This article is part of the supplement: 11th International Symposium on the Cells of the Hepatic Sinusoid and their Relation to Other Cells .

Proceedings

Intralobular Distribution of Vitamin A-Storing Lipid Droplets in Hepatic Stellate Cells with Special Reference to Polar Bear and Arctic Fox

Nobuyo Higashi¹ , Katsuyuki Imai¹ , Mitsuru Sato¹ , Takeya Sato¹ , Naosuke Kojima¹ , Mitsutaka Miura¹ , Heidi L Wold³ , Jan Øivind Moskaug³ , Trond Berg⁴ , Kaare R Norum³ , Norbert Roos⁵ , Kenjiro Wake² , Rune Blomhoff³ and Haruki Senoo¹

¹  Department of Anatomy, Akita University School of Medicine, Akita 010-8543, Japan

²  Liver Research Unit, Minophagen Pharmaceutical Co. Ltd., Tokyo 160-0004, Japan

³  Institute for Nutrition Research, Faculty of Medicine, University of Oslo, Norway

⁴  Division of Molecular Biology, Institute of Biology, University of Oslo, Norway

⁵  Electronmicroscopical Unit for Biological Sciences, Faculty of Mathematics and Natural Sciences, University of Oslo, Norway

author email corresponding author email

Comparative Hepatology 2004, 3(Suppl 1):S16doi:10.1186/1476-5926-2-S1-S16

The electronic version of this article is the complete one and can be found online at: http://www.comparative-hepatology.com/content/3/S1/S16

Published:

14 January 2004

© 2004 Higashi et al; licensee BioMed Central Ltd
This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

We examined the liver of adult polar bears, arctic foxes, and rats by gold chloride staining, fluorescence microscopy for the detection of autofluorescence of vitamin A, hematoxylin-eosin staining, staining with Masson's trichrome, Ishii and Ishii's silver impregnation, and transmission electron microscopical morphometry. The liver lobules of the arctic animals showed a zonal gradient in the storage of vitamin A. The density (i.e., cell number per area) of hepatic stellate cells was essentially the same among the zones. These results indicate that the hepatic stellate cells of the polar bears and arctic foxes possess heterogeneity of vitamin A-storing capacity in their liver lobules.

Introduction

Hepatic stellate cells (vitamin A-storing cells, fat-storing cells, lipocytes, interstitial cells) are located in the perisinusoidal space of Disse and extend their thin fibrillar processes into this space [1,2]. Under physiological conditions, the hepatic stellate cells can store 80% of the total vitamin A in the whole body as retinyl esters in lipid droplets in the cytoplasm and play pivotal roles in regulation of vitamin A homeostasis [3]. We have reported that arctic animals such as polar bear store a large amount of vitamin A in hepatic stellate cells as compared to human or usual experimental animals such as rats or mice. However, the question as to the distribution of vitamin A-storing lipid droplets in hepatic stellate cells within the liver lobule remains unsettled. Therefore, we conducted the present study on polar bears and arctic foxes.

Methods

After having obtained permission to hunt animals from the District Governor of Svalbard, we caught 11 arctic foxes (Alopex lagopus) during the period from August 1996 to September 2001. Three polar bears (Ursus maritimus) were shot in self-defense in February and August 1998. We examined the liver of these animals by gold chloride staining, fluorescence microscopy for the detection of autofluorescence of vitamin A, hematoxylin-eosin staining, staining with Masson's trichrome, Ishii and Ishii's silver impregnation, and transmission electron microscopical morphometry using a division map of liver lobule for a zonal analysis (Figure 1). As a control, we examined the liver of rats in the same procedure.

Results

The liver lobules of the arctic animals showed a zonal gradient in the storage of vitamin A (Figure 2). The gradient was expressed as a symmetric crescendo-decrescendo profile starting at periportal zone, peaking at the middle zone, and sloping down toward the central zone in the liver lobule. The area of lipid droplets in the middle zone of polar bears was the largest in all zones evaluated. It was about 3 times as large as that of middle zone in arctic foxes, and about 43 times as large as that of the corresponding zone in rats. We also compared the cell density of stellate cells in each zone, but the cell density in each zone showed no significant differences (data not shown). The zonal differences revealed by light microscopical methods (data not shown) were consistent with electron microscopic morphometry. No pathological signs such as liver cirrhosis or hepatic fibrosis were observed in these animals (data not shown).

Discussion

Results of this study indicate that arctic animals possess the intralobular heterogeneity of vitamin A-storage capacity in the hepatic stellate cells.

In earlier studies, a specific zonality for intralobular vitamin A-storage was reported. Although our results agree with other reports as to the presence of the heterogeneity of vitamin A-storing lipid droplets in the hepatic stellate cells in the liver lobule [4,5], the location of the highest-storage zone disagree with these reports. This discrepancy might be explained by the observation methods used in the earlier studies. Whereas those authors used only the light microscopical method [4,5], in the present study we examined all tissues by electron microscopy. Moreover, we certified that the tendency of vitamin A storage observed in liver lobules by light microscopy strongly supported that of intralobular heterogeneity revealed by electron microscopical morphometry.

The existence of the intralobular heterogeneity of vitamin A storage would relate to the number of hepatic stellate cells within the each zone or the maturation of hepatic stellate cells itself [6]. However, regarding the quantification of cell density in each zone, the average density calculated in all the animal species examined in our study did not differ among the zones. In previous papers we reported that hepatic stellate cells displayed a different response to extracellular matrix (ECM) components to change their shape and cellular functions [7]. In addition, the heterogeneity of ECM in the liver lobule [8] and the modulation of the cellular retinol-binding protein (CRBP) level by the extracellular collagen matrix in the hepatic stellate cells have been reported [9]. CRBP plays an important role in retinol metabolism and has also been reported to be indispensable for efficient retinyl ester synthesis and storage [10]. Furthermore, Kato et al. [11] has reported the existence of the intralobular heterogeneity of CRBP in the liver of rats.

Hence, we speculate that the intralobular distribution of CRBP in the liver of polar bears and arctic foxes may also show similar heterogeneity to that in the rats, and have effects on the intralobular heterogeneity of storing vitamin A in the livers.

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