , metabolism : Albumins.
- physiology : Lymph, Lymphatic System.
- Animals, Female, Fluorescent Dyes, Mice, Radiopharmaceuticals, Rats.
Abstract
There is a lack of available methods to noninvasively quantify lymphatic function in small experimental animals, a necessity for studies on lymphatic system pathophysiology. We present a new method to quantify lymph flow in mice and rats, based on optically monitoring the depot clearance of near-infrared fluorescently labeled albumin and subsequent calculation of removal rate constants (k). BSA was conjugated with Alexa680 NHS ester and remained stable in protein-rich solutions without free dye dissociation. To assess lymph flow, mice or rats were imaged every 30 or 60 min during a 3- to 6-h period following an intradermal injection of 0.5 or 1 μl Alexa680-albumin. Mice were awake between measurements, whereas rats were anesthetized throughout the experiment. The k, a parameter defined as equivalent to lymph flow, was calculated from the slopes of the resultant log-linear washout curves and averaged -0.40 ± 0.03 and -0.30 ± 0.02%/min for control C57BL/6 and C3H mice, respectively. Local administration of the vasoconstrictor endothelin-1 in mice led to a significant reduction in k, whereas overhydration in rats increased k, reflecting the coupling between capillary filtration and lymph flow. Furthermore, k was 50% of wild type in lymphedema Chy mice where dermal lymphatics are absent. We conclude that lymph flow can be determined as its rate constant k by optical imaging of depot clearance of submicroliter amounts of Alexa680-albumin. The method offers a minimally invasive, reproducible, and simple alternative to assess lymphatic function in mice and rats.
DOI: 10.1152/ajpheart.00842.2011
PubMed: 22101523
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pubmed:22101523Le document en format XML
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<term>Radiopharmaceutiques</term>
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<front><div type="abstract" xml:lang="en">There is a lack of available methods to noninvasively quantify lymphatic function in small experimental animals, a necessity for studies on lymphatic system pathophysiology. We present a new method to quantify lymph flow in mice and rats, based on optically monitoring the depot clearance of near-infrared fluorescently labeled albumin and subsequent calculation of removal rate constants (k). BSA was conjugated with Alexa680 NHS ester and remained stable in protein-rich solutions without free dye dissociation. To assess lymph flow, mice or rats were imaged every 30 or 60 min during a 3- to 6-h period following an intradermal injection of 0.5 or 1 μl Alexa680-albumin. Mice were awake between measurements, whereas rats were anesthetized throughout the experiment. The k, a parameter defined as equivalent to lymph flow, was calculated from the slopes of the resultant log-linear washout curves and averaged -0.40 ± 0.03 and -0.30 ± 0.02%/min for control C57BL/6 and C3H mice, respectively. Local administration of the vasoconstrictor endothelin-1 in mice led to a significant reduction in k, whereas overhydration in rats increased k, reflecting the coupling between capillary filtration and lymph flow. Furthermore, k was 50% of wild type in lymphedema Chy mice where dermal lymphatics are absent. We conclude that lymph flow can be determined as its rate constant k by optical imaging of depot clearance of submicroliter amounts of Alexa680-albumin. The method offers a minimally invasive, reproducible, and simple alternative to assess lymphatic function in mice and rats.</div>
</front>
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