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Journal of Lipid Research, Vol. 50, 1080-1089, June 2009
Copyright © 2009 by American Society for Biochemistry and Molecular Biology

A dynamic, cytoplasmic triacylglycerol pool in enterocytes revealed by ex vivo and in vivo coherent anti-Stokes Raman scattering imaging

Jiabin Zhu^*, Bonggi Lee, Kimberly K. Buhman^1, and Ji-Xin Cheng^1,*,

^* Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907
Department of Foods and Nutrition, Purdue University, West Lafayette, IN 47907
Department of Chemistry, Purdue University, West Lafayette, IN 47907

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of eight figures.

This work was partially supported by Purdue Research Foundation grants to J-X.C. and K.K.B. and institutional support to K.K.B.

Published, JLR Papers in Press, February 18, 2009.

¹ To whom correspondence should be addressed. e-mail: kbuhman{at}purdue.edu (K.K.B.); jcheng{at}purdue.edu (J-X.C.)

The absorptive cells of the small intestine, enterocytes, are not generally thought of as a cell type that stores triacylglycerols (TGs) in cytoplasmic lipid droplets (LDs). We revisit TG metabolism in enterocytes by ex vivo and in vivo coherent anti-Stokes Raman scattering (CARS) imaging of small intestine of mice during dietary fat absorption (DFA). We directly visualized the presence of LDs in enterocytes. We determined lipid amount and quantified LD number and size as a function of intestinal location and time post-lipid challenge via gavage feeding. The LDs were confirmed to be primarily TG by biochemical analysis. Combined CARS and fluorescence imaging indicated that the large LDs were located in the cytoplasm, associated with the tail-interacting protein of 47 kDa. Furthermore, in vivo CARS imaging showed real-time variation in the amount of TG stored in LDs through the process of DFA. Our results highlight a dynamic, cytoplasmic TG pool in enterocytes that may play previously unexpected roles in processes, such as regulating postprandial blood TG concentrations.

Supplementary key words small intestine • dietary fat absorption • CARS microscopy

Abbreviations: CARS, coherent anti-Stokes Raman scattering; CLD, cytoplasmic lipid droplet; CM, chylomicron; DFA, dietary fat absorption; DGAT1, diacylglycerol acyltransferase 1; E-CARS, epidetected coherent anti-Stokes Raman scattering; ER, endoplasmic reticulum; F-CARS, forward-detected coherent anti-Stokes Raman scattering; LD, lipid droplet; MG, monoacylglycerol; NA, numerical aperture; ORO, oil red O; PDI, protein disulfide isomerase; TG, triacylglycerol; TIP47, tail-interacting protein of 47 kDa; TPEF, two-photon excited fluorescence

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