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Papers In Press, published online ahead of print January 1, 2007
J. Lipid Res., doi:10.1194/jlr.M600155-JLR200

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Journal of Lipid Research, Vol. 48, 9-18, January 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Temporal and spatial variations of lipid droplets during adipocyte division and differentiation

Masafumi Nagayama¹, Tsutomu Uchida and Kazutoshi Gohara

Division of Applied Physics, Graduate School of Engineering, Hokkaido University, North 13, West 8, Kita-ku, Sapporo 060-8628, Japan

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of 4 movies.

Published, JLR Papers in Press, October 21, 2006.

¹ To whom correspondence should be addressed. e-mail: macci{at}eng.hokudai.ac.jp

By capturing time-lapse images of primary stromal-vascular cells (SVCs) derived from rat mesenteric adipose tissue, we revealed temporal and spatial variations of lipid droplets (LDs) in individual SVCs during adipocyte differentiation. Numerous small LDs (a few micrometers in diameter) appeared in the perinuclear region at an early stage of differentiation; subsequently, several LDs grew to more than 10 µm in diameter and occupied the cytoplasm. We have developed a method for the fluorescence staining of LDs in living adipocytes. Time-lapse observation of the stained cells at higher magnification showed that nascent LDs (several 100 nm in diameter) grew into small LDs while moving from lamellipodia to the perinuclear region. We also found that adipocytes are capable of division and that they evenly distribute the LDs between two daughter cells. Immunofluorescence observations of LD-associated proteins revealed that such cell divisions of SVCs occurred even after LDs were coated with perilipin, suggesting that the "final" cell division during adipocyte differentiation occurs considerably later than that characterized in 3T3-L1 cells. Our time-lapse observations have provided a detailed account of the morphological changes that SVCs undergo during adipocyte division and differentiation.

Supplementary key words time-lapse observation • fluorescence observation • live cell imaging • visceral adipocyte • adipogenesis • adipose conversion • lipid bodies • lipid droplet-associated proteins

Abbreviations: ADRP, adipose differentiation-related protein; DIC, differential interference contrast; LD, lipid droplet; PhC, phase-contrast; PPAR-, peroxisome proliferator-activated receptor-; SVC, stromal-vascular cell

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