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Lee, Woo, Kang, Eom, Kang, Hwang, Kim, and Song: Meibography and Immunohistochemistric Structures in Animal Models
Original Article

Journal of the Korean Ophthalmological Society 2018; 59(1): 9-16.

Published online: 12 January 2018

DOI: https://doi.org/10.3341/jkos.2018.59.1.9

Meibography and Immunohistochemistric Structures in Animal Models

Hyun Kyu Lee, MD1, In Ho Woo, MD1, Boram Kang, MS1, Young Sub Eom, MD, PhD1, Su-Yeon Kang, MD, PhD1, Ho Sik Hwang, MD, PhD2, Hyo Myung Kim, MD, PhD1, Jong Suk Song, MD, PhD1

1Department of Ophthalmology, Korea University College of Medicine, Seoul, Korea.

2Department of Ophthalmology, Chuncheon Sacred Heart Hospital, Hallym University College of Medicine, Chuncheon, Korea.

Address reprint requests to Jong Suk Song, MD, PhD. Department of Ophthalmology, Korea University Guro Hospital, #148 Gurodong-ro, Guro-gu, Seoul 08308, Korea. Tel: 82-2-2626-1277, Fax: 82-2-857-8580, crisim@korea.ac.kr

Received 14 September 2017       Revised 30 October 2017       Accepted 19 December 2017

©2018 The Korean Ophthalmological Society

The Korean Ophthalmological Society

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

We evaluated differences in the meibographic findings and immunohistochemical structures of mice, rats, and rabbits (commonly used in animal experiments).

Methods

Meibomian gland images were taken using an animal meibograph in mice, rats, and black and white rabbits (five of each animal).The size ratios of the upper and lower Meibomian glands were calculated. Afterm meibographic imaging, the eyes were cut sagittally or transversely and the histological structures were compared using hematoxylin-and-eosin (H&E) and immunohistochemical staining. In addition, the diameters of the central ducts and the areas of ducted glands and the acini were compared using image analysis software.

Results

The meibographic findings were clearer in black rabbits than in the other animals. The upper to lower length ratio of the gland did not differ significantly among the three species. Histologically, no central duct epithelial cells were apparent in mice. The diameters of the central ducts and the areas of ducted glands and acini increased in the following order: mouse, rat, and rabbit. However, the Meibomian gland areas did not differ significantly between mice and rats. On immunohistochemical staining, the central ductal epithelium and the Meibomian gland acini were stained for cytokeratin 5 (CK 5) but only the central ductal epithelium was stained for cytokeratin 6 (CK 6).

Conclusions

The larger the experimental animal, the greater the sizes of the Meibomian gland ducts and acini. In addition, black rabbits yielded better gland images because contrast was enhanced. Compared with sagittal sections of Meibomian glands, transverse sections facilitated a better understanding of the structures of the central ducts and surrounding Meibomian gland acini.

Keywords: Animal, Meibography, Meibomian gland, Immunohistochemistry

Figures and Tables

Figure 1

Meibography for animals. (A) Meibography for animals. (B) The animals were anesthetized by intramuscular injection, and meibography of animals was performed.

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Figure 2

Meibographic images of upper and lower eyelids using Meiboviewer for animal experiments. Black mice (A), white mice (B), white rat (C), black rabbit (D), and white rabbit (E).

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Figure 3

Histologic results of meibomian gland. The structures such as the central duct and the surrounding lobules of the rabbit were observed more uniformly than the other animals. In rats and rabbits, ductal epithelial cells were observed, but in the mouse, epithelial cells of the central duct were not clearly observed (Upper: mouse, Middle: rat, Lower: rabbit) (Hematoxylin and Eosin [H&E] stain, A: ×40, B: ×200).

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Figure 4

Immunohistochemistry results of meibomian gland (CK 5, CK 6). In the case of mouse, the size of the lobules and the central ducts was small, so that the difference between CK 5 and CK 6 was not apparent (A). In addition, it was difficult to determine whether the central duct epithelial cells were stained in both CK 5 and CK 6 (A). However, in rats, CK 5 was stained in the epithelium and lobules of the duct, and CK 6 was stained only in the ductal epithelial cells (B). In rabbits, the difference between CK 5 and CK 6 was most obvious (C) (A: Mouse, B: Rat, C: Rabbit) (Left: CK 5, Right: CK 6, ×100).

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Figure 5

Structures of meibomian glands in different section. In sagittal section (A), although there is an advantage of observing the entire meibomian gland, there is a disadvantage in that it cannot confirm the structure of the gland structure because there is difference in the structure depending on the cutting position. On the other hand, in Transverse section (B), only the portion corresponding to the cutting position can be observed, but the relationship between the central tube and the surrounding lobules could be seen more clearly (A: Sagittal section, B: Transverse section, Hematoxylin and Eosin [H&E] stain, ×40).

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Table 1

Animal models (mouse, rat, rabbit)

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Table 2

Ratio of upper meibomian gland length to lower meibomian gland length in mouse, rat and rabbit

jkos-59-9-i002

Values are presented as mean ± SD unless otherwise indicated.

Table 3

Central duct, acinar area and central duct diameter in mouse, rat and rabbit

jkos-59-9-i003

Values are presented as mean ± SD unless otherwise indicated.

*Analysis of variance (ANOVA).

Notes

Conflicts of Interest The authors have no conflicts to disclose.

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