Presence of adipose differentiation-related protein in rat meibomian gland cells.
Adipose differentiation-related protein (ADRP) is an intrinsic lipid storage protein found in lipid droplets of different type of cells. ADRP has been recognized to be a specific marker of lipid accumulation and a marker of differentiated adipocytes. The purpose of this study was to determine whether ADRP was present in the cells of the meibomian gland. The expression of the mRNA of ADRP was determined by RT-PCR and Northern blot analysis of the meibomian gland and other rat tissues. A newly generated polyclonal antibody against rat ADRP was used for Western blot analysis and immunohistochemical staining to determine whether ADRP was expressed in the rat meibomian gland. Meibomian gland acinar cells were isolated to determine when ADRP was expressed during cell differentiation in vitro. Northern blot analysis and Western analysis showed that ADRP was expressed in the meibomian gland. Immunoreactivity to ADRP was observed in the lobules of acinar cells in the meibomian gland, and was preferentially located adjacent the vacuolated cytoplasm. In culture, the meibocytes began to store lipid droplets in the cytoplasm as they became confluent, and the immunoreactivity for ADRP was found at the margins of the oil droplets. Our results suggest that ADRP can serve as a new marker for the identification of differentiated meibocytes containing lipid droplets.
Kutsuna
Miki
M
Department of Ophthalmology, School of Medicine, Ehime University, Shitsukawa, Toon-City, Ehime 791-0295, Japan. mittan@m.ehime-u.ac.jp
Kodama
Toshio
T
Sumida
Michihiro
M
Nagai
Ayako
A
Higashine
Masakatsu
M
Zhang
Wei
W
Hayashi
Yasuhito
Y
Shiraishi
Atsushi
A
Ohashi
Yuichi
Y
D000273:Adipose Tissue / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D005136:Eye Proteins / Q000032:analysis* / Q000276:immunology; D007150:Immunohistochemistry / Q000379:methods; D008055:Lipids / Q000032:analysis; D008297:Male; D008537:Meibomian Glands / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D000070780:Perilipin-2; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D005136:Eye Proteins; D008055:Lipids; D008565:Membrane Proteins; D000070780:Perilipin-2; C000602984:Plin2 protein, rat; D012333:RNA, Messenger