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Intravital imaging of intestinal lacteals unveils lipid drainage through contractility
Kibaek Choe, … , Gou Young Koh, Pilhan Kim
Kibaek Choe, … , Gou Young Koh, Pilhan Kim
Published October 5, 2015
Citation Information: J Clin Invest. 2015;125(11):4042-4052. https://doi.org/10.1172/JCI76509.
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Research Article Vascular biology

Intravital imaging of intestinal lacteals unveils lipid drainage through contractility

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Abstract

Lacteals are lymphatic vessels located at the center of each intestinal villus and provide essential transport routes for lipids and other lipophilic molecules. However, it is unclear how absorbed molecules are transported through the lacteal. Here, we used reporter mice that express GFP under the control of the lymphatic-specific promoter Prox1 and a custom-built confocal microscope and performed intravital real-time visualization of the absorption and transport dynamics of fluorescence-tagged fatty acids (FAs) and various exogenous molecules in the intestinal villi in vivo. These analyses clearly revealed transepithelial absorption of these molecules via enterocytes, diffusive distribution over the lamina propria, and subsequent transport through lacteals. Moreover, we observed active contraction of lacteals, which seemed to be directly involved in dietary lipid drainage. Our analysis revealed that the smooth muscles that surround each lacteal are responsible for contractile dynamics and that lacteal contraction is ultimately controlled by the autonomic nervous system. These results indicate that the lacteal is a unique organ-specific lymphatic system and does not merely serve as a passive conduit but as an active pump that transports lipids. Collectively, using this efficient imaging method, we uncovered drainage of absorbed molecules in small intestinal villus lacteals and the involvement of lacteal contractibility.

Authors

Kibaek Choe, Jeon Yeob Jang, Intae Park, Yeseul Kim, Soyeon Ahn, Dae-Young Park, Young-Kwon Hong, Kari Alitalo, Gou Young Koh, Pilhan Kim

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Figure 4

Contractile movement of lacteals.

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Contractile movement of lacteals.
(A) Representative image and (B) illus...
(A) Representative image and (B) illustration of a contracting lacteal. Asterisks indicate the same cell inside the lacteal. The white dotted line outlines the structure of the villus. The red dotted line indicates the point at which the lacteal diameter was measured. The red arrow indicates the positive direction used for cell velocity and villus motion measurement and analysis. Scale bar: 30 μm. (C) Images and their illustrations were taken at different time points to characterize the cell and villus movement. Scale bar: 30 μm. (D) Illustrations of the tip of the lacteal to represent lacteal contraction. The red dotted lines outline the structure of lacteal at the previous time point. (E) Comparison of lacteal diameter and cell velocity versus time. (F) Comparison of lacteal diameter and villus displacement versus time. (G) Comparison of cell velocity and villus velocity versus time. (H) Comparison of the clearing rate of C12 or C16 FAs, which was quantified by calculating and comparing the change of average fluorescence intensity in the lamina propria of lowly contracting lacteals against highly contracting lacteals. Low and high contraction were defined as the average of two measurements of lacteal contraction being either lower or higher than 0.81 μm•Hz, respectively. A total of 36 villi from 4 mice and 36 villi from 3 mice were analyzed for C12 and C16 FAs, respectively. *P < 0.05, paired Student’s t test. Error bars indicate SEM.

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ISSN: 0021-9738 (print), 1558-8238 (online)

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